Objective:The fruit stalk of Schisandra chinensis(Turcz.)Baill.(S.chinensis)has been found to contain bioactive components similar to the fruit of S.chinensis.Here,we report a recent discovery about new nortriterpenoi...Objective:The fruit stalk of Schisandra chinensis(Turcz.)Baill.(S.chinensis)has been found to contain bioactive components similar to the fruit of S.chinensis.Here,we report a recent discovery about new nortriterpenoids with a novel skeleton and anti-gastric cancer activity,which were isolated from the fruit stalk of S.chinensis.Methods:The chemical components of ethyl acetate extract from 70%ethanol extract from S.chinensis fruit stalk were separated,purified,and identified by liquid chromatography methods(silica gel,ODS,HPLC)and extensive spectroscopic analyses(NMR,IR,UV,MS,CD).Results:Two new nortriterpenoids,schilancitrilactone M and 25-hydroxyl schindilactone D(1 and 2),along with ten known nortriterpenoids(3-12)were isolated from the fruit stalk of S.chinensis.The isolated compounds were tested for their cytotoxic activities against MGC-803 cells,and the results showed that compounds 6-8 possessed significant activities with IC50 of 9.01,11.77,and 2.74μmol/L,respectively.Conclusion:Twelve nortriterpenoids including two new compounds were isolated from the fruit stalk of S.chinensis for the first time.Among them,compounds 6-8 showed significant anti-gastric cancer activities.We postulated that the fruit stalk of S.chinensis could be used as an anti-gastric cancer drug.展开更多
Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF ...Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF has anti-inflammatory,protective liver,antioxidant,antibacterial and other pharmacological effects.Ancient prescriptions are commonly used in the treatment of chronic diarrhea and other intestinal diseases and diabetes.Modern clinical pharmacology features of SCF polysaccharide(SCFP)in diabetes,liver diseases,enteritis and other aspects have achieved excellent results.Gut is an important digestive organ of human body,but intestinal diseases are varied,including Crohn′s disease,ulcerative colitis,intestinal flora imbalance,etc..It is a chronic and non-specific inflammatory disease.The disease is persisted for a long time and the incidence rate is expected to rise.Most of the symptoms are recurrent diarrhea,bloody stool and abdominal pain.It is considered by the World Health Organization as a refractory disease.At present,there is little possibility of complete cure,which is closely related to complex environmental factors,eating habits and heredity.In recent years,clinical studies have found that SCFP has a variety of pharmacological effects on intestinal protection.①Reduce inflammatory factors:intestinal mucositis is a common adverse reaction in patients with chemotherapy.The development of mucositis is related to pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL^(-1)β,Interferon-γ(IFN-γ).SCFP can significantly reduce IL-6 TNF-α,IL^(-1)β,and IL-8,as well as the accumulation of T cells in the process of resisting apoptosis,reduce the inflammatory reaction and protect the damage to villi and crypts,improve the symptoms of small intestinal mucositis caused by weight loss and diarrhea.②Promote immunoglobulin A secretion:intestinal mucosal immunity is the first line of defense of the body′s immune system.Its main antibody is secretory immunoglobulin A,which can destroy and phagocytize microorganisms,bacteria and viruses.SCFP can improve intestinal immunity by increasing the number and activity of T lymphocytes,promoting the secretion of secretory immunoglobulin A,and affecting the activity of a variety of cytokines.③Regulation of intestinal flora:the flora in the intestine has the functions of auxiliary nutrient absorption,biological antagonism and immune regulation,and can form a natural barrier for the host's intestine.When the human intestinal flora is disordered,probiotics will be greatly reduced,harmful bacteria will proliferate and destroy the intestinal environment.Under these conditions,the intake of SCFP significantly increased the number of beneficial bacteria such as bifidobacteria and lactobacillus,and significantly decreased the number of conditional pathogens such as enterococcus and escherichia coli,indicating that SCFP can indeed regulate the intestinal disorder caused by lincomycin hydrochloride to a certain extent. This may be because beneficial bacteria in the intestine metabolize polysaccharides produce short chain fatty acids such as lactic acid and acetic acid, which reduces the pH value in the intestine and inhibits the growth of enterococcus and Escherichia coli. In conclusion, SCFP can treat and protect intestinal diseases to a certain extent, which provides a favorable basis for the treatment of intestinal diseases.展开更多
Schisandra chinensis (Turcz.) Baill. is a deciduous woody vine, and belongs to the Schisandraceae family. This species is a horticultural plant with edible fruit. It is used medicinally and for making rope. It is dist...Schisandra chinensis (Turcz.) Baill. is a deciduous woody vine, and belongs to the Schisandraceae family. This species is a horticultural plant with edible fruit. It is used medicinally and for making rope. It is distributed in Hebei, Heilongjiang, Jilin, Liaoning, Nei Mongol,展开更多
BACKGROUND:The most prominent characteristic of brain aging is decreased learning and memory ability.The functions of learning and memory are closely related to intracerebral acetylcholinesterase(AChE) and monoamine n...BACKGROUND:The most prominent characteristic of brain aging is decreased learning and memory ability.The functions of learning and memory are closely related to intracerebral acetylcholinesterase(AChE) and monoamine neurotransmitter activity.Previous studies have shown that Schisandra chinensis polysaccharide has an anti-aging effect. OBJECTIVE:To explore the effects of Schisandra chinensis polysaccharide on AChE activity and monoamine neurotransmitter content,as well as learning and memory ability in a D-galactose-induced aging mouse brain model compared with the positive control drug Kangnaoling. DESIGN,TIME AND SETTING:Completely randomized,controlled experiment based on neurobiochemistry was performed at the Pharmacological Laboratory,Henan University of Traditional Chinese Medicine from September to December 2003. MATERIALS:Schisandra chinensis was purchased from Henan Provincial Medicinal Company. Schisandra chinensis polysaccharide was obtained by water extraction and alcohol precipitation. Kangnaoling pellets were provided by Liaoning Tianlong Pharmaceutical(batch No.20030804;state drug permit No.H21023095).A total of 50 six-week-old Kunming mice were randomly divided into five groups:blank control,model,Kangnaoling,high and low dosage Schisandra chinensis polysaccharide groups,with 10 mice per group. METHODS:Mice in the blank control group were subcutaneously injected with 0.5 mL/20 g normal saline into the nape of the neck each day,while the remaining mice were subcutaneously injected with 5%D-galactose saline solution(0.5 mL/20 g) in the nape for 40 days to induce a brain aging model.On day 11,mice in the high and low dosage Schisandra chinensis polysaccharide groups were intragastrically infused with 20 mg/mL and 10 mg/mL Schisandra chinensis polysaccharide solution(0.2 mL/10 g),respectively.Mice from the Kangnaoling group were intragastrically infused with 35 mg/mL Kangnaoling suspension(0.2 ml/10 g),and the mice in the model group were intragastrically infused with the same volume of normal saline(0.2 mL/10 g) once per day for 30 consecutive days. MAIN OUTCOME MEASURES:Two hours after the final administration,pathohistological changes in the cerebral cortex and hippocampus were observed using hematoxylin & eosin staining.AChE activity was detected using chromatometry.Monoamine neurotransmitter content was measured using fluorimetry.Learning and memory was measured using the step down test and darkness avoidance test. RESULTS:Both Schisandra chinensis polysaccharide and Kangnaoling improved pathological injury to the cerebral cortex and hippocampus in a mouse model of brain aging.Compared with the blank control group,AChE activity and content of norepinephrine(NA),dopamine(DA),and 5-hydroxytryptamine(5-HT) were significantly decreased in the model group(P<0.01).In contrast, AChE activity and NA,DA,and 5-HT levels significantly increased in the Kangnaoling and high dosage Schisandra chinensis polysaccharide groups(P<0.01),while NA levels significantly increased in the low dosage Schisandra chinensis polysaccharide group(P<0.01).Drug treatment improved learning and memory abilities(P<0.01 or P<0.05). CONCLUSION:Schisandra chinensis polysaccharide significantly increased levels of central neurotransmitters and improved learning and memory in a mouse model of brain aging.The effects of Schisandra chinensis polysaccharide were equal to that of Kangnaoling pellets.展开更多
The essential oil of Schisandra Chinensis(Turc z.) Baill was extracted and analyzed. The essential oil was obtained in a 1.8%(mL/100 g material) yield. Forty eight compounds were isolated and characterized from the oi...The essential oil of Schisandra Chinensis(Turc z.) Baill was extracted and analyzed. The essential oil was obtained in a 1.8%(mL/100 g material) yield. Forty eight compounds were isolated and characterized from the oil. The results show that the content(20.42%) of β -pinene of naturally essential oil is the highest among the components of the oil,which has laid a foundation for the futher development and application of the stems of Schisandra Chinensis(Turc z.) Baill .展开更多
The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy(HPLC).The sample was extracted exceedingly in the critical limit of CO 2 ....The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy(HPLC).The sample was extracted exceedingly in the critical limit of CO 2 .The retention time of Schisandrin A and Schisandrin B was reduced,with methano/water(75:25)as mobile phase.The wavelength for detection was 254 nm. The R2 of standard curve was 0.9998 and the relative standard deviation was 2.31%and 3.17%with the recovery of 96.45%and 97.37%,respectively.The result shows that the rate of veracity of this method is higher and it proves that the determination method of Schisandrin A and Schisandrin B in Schisandra chinensis is a feasible method.展开更多
OBJECTIVE To study the pharmacokinetics change of schisandra chinensis under the pathological condition of liver dysfunction for safe and rational use of herbal medicines.METHODS The metabolism of four effective ligna...OBJECTIVE To study the pharmacokinetics change of schisandra chinensis under the pathological condition of liver dysfunction for safe and rational use of herbal medicines.METHODS The metabolism of four effective lignans from schisandra chinensis(SC),schisandrin,schisantherin A,deoxyshisandrin and γ-schisandrin was studied using microsomes from patients with advanced hepato.cellular carcinoma.In situ intestinal and hepatic perfusions were conducted to clarify the contributions from impairments of gut and liver on the pharmacokinetics of the four schisandra lignans in CCl4-intoxi.cated rats.The metabolism in rat and human liver microsomes and transport in Caco-2 monolayer cell model were studied to reveal the key factors for the in vivo disposition of the four lignans.RESULTS When SC alcoholic extract was orally administrated to CCl4-intoxicated rat for a short term(4 d),the pharmacokinetics of four active SC lignans was significantly changed while its hepatoprotective effect was not obviously observed.The plasma concentrations of the four schisandra lignans were dramatical.ly elevated compared with the control.The Cmax,AUC and MRT were all increased or prolonged signif.icantly while parameter CLz/F was obviously reduced in rat pretreated with CCl4.In hepatic perfusion study and liver microsomes incubation,it was found that the hepatic metabolism of the four lignans was markedly decreased mainly due to the activity reduction of multiple CYP450 isoenzymes involved the metabolism,which,eventually,might lead to the alternation of their pharmacokinetic profiles in CCl4-intoxicated rats or patients with advanced hepatocellular carcinoma.CONCLUSION The pharmacoki.netic studies of SC components in pathological situation of liver dysfunction are expected to provide useful data for rational and safe application of SC preparations in clinic or further pharmacological and toxicological research.展开更多
[Objectives]To study the quality standard of Chinese drug in compound Qianglinaogingsu Tablets(Fructus Schisandrae Chinensis and Coru Cervi Pantotrichum).[Methods]The thin layer chromatography(TLC)differentiation meth...[Objectives]To study the quality standard of Chinese drug in compound Qianglinaogingsu Tablets(Fructus Schisandrae Chinensis and Coru Cervi Pantotrichum).[Methods]The thin layer chromatography(TLC)differentiation method was established for Fructus Schisandrae Chinensis and Cornu Cervi Pantotrichum.[Results]TLC spots developed were fairly clear,and the blank test showed no interference.[Conclusions]The method is fast,simple and can be used for quality control of Qianglinaoqingsu Tablets.展开更多
As highly social animals,Indo-Pacific humpback dolphins(Sousa chinensis)exhibit community differentiation.Nevertheless,our understanding of the external and internal factors influencing these dynamics,as well as their...As highly social animals,Indo-Pacific humpback dolphins(Sousa chinensis)exhibit community differentiation.Nevertheless,our understanding of the external and internal factors influencing these dynamics,as well as their spatiotemporal variations,is still limited.In the present study,variations in the social structure of an endangered Indo-Pacific humpback dolphin population in Xiamen Bay,China,were monitored over two distinct periods(2007–2010 and 2017–2019)to analyze the effects of habitat utilization and the composition of individuals within the population.In both periods,the population demonstrated a strikingly similar pattern of social differentiation,characterized by the division of individuals into two main clusters and one small cluster.Spatially,the two primary clusters occupied the eastern and western waters,respectively,although the core distribution area of the eastern cluster shifted further eastward between the two periods.Despite this distribution shift,the temporal stability of the social structure and inter-associations within the eastern cluster remained unaffected.A subset of 16individuals observed in both periods,comprising 51.6%and 43.2%of the population in each respective period,emerged as a foundational element of the social structure and may be responsible for sustaining social structure stability,especially during the 2007–2010 period.These observations suggest that the composition of dominant individuals,an internal factor,had a more substantial influence on the formation of the social network than changes in habitat use,an external factor.Consequently,the study proposes distinct conservation measures tailored to each of the two main clusters.展开更多
Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by usin...Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by using syntenic analysis combined with phylogenomic analysis of C.chinensis and four other representative genomes from basal and core eudicots,we confirmed that the WGD event in C.chinensis was shared by Aquilegia coerulea and Papaver somniferum L.and quickly occurred after Ranunculales diverged from other eudicots,likely a Ranunculales common tetraploidization(RCT).The synonymous nucleotide substitutions at synonymous sites distribution of syntenic blocks across these genomes showed that the evolutionary rate of the P.somniferum genome is faster than that of the C.chinensis genome by approximately 13.7%,possibly due to Papaveraceaes having an additional special tetraploidization event(PST).After Ks correction,the RCT dated to 115—130 million years ago(MYA),which was close to the divergence of Ranunculaceaes and Papaveraceaes approximately115.45—130.51 MYA.Moreover,we identified homologous genes related to polyploidization and speciation and constructed multiple sequence alignments with different reference genomes.Notably,the event-related subgenomes in the basal genomes all showed genomic fractionation bias,suggesting a likely allopolyploid nature of the RCT,PST and T-Alpha and T-Beta events in Tetracentron sinense.In addition,we detected that the sixteen P450 subfamilies were markedly expanded in the genomes of Ranunculales,and most of them were related to the RCT and PST events.We constructed a new platform for Early Eudicot Comparative Genomic Research(http://www.cgrpoee.top/index.html)to store more information.In summary,our findings support the WGD of C.chinensis shared by Ranunculales,which is likely an allotetraploidization event.This present effort offered new insights into the evolution of key polyploidization events and the genes related to secondary metabolites during the diversification of early eudicots.展开更多
The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.Th...The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.These similarities have established the tree shrew as a promising experimental model for biomedical research on cancer,infectious diseases,metabolic disorders,and mental health conditions.Herein,we used metatranscriptomic sequencing to analyze plasma,as well as oral and anal swab samples,from 105 healthy asymptomatic tree shrews to identify the presence of potential zoonotic viruses.In total,eight mammalian viruses with complete genomes were identified,belonging to six viral families,including Flaviviridae,Hepeviridae,Parvovirinae,Picornaviridae,Sedoreoviridae,and Spinareoviridae.Notably,the presence of rotavirus was recorded in tree shrews for the first time.Three viruses-hepacivirus 1,parvovirus,and picornavirus-exhibited low genetic similarity(<70%)with previously reported viruses at the whole-genome scale,indicating novelty.Conversely,three other viruses-hepacivirus 2,hepatovirus A and hepevirus-exhibited high similarity(>94%)to known viral strains.Phylogenetic analyses also revealed that the rotavirus and mammalian orthoreovirus identified in this study may be novel reassortants.These findings provide insights into the diverse viral spectrum present in captive Chinese tree shrews,highlighting the necessity for further research into their potential for crossspecies transmission.展开更多
The purpose of this project is used for exploring the mechanism of Callistephus chinensis in the treatment of diabetes by network pharmacology and molecular docking methods.The target of Callistephus chinensis was obt...The purpose of this project is used for exploring the mechanism of Callistephus chinensis in the treatment of diabetes by network pharmacology and molecular docking methods.The target of Callistephus chinensis was obtained from SwissTargetPrediction database,while the target related to diabetes was obtained from GeneCards and OMIM databases.The target was added in String database to build the protein interaction network.GO biological process enrichment analysis and KEGG pathway enrichment analysis were carried out by Metascape software,then the target-pathway network was constructed.Molecular docking was carried out in Discovery Studio 2016 Client software to verify the binding force of Callistephus chinensis flavonoid compounds with key targets.In this study,10 potential active components were selected from the flavonoid monomer compounds of Callistephus chinensis.1847 biological processes(BP),126 cell compositions(CC)and 256 molecular functions(MF)were obtained by GO enrichment analysis;a total of 194 pathways were involved in KEGG enrichment analysis of 192 cross targets.Network analysis showed that quercetin was the main active component of flavonoids in the treatment of diabetes,AKT1,TNF,VEGFA,EGFR,SRC and other related signals were in relation to the treatment of diabetes.This study showed that Callistephus chinensis flavonoid compounds play a role in the treatment of diabetes by regulating multi-target and multi-pathway.展开更多
Schisandra chinensis Turcz.(Baill.) is a plant species with fruits that have been well known in Far Eastern medicine for a long time. It has traditionally been used as a stimulating and fortifying agent in cases of ph...Schisandra chinensis Turcz.(Baill.) is a plant species with fruits that have been well known in Far Eastern medicine for a long time. It has traditionally been used as a stimulating and fortifying agent in cases of physical exhaustion and to inhibit fatigue.The major bioactive compounds found in S. chinensis are lignans with a dibenzocyclooctadiene skeleton, but little is known about their biosynthesis in plants. S. chinensis is the ideal medicinal plant for studying the biosynthesis of lignans, especially the dibenzocyclooctadiene skeleton. Genomic information for this important herbal plant is unavailable. To better understand the lignan biosynthesis pathway, we generated transcriptome sequences from the fruit during ripening and performed de novo sequence assembly, yielding136 843 unique transcripts with N50 of 1778 bp. Putative functions could be assigned to 41 824 transcripts(51.57%) based on BLAST searches against annotation databases including GO(Gene ontology) and KEGG(Kyoto Encyclopedia of Genes and Genomes). Furthermore, 22 candidate cytochrome P450 genes and 15 candidate dirigent proteins genes that were most likely involved in the lignan biosynthesis pathway were discovered based on transcriptome sequencing of S. chinensis. The genomic data obtained from S. chinensis, especially the identification of putative genes involved in the lignan biosynthesis pathway, will facilitate our understanding of lignan biosynthesis at the molecular level. The lignan metabolite profiles were analyzed by metabolomes, the accumulation patterns of 30 metabolites involved in the lignan pathway were studied. Co-expression network of lignan contents and transcriptional changes showed355 strong correlations(correlation coefficient, R^2 > 0.9) between 21 compounds and 153 transcripts. Furthermore, the comprehensive analysis and characterization of the genes involved in lignan pathways and the metabolite profiles of lignans are expected to provide better insight regarding the diversity of the chemical composition, synthetic characteristics, and regulatory mechanisms of this medical herb.展开更多
Objective To optimize the extract technology of active lignins from the fruits of Schisandra chinensis. Methods The content of schizandrin, gomisin A, and deoxyschizandrin were selected as standards to evaluate the ef...Objective To optimize the extract technology of active lignins from the fruits of Schisandra chinensis. Methods The content of schizandrin, gomisin A, and deoxyschizandrin were selected as standards to evaluate the efficiency of smashing tissue extraction (STE). Solid-liquid ratio, extracting times, ethanol concentration, and extracting time were investigated through orthogonal test. Results The optimized conditions for STE were ten times amount of 80% EtOH, extracting for three times, and 2 min for each time. Conclusion STE could obtain relatively higher yield, simplicity of operation, and benefit for environment protection. It could be better choice for the extraction of S. chinensis.展开更多
Six new lignans(1-6),as well as five known ones(7-11)were isolated from the leaves and stems of Schisandra chinensis.The structures of 1-6 were established on the basis of spectroscopic methods including 1D-and 2D-NMR...Six new lignans(1-6),as well as five known ones(7-11)were isolated from the leaves and stems of Schisandra chinensis.The structures of 1-6 were established on the basis of spectroscopic methods including 1D-and 2D-NMR techniques and CD experiments.Compound 1 was the first example of naturally occurring N-containing lignans featuring a nicotinoyl group.All the new compounds were evaluated for their anti-HIV-1 activities and showed EC50 values in the range 17.89-138.23μg/mL.展开更多
Objective: To investigate the protective effects of Schisandra chinensis oil(SCEO) against aristolochic acid Ⅰ(AAⅠ)-induced nephrotoxicity in vivo and in vitro and elucidate the underlying mechanism.Methods: C57BL/6...Objective: To investigate the protective effects of Schisandra chinensis oil(SCEO) against aristolochic acid Ⅰ(AAⅠ)-induced nephrotoxicity in vivo and in vitro and elucidate the underlying mechanism.Methods: C57BL/6 mice were randomly divided into 5 groups according to a random number table, including control group, AAⅠ group, and AAⅠ+SCEO(0.25, 0.5 and 1 g/kg) groups(n=5 per group). Pretreatment with SCEO was done for 2 days by oral administration, while the control and AAⅠ groups were treated with sodium carboxymethyl cellulose. Mice of all groups except for the control group were injected intraperitoneally with AAⅠ(5 mg/kg) from day 3 until day 7. Histopathological examination and apoptosis of kidney tissue were observed by hematoxylin and eosin and TdT-mediated dUTP nick-end labeling(TUNEL) staining, respectively. The levels of serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), blood urea nitrogen(BUN), and serum creatinine(SCr), as well as renal malondialdehyde(MDA), glutathione, r-glutamyl cysteingl+glycine(GSH), and superoxide dismutase(SOD) were analyzed using enzyme-linked immunosorbent assay(ELISA).Expressions of hepatic cytochrome P450 1A1(CYP1A1), CYP1A2, and nad(p)hquinonedehydrogenase1(NQO1) were analyzed using ELISA, quantitative real-time polymerase chain reaction(qPCR) and Western blot,respectively. In vitro, SCEO(40 μg/mL) was added 12 h before treatment with AAⅠ(40 μmol/mL for 48 h) in human renal proximal tubule cell line(HK-2), then apoptosis and reactive oxygen species(ROS) were analyzed by flow cytometry. Results: SCEO 0.5 and 1 g/kg ameliorated histopathological changes and TUNEL+ staining in the kidney tissues of mice with AAⅠ-induced nephrotoxicity, and reduced serum levels of ALT, AST, BUN and SCr(P<0.01 or P<0.05). SCEO 0.5 and 1 g/kg alleviated the ROS generation in kidney, containing MDA,GSH and SOD(P<0.01 or P<0.05). SCEO 1 g/kg increased the expressions of CYP1A1 and CYP1A2 and decreased NQO1 level in the liver tissues(P<0.01 or P<0.05). Besides, in vitro studies also demonstrated that SCEO 40 μg/mL inhibited apoptosis and ROS generation(P<0.05 or P<0.01). Conclusions: SCEO can alleviate AAⅠ-induced kidney damage both in vivo and in vitro. The protective mechanism may be closely related to the regulation of metabolic enzymes, thereby inhibiting apoptosis and ROS production.展开更多
Phenylalanine ammonia-lyase(PAL),which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid,is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropa...Phenylalanine ammonia-lyase(PAL),which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid,is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes.Schisandra chinensis,a woody vine plant belonging to the family of Magnoliaceae,is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity.However,the functional role of PAL in the biosynthesis of lignan is relatively limited,compared with those in lignin and flavonoids biosynthesis.Therefore,it is essential to clone and characterize the PAL genes from this valuable medicinal plant.In this study,molecular cloning and characterization of three PAL genes(ScPAL1−3)from S.chinensis was carried out.ScPALs were cloned using RACE PCR.The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis.In order to determine their catalytic activity,recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli(BL21-DE3),followed by Ni-affinity purification.The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds.The optimal temperature,pH value and effects of different metal ions were determined.Vmax,Kcat and Km values were determined under the optimal conditions.The expression of three ScPALs in different tissues was also determined.Our work provided essential information for the function of ScPALs.展开更多
The present study was designed to evaluate protective activity of an ethanol extract of the stems of Schisandra chinensis(SCE) and explore its possible molecular mechanisms on acetaminophen(APAP) induced hepatotoxicit...The present study was designed to evaluate protective activity of an ethanol extract of the stems of Schisandra chinensis(SCE) and explore its possible molecular mechanisms on acetaminophen(APAP) induced hepatotoxicity in a mouse model. The results of HPLC analysis showed that the main components of SCE included schisandrol A, schisandrol B, deoxyschisandrin, schisan--drin B, and schisandrin C and their contents were 5.83, 7.11, 2.13, 4.86, 0.42 mg·g^(-1), respectively. SCE extract was given for 7 con--secutive days before a single hepatotoxic dose of APAP(250 mg·kg^(-1)) was injected to mice. Our results showed that SCE pretreatment ameliorated liver dysfunction and oxidative stress, which was evidenced by significant decreases in aspartate transaminase(AST), alanine aminotransferase(ALT), malondialdehyde(MDA) contents and elevations in reduced glutathione(GSH) and superoxide dismutase(SOD) levels. These findings were associated with the result that the SCE pretreatment significantly decreased expression levels of 4-hydroxynonenal(4-HNE) and 3-nitrotyrosine(3-NT). SCE also significantly decreased the expression levels of Bax, mitogen-activated protein kinase(MAPK), and cleaved caspase-3 by APAP exposure. Furthermore, supplementation with SCE suppressed the expression levels of inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2), suggesting alleviation of inflammatory response. In summary, these findings from the present study clearly demonstrated that SCE exerted significant alleviation in APAP-induced oxidative stress, inflammation and apoptosis mainly via regulating MAPK and caspase-3 signaling pathways.展开更多
基金This study was supported by the National Nature Science Foundation(81973440)National Key Research and Development Project(2018YFC1707100+1 种基金2018YFC1707103)Heilongjiang Touyan Innovation Team Program.
文摘Objective:The fruit stalk of Schisandra chinensis(Turcz.)Baill.(S.chinensis)has been found to contain bioactive components similar to the fruit of S.chinensis.Here,we report a recent discovery about new nortriterpenoids with a novel skeleton and anti-gastric cancer activity,which were isolated from the fruit stalk of S.chinensis.Methods:The chemical components of ethyl acetate extract from 70%ethanol extract from S.chinensis fruit stalk were separated,purified,and identified by liquid chromatography methods(silica gel,ODS,HPLC)and extensive spectroscopic analyses(NMR,IR,UV,MS,CD).Results:Two new nortriterpenoids,schilancitrilactone M and 25-hydroxyl schindilactone D(1 and 2),along with ten known nortriterpenoids(3-12)were isolated from the fruit stalk of S.chinensis.The isolated compounds were tested for their cytotoxic activities against MGC-803 cells,and the results showed that compounds 6-8 possessed significant activities with IC50 of 9.01,11.77,and 2.74μmol/L,respectively.Conclusion:Twelve nortriterpenoids including two new compounds were isolated from the fruit stalk of S.chinensis for the first time.Among them,compounds 6-8 showed significant anti-gastric cancer activities.We postulated that the fruit stalk of S.chinensis could be used as an anti-gastric cancer drug.
基金Fourth Chinese Materia Medica Resources Survey(Z135080000022)。
文摘Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF has anti-inflammatory,protective liver,antioxidant,antibacterial and other pharmacological effects.Ancient prescriptions are commonly used in the treatment of chronic diarrhea and other intestinal diseases and diabetes.Modern clinical pharmacology features of SCF polysaccharide(SCFP)in diabetes,liver diseases,enteritis and other aspects have achieved excellent results.Gut is an important digestive organ of human body,but intestinal diseases are varied,including Crohn′s disease,ulcerative colitis,intestinal flora imbalance,etc..It is a chronic and non-specific inflammatory disease.The disease is persisted for a long time and the incidence rate is expected to rise.Most of the symptoms are recurrent diarrhea,bloody stool and abdominal pain.It is considered by the World Health Organization as a refractory disease.At present,there is little possibility of complete cure,which is closely related to complex environmental factors,eating habits and heredity.In recent years,clinical studies have found that SCFP has a variety of pharmacological effects on intestinal protection.①Reduce inflammatory factors:intestinal mucositis is a common adverse reaction in patients with chemotherapy.The development of mucositis is related to pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL^(-1)β,Interferon-γ(IFN-γ).SCFP can significantly reduce IL-6 TNF-α,IL^(-1)β,and IL-8,as well as the accumulation of T cells in the process of resisting apoptosis,reduce the inflammatory reaction and protect the damage to villi and crypts,improve the symptoms of small intestinal mucositis caused by weight loss and diarrhea.②Promote immunoglobulin A secretion:intestinal mucosal immunity is the first line of defense of the body′s immune system.Its main antibody is secretory immunoglobulin A,which can destroy and phagocytize microorganisms,bacteria and viruses.SCFP can improve intestinal immunity by increasing the number and activity of T lymphocytes,promoting the secretion of secretory immunoglobulin A,and affecting the activity of a variety of cytokines.③Regulation of intestinal flora:the flora in the intestine has the functions of auxiliary nutrient absorption,biological antagonism and immune regulation,and can form a natural barrier for the host's intestine.When the human intestinal flora is disordered,probiotics will be greatly reduced,harmful bacteria will proliferate and destroy the intestinal environment.Under these conditions,the intake of SCFP significantly increased the number of beneficial bacteria such as bifidobacteria and lactobacillus,and significantly decreased the number of conditional pathogens such as enterococcus and escherichia coli,indicating that SCFP can indeed regulate the intestinal disorder caused by lincomycin hydrochloride to a certain extent. This may be because beneficial bacteria in the intestine metabolize polysaccharides produce short chain fatty acids such as lactic acid and acetic acid, which reduces the pH value in the intestine and inhibits the growth of enterococcus and Escherichia coli. In conclusion, SCFP can treat and protect intestinal diseases to a certain extent, which provides a favorable basis for the treatment of intestinal diseases.
文摘Schisandra chinensis (Turcz.) Baill. is a deciduous woody vine, and belongs to the Schisandraceae family. This species is a horticultural plant with edible fruit. It is used medicinally and for making rope. It is distributed in Hebei, Heilongjiang, Jilin, Liaoning, Nei Mongol,
基金Support Program for New Century Excellent Talents in the National Ministry of Education,No. NCET-04-0657Henan Project for cultivation of Innovation Talents in Colleges and Universities No.2004-23
文摘BACKGROUND:The most prominent characteristic of brain aging is decreased learning and memory ability.The functions of learning and memory are closely related to intracerebral acetylcholinesterase(AChE) and monoamine neurotransmitter activity.Previous studies have shown that Schisandra chinensis polysaccharide has an anti-aging effect. OBJECTIVE:To explore the effects of Schisandra chinensis polysaccharide on AChE activity and monoamine neurotransmitter content,as well as learning and memory ability in a D-galactose-induced aging mouse brain model compared with the positive control drug Kangnaoling. DESIGN,TIME AND SETTING:Completely randomized,controlled experiment based on neurobiochemistry was performed at the Pharmacological Laboratory,Henan University of Traditional Chinese Medicine from September to December 2003. MATERIALS:Schisandra chinensis was purchased from Henan Provincial Medicinal Company. Schisandra chinensis polysaccharide was obtained by water extraction and alcohol precipitation. Kangnaoling pellets were provided by Liaoning Tianlong Pharmaceutical(batch No.20030804;state drug permit No.H21023095).A total of 50 six-week-old Kunming mice were randomly divided into five groups:blank control,model,Kangnaoling,high and low dosage Schisandra chinensis polysaccharide groups,with 10 mice per group. METHODS:Mice in the blank control group were subcutaneously injected with 0.5 mL/20 g normal saline into the nape of the neck each day,while the remaining mice were subcutaneously injected with 5%D-galactose saline solution(0.5 mL/20 g) in the nape for 40 days to induce a brain aging model.On day 11,mice in the high and low dosage Schisandra chinensis polysaccharide groups were intragastrically infused with 20 mg/mL and 10 mg/mL Schisandra chinensis polysaccharide solution(0.2 mL/10 g),respectively.Mice from the Kangnaoling group were intragastrically infused with 35 mg/mL Kangnaoling suspension(0.2 ml/10 g),and the mice in the model group were intragastrically infused with the same volume of normal saline(0.2 mL/10 g) once per day for 30 consecutive days. MAIN OUTCOME MEASURES:Two hours after the final administration,pathohistological changes in the cerebral cortex and hippocampus were observed using hematoxylin & eosin staining.AChE activity was detected using chromatometry.Monoamine neurotransmitter content was measured using fluorimetry.Learning and memory was measured using the step down test and darkness avoidance test. RESULTS:Both Schisandra chinensis polysaccharide and Kangnaoling improved pathological injury to the cerebral cortex and hippocampus in a mouse model of brain aging.Compared with the blank control group,AChE activity and content of norepinephrine(NA),dopamine(DA),and 5-hydroxytryptamine(5-HT) were significantly decreased in the model group(P<0.01).In contrast, AChE activity and NA,DA,and 5-HT levels significantly increased in the Kangnaoling and high dosage Schisandra chinensis polysaccharide groups(P<0.01),while NA levels significantly increased in the low dosage Schisandra chinensis polysaccharide group(P<0.01).Drug treatment improved learning and memory abilities(P<0.01 or P<0.05). CONCLUSION:Schisandra chinensis polysaccharide significantly increased levels of central neurotransmitters and improved learning and memory in a mouse model of brain aging.The effects of Schisandra chinensis polysaccharide were equal to that of Kangnaoling pellets.
文摘The essential oil of Schisandra Chinensis(Turc z.) Baill was extracted and analyzed. The essential oil was obtained in a 1.8%(mL/100 g material) yield. Forty eight compounds were isolated and characterized from the oil. The results show that the content(20.42%) of β -pinene of naturally essential oil is the highest among the components of the oil,which has laid a foundation for the futher development and application of the stems of Schisandra Chinensis(Turc z.) Baill .
基金Supported by National Basic Research Program of China(2004CB11750-5)
文摘The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy(HPLC).The sample was extracted exceedingly in the critical limit of CO 2 .The retention time of Schisandrin A and Schisandrin B was reduced,with methano/water(75:25)as mobile phase.The wavelength for detection was 254 nm. The R2 of standard curve was 0.9998 and the relative standard deviation was 2.31%and 3.17%with the recovery of 96.45%and 97.37%,respectively.The result shows that the rate of veracity of this method is higher and it proves that the determination method of Schisandrin A and Schisandrin B in Schisandra chinensis is a feasible method.
基金supported by National Science and Technology Major Project(2016ZX09J161042012ZX09301003-002-001)
文摘OBJECTIVE To study the pharmacokinetics change of schisandra chinensis under the pathological condition of liver dysfunction for safe and rational use of herbal medicines.METHODS The metabolism of four effective lignans from schisandra chinensis(SC),schisandrin,schisantherin A,deoxyshisandrin and γ-schisandrin was studied using microsomes from patients with advanced hepato.cellular carcinoma.In situ intestinal and hepatic perfusions were conducted to clarify the contributions from impairments of gut and liver on the pharmacokinetics of the four schisandra lignans in CCl4-intoxi.cated rats.The metabolism in rat and human liver microsomes and transport in Caco-2 monolayer cell model were studied to reveal the key factors for the in vivo disposition of the four lignans.RESULTS When SC alcoholic extract was orally administrated to CCl4-intoxicated rat for a short term(4 d),the pharmacokinetics of four active SC lignans was significantly changed while its hepatoprotective effect was not obviously observed.The plasma concentrations of the four schisandra lignans were dramatical.ly elevated compared with the control.The Cmax,AUC and MRT were all increased or prolonged signif.icantly while parameter CLz/F was obviously reduced in rat pretreated with CCl4.In hepatic perfusion study and liver microsomes incubation,it was found that the hepatic metabolism of the four lignans was markedly decreased mainly due to the activity reduction of multiple CYP450 isoenzymes involved the metabolism,which,eventually,might lead to the alternation of their pharmacokinetic profiles in CCl4-intoxicated rats or patients with advanced hepatocellular carcinoma.CONCLUSION The pharmacoki.netic studies of SC components in pathological situation of liver dysfunction are expected to provide useful data for rational and safe application of SC preparations in clinic or further pharmacological and toxicological research.
基金Chinese Pharmacopoeia Commission Drug Standardization Research Project(2019Z061).
文摘[Objectives]To study the quality standard of Chinese drug in compound Qianglinaogingsu Tablets(Fructus Schisandrae Chinensis and Coru Cervi Pantotrichum).[Methods]The thin layer chromatography(TLC)differentiation method was established for Fructus Schisandrae Chinensis and Cornu Cervi Pantotrichum.[Results]TLC spots developed were fairly clear,and the blank test showed no interference.[Conclusions]The method is fast,simple and can be used for quality control of Qianglinaoqingsu Tablets.
基金supported by the National Natural Science Foundation of China (32030011,31630071)National Key Research and Development Program of China (2022YFF1301600)Priority Academic Program Development of Jiangsu Higher Education Institutions。
文摘As highly social animals,Indo-Pacific humpback dolphins(Sousa chinensis)exhibit community differentiation.Nevertheless,our understanding of the external and internal factors influencing these dynamics,as well as their spatiotemporal variations,is still limited.In the present study,variations in the social structure of an endangered Indo-Pacific humpback dolphin population in Xiamen Bay,China,were monitored over two distinct periods(2007–2010 and 2017–2019)to analyze the effects of habitat utilization and the composition of individuals within the population.In both periods,the population demonstrated a strikingly similar pattern of social differentiation,characterized by the division of individuals into two main clusters and one small cluster.Spatially,the two primary clusters occupied the eastern and western waters,respectively,although the core distribution area of the eastern cluster shifted further eastward between the two periods.Despite this distribution shift,the temporal stability of the social structure and inter-associations within the eastern cluster remained unaffected.A subset of 16individuals observed in both periods,comprising 51.6%and 43.2%of the population in each respective period,emerged as a foundational element of the social structure and may be responsible for sustaining social structure stability,especially during the 2007–2010 period.These observations suggest that the composition of dominant individuals,an internal factor,had a more substantial influence on the formation of the social network than changes in habitat use,an external factor.Consequently,the study proposes distinct conservation measures tailored to each of the two main clusters.
基金funded by the National Natural Science Foundation of China(Grant Nos.32170236 and 31501333)the Natural Science Foundation of Hebei Province(Grant No.C2020209064)the Youth Foundation of Educational Committee of Hebei Province(Grant No.QN2020139)。
文摘Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by using syntenic analysis combined with phylogenomic analysis of C.chinensis and four other representative genomes from basal and core eudicots,we confirmed that the WGD event in C.chinensis was shared by Aquilegia coerulea and Papaver somniferum L.and quickly occurred after Ranunculales diverged from other eudicots,likely a Ranunculales common tetraploidization(RCT).The synonymous nucleotide substitutions at synonymous sites distribution of syntenic blocks across these genomes showed that the evolutionary rate of the P.somniferum genome is faster than that of the C.chinensis genome by approximately 13.7%,possibly due to Papaveraceaes having an additional special tetraploidization event(PST).After Ks correction,the RCT dated to 115—130 million years ago(MYA),which was close to the divergence of Ranunculaceaes and Papaveraceaes approximately115.45—130.51 MYA.Moreover,we identified homologous genes related to polyploidization and speciation and constructed multiple sequence alignments with different reference genomes.Notably,the event-related subgenomes in the basal genomes all showed genomic fractionation bias,suggesting a likely allopolyploid nature of the RCT,PST and T-Alpha and T-Beta events in Tetracentron sinense.In addition,we detected that the sixteen P450 subfamilies were markedly expanded in the genomes of Ranunculales,and most of them were related to the RCT and PST events.We constructed a new platform for Early Eudicot Comparative Genomic Research(http://www.cgrpoee.top/index.html)to store more information.In summary,our findings support the WGD of C.chinensis shared by Ranunculales,which is likely an allotetraploidization event.This present effort offered new insights into the evolution of key polyploidization events and the genes related to secondary metabolites during the diversification of early eudicots.
基金supported by the National Key R&D Program of China (2021YFC2300900,2021YFC2301300)Academic Promotion Programme of Shandong First Medical University (2019QL006)+2 种基金Natural Science Foundation of Shandong Province (ZR2020QH274)Yunnan Key Research and Development Program (202103AQ100001,202102AA310055)Key Program of Chinese Academy of Sciences (KJZD-SW-L11)。
文摘The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.These similarities have established the tree shrew as a promising experimental model for biomedical research on cancer,infectious diseases,metabolic disorders,and mental health conditions.Herein,we used metatranscriptomic sequencing to analyze plasma,as well as oral and anal swab samples,from 105 healthy asymptomatic tree shrews to identify the presence of potential zoonotic viruses.In total,eight mammalian viruses with complete genomes were identified,belonging to six viral families,including Flaviviridae,Hepeviridae,Parvovirinae,Picornaviridae,Sedoreoviridae,and Spinareoviridae.Notably,the presence of rotavirus was recorded in tree shrews for the first time.Three viruses-hepacivirus 1,parvovirus,and picornavirus-exhibited low genetic similarity(<70%)with previously reported viruses at the whole-genome scale,indicating novelty.Conversely,three other viruses-hepacivirus 2,hepatovirus A and hepevirus-exhibited high similarity(>94%)to known viral strains.Phylogenetic analyses also revealed that the rotavirus and mammalian orthoreovirus identified in this study may be novel reassortants.These findings provide insights into the diverse viral spectrum present in captive Chinese tree shrews,highlighting the necessity for further research into their potential for crossspecies transmission.
文摘The purpose of this project is used for exploring the mechanism of Callistephus chinensis in the treatment of diabetes by network pharmacology and molecular docking methods.The target of Callistephus chinensis was obtained from SwissTargetPrediction database,while the target related to diabetes was obtained from GeneCards and OMIM databases.The target was added in String database to build the protein interaction network.GO biological process enrichment analysis and KEGG pathway enrichment analysis were carried out by Metascape software,then the target-pathway network was constructed.Molecular docking was carried out in Discovery Studio 2016 Client software to verify the binding force of Callistephus chinensis flavonoid compounds with key targets.In this study,10 potential active components were selected from the flavonoid monomer compounds of Callistephus chinensis.1847 biological processes(BP),126 cell compositions(CC)and 256 molecular functions(MF)were obtained by GO enrichment analysis;a total of 194 pathways were involved in KEGG enrichment analysis of 192 cross targets.Network analysis showed that quercetin was the main active component of flavonoids in the treatment of diabetes,AKT1,TNF,VEGFA,EGFR,SRC and other related signals were in relation to the treatment of diabetes.This study showed that Callistephus chinensis flavonoid compounds play a role in the treatment of diabetes by regulating multi-target and multi-pathway.
基金supported by the Achievement Conversion Project of the Ministry of Science and Technology (No.2014GB2B100007)the Science and Technology Project of Education Department of Jilin Province (No. JJKH20180640KJ)。
文摘Schisandra chinensis Turcz.(Baill.) is a plant species with fruits that have been well known in Far Eastern medicine for a long time. It has traditionally been used as a stimulating and fortifying agent in cases of physical exhaustion and to inhibit fatigue.The major bioactive compounds found in S. chinensis are lignans with a dibenzocyclooctadiene skeleton, but little is known about their biosynthesis in plants. S. chinensis is the ideal medicinal plant for studying the biosynthesis of lignans, especially the dibenzocyclooctadiene skeleton. Genomic information for this important herbal plant is unavailable. To better understand the lignan biosynthesis pathway, we generated transcriptome sequences from the fruit during ripening and performed de novo sequence assembly, yielding136 843 unique transcripts with N50 of 1778 bp. Putative functions could be assigned to 41 824 transcripts(51.57%) based on BLAST searches against annotation databases including GO(Gene ontology) and KEGG(Kyoto Encyclopedia of Genes and Genomes). Furthermore, 22 candidate cytochrome P450 genes and 15 candidate dirigent proteins genes that were most likely involved in the lignan biosynthesis pathway were discovered based on transcriptome sequencing of S. chinensis. The genomic data obtained from S. chinensis, especially the identification of putative genes involved in the lignan biosynthesis pathway, will facilitate our understanding of lignan biosynthesis at the molecular level. The lignan metabolite profiles were analyzed by metabolomes, the accumulation patterns of 30 metabolites involved in the lignan pathway were studied. Co-expression network of lignan contents and transcriptional changes showed355 strong correlations(correlation coefficient, R^2 > 0.9) between 21 compounds and 153 transcripts. Furthermore, the comprehensive analysis and characterization of the genes involved in lignan pathways and the metabolite profiles of lignans are expected to provide better insight regarding the diversity of the chemical composition, synthetic characteristics, and regulatory mechanisms of this medical herb.
基金E&T modern center for Natural Products of Liaoning Province of China (2008402021)
文摘Objective To optimize the extract technology of active lignins from the fruits of Schisandra chinensis. Methods The content of schizandrin, gomisin A, and deoxyschizandrin were selected as standards to evaluate the efficiency of smashing tissue extraction (STE). Solid-liquid ratio, extracting times, ethanol concentration, and extracting time were investigated through orthogonal test. Results The optimized conditions for STE were ten times amount of 80% EtOH, extracting for three times, and 2 min for each time. Conclusion STE could obtain relatively higher yield, simplicity of operation, and benefit for environment protection. It could be better choice for the extraction of S. chinensis.
基金the National Natural Science Foundation of China(Nos.81373290,81102483)the Natural Science Foundation of Yunnan Province(No.2012FB178)+1 种基金the Chinese Academy of Sciences grant(No.KSCX2-EW-Q-10)sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry to W.-L.Xiao.
文摘Six new lignans(1-6),as well as five known ones(7-11)were isolated from the leaves and stems of Schisandra chinensis.The structures of 1-6 were established on the basis of spectroscopic methods including 1D-and 2D-NMR techniques and CD experiments.Compound 1 was the first example of naturally occurring N-containing lignans featuring a nicotinoyl group.All the new compounds were evaluated for their anti-HIV-1 activities and showed EC50 values in the range 17.89-138.23μg/mL.
基金Supported by the Major National Science and Technology Project (No. 2018ZX09101-002)。
文摘Objective: To investigate the protective effects of Schisandra chinensis oil(SCEO) against aristolochic acid Ⅰ(AAⅠ)-induced nephrotoxicity in vivo and in vitro and elucidate the underlying mechanism.Methods: C57BL/6 mice were randomly divided into 5 groups according to a random number table, including control group, AAⅠ group, and AAⅠ+SCEO(0.25, 0.5 and 1 g/kg) groups(n=5 per group). Pretreatment with SCEO was done for 2 days by oral administration, while the control and AAⅠ groups were treated with sodium carboxymethyl cellulose. Mice of all groups except for the control group were injected intraperitoneally with AAⅠ(5 mg/kg) from day 3 until day 7. Histopathological examination and apoptosis of kidney tissue were observed by hematoxylin and eosin and TdT-mediated dUTP nick-end labeling(TUNEL) staining, respectively. The levels of serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), blood urea nitrogen(BUN), and serum creatinine(SCr), as well as renal malondialdehyde(MDA), glutathione, r-glutamyl cysteingl+glycine(GSH), and superoxide dismutase(SOD) were analyzed using enzyme-linked immunosorbent assay(ELISA).Expressions of hepatic cytochrome P450 1A1(CYP1A1), CYP1A2, and nad(p)hquinonedehydrogenase1(NQO1) were analyzed using ELISA, quantitative real-time polymerase chain reaction(qPCR) and Western blot,respectively. In vitro, SCEO(40 μg/mL) was added 12 h before treatment with AAⅠ(40 μmol/mL for 48 h) in human renal proximal tubule cell line(HK-2), then apoptosis and reactive oxygen species(ROS) were analyzed by flow cytometry. Results: SCEO 0.5 and 1 g/kg ameliorated histopathological changes and TUNEL+ staining in the kidney tissues of mice with AAⅠ-induced nephrotoxicity, and reduced serum levels of ALT, AST, BUN and SCr(P<0.01 or P<0.05). SCEO 0.5 and 1 g/kg alleviated the ROS generation in kidney, containing MDA,GSH and SOD(P<0.01 or P<0.05). SCEO 1 g/kg increased the expressions of CYP1A1 and CYP1A2 and decreased NQO1 level in the liver tissues(P<0.01 or P<0.05). Besides, in vitro studies also demonstrated that SCEO 40 μg/mL inhibited apoptosis and ROS generation(P<0.05 or P<0.01). Conclusions: SCEO can alleviate AAⅠ-induced kidney damage both in vivo and in vitro. The protective mechanism may be closely related to the regulation of metabolic enzymes, thereby inhibiting apoptosis and ROS production.
基金supported by the National Key R&D Program of China(No.2017YFC1701200).
文摘Phenylalanine ammonia-lyase(PAL),which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid,is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes.Schisandra chinensis,a woody vine plant belonging to the family of Magnoliaceae,is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity.However,the functional role of PAL in the biosynthesis of lignan is relatively limited,compared with those in lignin and flavonoids biosynthesis.Therefore,it is essential to clone and characterize the PAL genes from this valuable medicinal plant.In this study,molecular cloning and characterization of three PAL genes(ScPAL1−3)from S.chinensis was carried out.ScPALs were cloned using RACE PCR.The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis.In order to determine their catalytic activity,recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli(BL21-DE3),followed by Ni-affinity purification.The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds.The optimal temperature,pH value and effects of different metal ions were determined.Vmax,Kcat and Km values were determined under the optimal conditions.The expression of three ScPALs in different tissues was also determined.Our work provided essential information for the function of ScPALs.
基金supported by the grants of Scientific Research Foundation for the Returned Overseas Chinese Scholars(Jilin Province,2015)Jilin Science&Technology Development Plan(No.20160209008YY)the Program for the Young Top-notch and Innovative Talents of Jilin Agricultural University(2016-2018)
文摘The present study was designed to evaluate protective activity of an ethanol extract of the stems of Schisandra chinensis(SCE) and explore its possible molecular mechanisms on acetaminophen(APAP) induced hepatotoxicity in a mouse model. The results of HPLC analysis showed that the main components of SCE included schisandrol A, schisandrol B, deoxyschisandrin, schisan--drin B, and schisandrin C and their contents were 5.83, 7.11, 2.13, 4.86, 0.42 mg·g^(-1), respectively. SCE extract was given for 7 con--secutive days before a single hepatotoxic dose of APAP(250 mg·kg^(-1)) was injected to mice. Our results showed that SCE pretreatment ameliorated liver dysfunction and oxidative stress, which was evidenced by significant decreases in aspartate transaminase(AST), alanine aminotransferase(ALT), malondialdehyde(MDA) contents and elevations in reduced glutathione(GSH) and superoxide dismutase(SOD) levels. These findings were associated with the result that the SCE pretreatment significantly decreased expression levels of 4-hydroxynonenal(4-HNE) and 3-nitrotyrosine(3-NT). SCE also significantly decreased the expression levels of Bax, mitogen-activated protein kinase(MAPK), and cleaved caspase-3 by APAP exposure. Furthermore, supplementation with SCE suppressed the expression levels of inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2), suggesting alleviation of inflammatory response. In summary, these findings from the present study clearly demonstrated that SCE exerted significant alleviation in APAP-induced oxidative stress, inflammation and apoptosis mainly via regulating MAPK and caspase-3 signaling pathways.
