Ischemic accumulation of succinate causes cerebral damage by excess production of reactive oxygen species. However, it is unknown whether ischemic accumulation of succinate affects neural stem cell proliferation. In t...Ischemic accumulation of succinate causes cerebral damage by excess production of reactive oxygen species. However, it is unknown whether ischemic accumulation of succinate affects neural stem cell proliferation. In this study, we established a rat model of cerebral ischemia/reperfusion injury by occlusion of the middle cerebral artery. We found that succinate levels increased in serum and brain tissue(cortex and hippocampus) after ischemia/reperfusion injury. Oxygen-glucose deprivation and reoxygenation stimulated primary neural stem cells to produce abundant succinate. Succinate can be converted into diethyl succinate in cells. Exogenous diethyl succinate inhibited the proliferation of mouse-derived C17.2 neural stem cells and increased the infarct volume in the rat model of cerebral ischemia/reperfusion injury. Exogenous diethyl succinate also increased the succinylation of the Rho family GTPase Cdc42 but repressed Cdc42 GTPase activity in C17.2 cells. Increasing Cdc42 succinylation by knockdown of the desuccinylase Sirt5 also inhibited Cdc42 GTPase activity in C17.2 cells. Our findings suggest that ischemic accumulation of succinate decreases Cdc42 GTPase activity by induction of Cdc42 succinylation, which inhibits the proliferation of neural stem cells and aggravates cerebral ischemia/reperfusion injury.展开更多
To explore the effect of Buyang Huanwu Decoction (BHD) on platelet activating factor (PAF) content in arterial blood pre- and post-arterial thrombosis in rats, male Wistar rats were randomly divided into 3 groups, the...To explore the effect of Buyang Huanwu Decoction (BHD) on platelet activating factor (PAF) content in arterial blood pre- and post-arterial thrombosis in rats, male Wistar rats were randomly divided into 3 groups, the medicine group treated with BHD, the control group with dexamethasone liquid, and the blank group with distilled water. Oral administration was given for 14 consecutive days, once daily. Model of arterial thrombosis was established in the animals 2 hours after final medication, the blood content of PAF, dry weight (DW) and occlusion time (OT) of thrombus, and dry weight of thrombus/body weight (TW/BW) ratio were observed. Results indicated that BHD could markedly lower the arterial blood content of PAF after thrombosis, increase the OT of thrombus, reduce the dry weight of thrombus and the TW/BW ratio (P<0.05). It is suggested that BHD can inhibit the pathologic PAF metabolism and formation and development of arterial thrombus.展开更多
Galangal(Alpinia officinaruim Hance)is the rhizome of the perennial herb belonging to Zingiberaceae family.There are many active components in galangal,such as volatile oil,flavonoids,terpenoids,phenylpropanoids and g...Galangal(Alpinia officinaruim Hance)is the rhizome of the perennial herb belonging to Zingiberaceae family.There are many active components in galangal,such as volatile oil,flavonoids,terpenoids,phenylpropanoids and glycosides,among which the content of volatile oil is higher.The bioactivities of galangal volatile oil on health effect includesanti-inflammatory,anti-hypertension,anti-oxidation and prevention of cardiovascular diseases.Cardiovascular disease(CVD)is a kind of diseases related to circulatory system,which is also called circulatory system diseases.Over the past decade,the number of people dying from CVD has increased by 12.5%worldwide,and it is now the leading cause of human death worldwide.Studies have shown that galangal volatile oil has good pharmacological effects in treating CVD.①Regulation of glucose and lipid metabolism:studies have found that abnormal lipid metabolism can lead to obesity,diabetes,CVD and other diseases.The serum total triglyceride(TG)content in liver and serum will increase in patients with abnormal fat metabolism.The results showed that the volatile oil of galangal could increase the excretion of neutral cholesterol,significantly reduce liver TG and serum TG,and thus regulate glucose and lipid metabolism,prevent lipid deposition and prevent CVD.②Improving insulin resistance(IR):inhibition of inflammatory cytokines such as IL^(-1),IL-6 activation and expression of TNF-α,improves IR,thereby protecting myocardium from IR-mediated damage.Through the establishment of endothelial cell injury model induced by high glucose in vitro,it was found that the volatile oil of galangal can significantly reduce the secretion of pro-inflammatory cytokines TNF-αand IL-8,and inhibit the expression of ICAM-1 and VCAM-1 induced by high glucose,suggesting that it has protective effect on endothelial dysfunction and inflammation induced by high glucose.③Regulate blood oxygenation:during acute myocardial hypoxia,the activity of free radical scavenging system is decreased,and oxygen free radicals are produced in large quantity,which reacts with unsaturated fatty acids on the cell membrane and forms lipid peroxidation,resulting in myocardial structural damage.The results showed that the water extract of Galangal could reduce the content of MDA in blood and protect the SOD activity of ischemic and hypoxic myocardium.④Protective effect of vascular endothelial cells(ES):ES injury is the pathological basis of some cardiovascular diseases.The results showed that the volatile oil of galangal had a protective effect on ES apoptosis.Compared with the morphology and activity of ES treated with oxidized LDL,galangal volatile oil could ameliorate these morphological changes and improve cell viability.⑤Antiplatelet agglutination:inhibit platelet aggregation and thromboxane release,improve blood circulation,and have obvious anti-thrombotic effect,which has a good effect on the treatment and prevention of cardiovascular diseases.The results showed that the volatile oil of galangal had inhibitory effect on platelet aggregation and anticoagulant effect.In conclusion,the volatile oil of galangal can be used to prevent and treat cardiovascular diseases.Based on the mechanism of CVD,this study summarized the role of the essential oil of Alpinia officinaruim in CVD,providing basis for the clinical application of alpiniaofficinalis essential oil in the prevention and treatment of CVD and the development of new drugs.展开更多
We previously found that monocyte locomotion inhibitory factor has a neuroprotective effect on ischemic brain injury during the acute phase of stro ke.Therefore,we modified the structure of an anti-inflammato ry monoc...We previously found that monocyte locomotion inhibitory factor has a neuroprotective effect on ischemic brain injury during the acute phase of stro ke.Therefore,we modified the structure of an anti-inflammato ry monocyte locomotion inhibitory factor peptide to construct an active cyclic peptide—Cyclo(MQCNS)(LZ-3)—and investigated its effects on ischemic stroke.In this study,we established a rat model of ischemic stroke by occluding the middle cerebral artery and then administered LZ-3(2 or 4 mg/kg) via the tail vein for 7 consecutive days.Our res ults showed that LZ-3(2 or 4 mg/kg) substantially decreased infarct volu m e,reduced co rtical ne rve cell death,improved neurological function,reduced cortical and hippocampal injury,and decreased the levels of inflammatory factors in the blood and brain tissues.In a well-diffe rentiated,oxygen-glucose deprivation/reoxygenation-induced BV2 cell model of poststroke,LZ-3(100 μM) inhibited the JAK1-STAT6 signaling pathway.LZ-3 regulated microglia/macrophage polarization from the M1 to the M2 type and inhibited microglia/macrophage phagocytosis and migration via the JAK1/STAT6 signaling pathway.To conclude,LZ-3 regulates microglial activation by inhibiting the JAK1/STAT6 sign aling pathway and improves functional recovery post-stroke.展开更多
1.Entropy and its implications in human disease and aging Entropy,which is a thermodynamic property or an interpreta-tion of the second law of thermodynamics,was first defined in 1865 by the German physicist Rudolph C...1.Entropy and its implications in human disease and aging Entropy,which is a thermodynamic property or an interpreta-tion of the second law of thermodynamics,was first defined in 1865 by the German physicist Rudolph Clausius[1].It eventually evolved into a general scientific concept that is of universal and paramount importance in three aspects.First,entropy is a measur-able physical property that is commonly associated with a state of chaos,disorderliness,randomness,or uncertainty of any systems[2].Second,entropy is a measure of the amount of energy that is unavailable to do work[2].Third,the universe or an isolated sys-tem always obeys“the principle of entropy increase”that irre-versible or spontaneous processes can occur only in the direction of entropy increase—that is,the direction of increasing chaos,dis-order,randomness,or uncertainty[2].展开更多
Objective:To explore the molecular mechanism by which cordycepin inhibits cell proliferation and induces apoptosis of human colorectal cancer cells.Methods:Cell counting and MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carbo...Objective:To explore the molecular mechanism by which cordycepin inhibits cell proliferation and induces apoptosis of human colorectal cancer cells.Methods:Cell counting and MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium,inner salt) method were used to monitor the effects of cordycepin on cell proliferation.Flow cytometry(FCM) was used to analyze the effects of cordycepin on the cell cycle progress.Annexin V-fluorescein isothiocyanate(FITC) analysis was used to detect apoptosis at a very early stage.Caspase-Glo was used to determine caspase activity and Western blot was used to measure protein expression levels of c-Jun N-terminal kinase(JNK),p38,and Bcl-2 pro-apoptosis family.Results:The numbers of viable SW480 and SW620 cells and the proliferation of these cells were significantly reduced with increases in cordycepin concentration(P<0.01).The cell cycle progression of SW480 and SW620 was arrested at the G0/G1 phase by the addition of cordycepin,and apoptosis rates of cordycepin treatments were increased compared with the control group.Cordycepin-treated cells showed phosphatidylserine valgus,suggesting the existence of early apoptosis.Caspase-3/7 and-9 activity significantly increased and the protein expression levels of JNK,p38,and Bax,Bid,Bim,and Puma from Bcl-2 pro-apoptosis molecules also increased after the treatment with cordycepin.Conclusions:Cordycepin can inhibit SW480 and SW620 cell proliferation and induce apoptosis.Apoptosis might be induced by enhancing JNK and p38 kinase activity and increasing the protein expression of Bcl-2 pro-apoptotic molecules.展开更多
Objective To study the effect of autotoxicity and the co-effect of autotoxicity and soil microbes from continuous cropping soil on Angelica sinensis growth, root yield and content of essential oils, and soil microbal ...Objective To study the effect of autotoxicity and the co-effect of autotoxicity and soil microbes from continuous cropping soil on Angelica sinensis growth, root yield and content of essential oils, and soil microbal population. Methods The pot experiments were conducted upon A. sinensis seedlings in continuous cropping soil. At the different growth stages, we determined the seedlings in growth parameters, root yield, content of essential oils, ethanol extract, and cultivable microbial populations in rhizosphere soil. Results A. sinensis seedlings were significantly inhibited in growth, root yield and quality. Compared with the control, the composition and structure of soil microbes were changed and the diversity indexes of bacteria functional groups were reduced in rhizosphere soil of A. sinensis. A. sinensis cropping problems were more seriously after the treatment with combination of autotoxicity and soil microbes than with autotoxicity alone. Conclusion The autotoxicity and soil microbes from continuous cropping soil of A. sinensis could cause the continuous cropping obstacle together.展开更多
OBJECTIVE: To quantitatively evaluate the therapeutic effects of Red Liriope platyphylla(RLP) on atopic dermatitis(AD), alterations in the luciferase(Luc) signal and general phenotype biomarkers were compared in phtha...OBJECTIVE: To quantitatively evaluate the therapeutic effects of Red Liriope platyphylla(RLP) on atopic dermatitis(AD), alterations in the luciferase(Luc) signal and general phenotype biomarkers were compared in phthalic anhydride(PA) treated Interleukin-4(IL-4)/Luc/Consensus non-coding sequence-1(CNS-1) transgenic(Tg) mice following treatment with aqueous extract of RLP(AEt RLP) for4 weeks.METHODS: Alterations in AD phenotypes were measured in IL-4/Luc/CNS-1 Tg mice following treatment with AEt RLP using inflammation parameter analysis, bioluminescence imaging analysis, histological analysis, reverse transcription-polymerase chain reaction, enzyme linked immunosorbent assay and Western blot analysis.RESULTS: RLP contained high concentrations of total phenolic compounds, total flavonoid compounds and 5-HNE related to AD therapy. The Luc signal was only detected in the abdominal region and the submandibular lymph node(SL), mesenteric lymph node(ML), thymus and pancreas of the PA treated group. This signal was significantly decreased by 28%-73% throughout the body and in the four organs in PA + AEt RLP treated group. Furthermore, the lymph node weight, immunoglobulin E concentration and dermal thickness were decreased by 37%-67% in the PA + AEt RLP treated group.CONCLUSION: Our findings suggest that the therapeutic effect of AEt RLP on PA induced AD could be successfully quantified by comparison of Luc signals and AD phenotype markers in IL-4/Luc/CNS-1Tg mice, and that the Luc signal was as sensitive as the general AD phenotypes, enabling detection of effects without euthanasia.展开更多
The purpose of this work was to construct the plasmid that could direct the synthesis of siRNA-like transcripts and thus mediate strong and specific repression of human heat shock protein 90β(Hsp90β)gene expression ...The purpose of this work was to construct the plasmid that could direct the synthesis of siRNA-like transcripts and thus mediate strong and specific repression of human heat shock protein 90β(Hsp90β)gene expression and to compare the transfection efficiency of the plasmids in varying conditions of transfection.Three 64 nt oligos corresponding to different regions of the target gene were chemically synthesized and annealed and were then ligated with pSUPER EGFP1 plasmid and double-digested with HindIII and BglII.Recombinant plasmids were transformed into Escherichia coli,DH5a,and the colonies were picked and grown in the Amp-agarose.The presence of positive clones was checked by the means of endodigestion and sequencing.Three cell strains,HepG2,Human umbilicus vein endothelium cells(HUVEC)and HeK293,were cultured.Then the plasmids were transfected into the cells at different ratios of plasmid to Lipofectamine.The transfection efficiency was measured by detection of enhanced green fluorescence protein(EGFP).The presence of positive recombinant clones were verified by double-digestion and sequencing.The bases inserted into the plasmids were correct and the positive colonies were named pSuper-Hsp90β1,pSuper-Hsp90β2 and pSuper-Hsp90β3.After optimizing the ratio of plasmid to Lipofectamine,we achieved high transfection efficiency in HeK293 cells.Transfection efficiency was still low in the HepG2 cells.In conclusion,the si-RNA-synthesizing plasmids targeting Hsp90βwere constructed and transfected into cells with different transfection efficiency.展开更多
基金supported by the National Natural Science Foundation of China,No. 81671164 (to SHQ)the Natural Science Foundation of Jiangsu Province of China,No. BK20211348 (to SHQ)Xuzhou Basic Research Program,No. KC21030 (to LYH)。
文摘Ischemic accumulation of succinate causes cerebral damage by excess production of reactive oxygen species. However, it is unknown whether ischemic accumulation of succinate affects neural stem cell proliferation. In this study, we established a rat model of cerebral ischemia/reperfusion injury by occlusion of the middle cerebral artery. We found that succinate levels increased in serum and brain tissue(cortex and hippocampus) after ischemia/reperfusion injury. Oxygen-glucose deprivation and reoxygenation stimulated primary neural stem cells to produce abundant succinate. Succinate can be converted into diethyl succinate in cells. Exogenous diethyl succinate inhibited the proliferation of mouse-derived C17.2 neural stem cells and increased the infarct volume in the rat model of cerebral ischemia/reperfusion injury. Exogenous diethyl succinate also increased the succinylation of the Rho family GTPase Cdc42 but repressed Cdc42 GTPase activity in C17.2 cells. Increasing Cdc42 succinylation by knockdown of the desuccinylase Sirt5 also inhibited Cdc42 GTPase activity in C17.2 cells. Our findings suggest that ischemic accumulation of succinate decreases Cdc42 GTPase activity by induction of Cdc42 succinylation, which inhibits the proliferation of neural stem cells and aggravates cerebral ischemia/reperfusion injury.
文摘To explore the effect of Buyang Huanwu Decoction (BHD) on platelet activating factor (PAF) content in arterial blood pre- and post-arterial thrombosis in rats, male Wistar rats were randomly divided into 3 groups, the medicine group treated with BHD, the control group with dexamethasone liquid, and the blank group with distilled water. Oral administration was given for 14 consecutive days, once daily. Model of arterial thrombosis was established in the animals 2 hours after final medication, the blood content of PAF, dry weight (DW) and occlusion time (OT) of thrombus, and dry weight of thrombus/body weight (TW/BW) ratio were observed. Results indicated that BHD could markedly lower the arterial blood content of PAF after thrombosis, increase the OT of thrombus, reduce the dry weight of thrombus and the TW/BW ratio (P<0.05). It is suggested that BHD can inhibit the pathologic PAF metabolism and formation and development of arterial thrombus.
基金supported by National Natural Science Foundation of China(No.302700005)the Ministry of Science and Technology of the People's Republic of China(MOST,No.2007Aa021405).
文摘Galangal(Alpinia officinaruim Hance)is the rhizome of the perennial herb belonging to Zingiberaceae family.There are many active components in galangal,such as volatile oil,flavonoids,terpenoids,phenylpropanoids and glycosides,among which the content of volatile oil is higher.The bioactivities of galangal volatile oil on health effect includesanti-inflammatory,anti-hypertension,anti-oxidation and prevention of cardiovascular diseases.Cardiovascular disease(CVD)is a kind of diseases related to circulatory system,which is also called circulatory system diseases.Over the past decade,the number of people dying from CVD has increased by 12.5%worldwide,and it is now the leading cause of human death worldwide.Studies have shown that galangal volatile oil has good pharmacological effects in treating CVD.①Regulation of glucose and lipid metabolism:studies have found that abnormal lipid metabolism can lead to obesity,diabetes,CVD and other diseases.The serum total triglyceride(TG)content in liver and serum will increase in patients with abnormal fat metabolism.The results showed that the volatile oil of galangal could increase the excretion of neutral cholesterol,significantly reduce liver TG and serum TG,and thus regulate glucose and lipid metabolism,prevent lipid deposition and prevent CVD.②Improving insulin resistance(IR):inhibition of inflammatory cytokines such as IL^(-1),IL-6 activation and expression of TNF-α,improves IR,thereby protecting myocardium from IR-mediated damage.Through the establishment of endothelial cell injury model induced by high glucose in vitro,it was found that the volatile oil of galangal can significantly reduce the secretion of pro-inflammatory cytokines TNF-αand IL-8,and inhibit the expression of ICAM-1 and VCAM-1 induced by high glucose,suggesting that it has protective effect on endothelial dysfunction and inflammation induced by high glucose.③Regulate blood oxygenation:during acute myocardial hypoxia,the activity of free radical scavenging system is decreased,and oxygen free radicals are produced in large quantity,which reacts with unsaturated fatty acids on the cell membrane and forms lipid peroxidation,resulting in myocardial structural damage.The results showed that the water extract of Galangal could reduce the content of MDA in blood and protect the SOD activity of ischemic and hypoxic myocardium.④Protective effect of vascular endothelial cells(ES):ES injury is the pathological basis of some cardiovascular diseases.The results showed that the volatile oil of galangal had a protective effect on ES apoptosis.Compared with the morphology and activity of ES treated with oxidized LDL,galangal volatile oil could ameliorate these morphological changes and improve cell viability.⑤Antiplatelet agglutination:inhibit platelet aggregation and thromboxane release,improve blood circulation,and have obvious anti-thrombotic effect,which has a good effect on the treatment and prevention of cardiovascular diseases.The results showed that the volatile oil of galangal had inhibitory effect on platelet aggregation and anticoagulant effect.In conclusion,the volatile oil of galangal can be used to prevent and treat cardiovascular diseases.Based on the mechanism of CVD,this study summarized the role of the essential oil of Alpinia officinaruim in CVD,providing basis for the clinical application of alpiniaofficinalis essential oil in the prevention and treatment of CVD and the development of new drugs.
文摘We previously found that monocyte locomotion inhibitory factor has a neuroprotective effect on ischemic brain injury during the acute phase of stro ke.Therefore,we modified the structure of an anti-inflammato ry monocyte locomotion inhibitory factor peptide to construct an active cyclic peptide—Cyclo(MQCNS)(LZ-3)—and investigated its effects on ischemic stroke.In this study,we established a rat model of ischemic stroke by occluding the middle cerebral artery and then administered LZ-3(2 or 4 mg/kg) via the tail vein for 7 consecutive days.Our res ults showed that LZ-3(2 or 4 mg/kg) substantially decreased infarct volu m e,reduced co rtical ne rve cell death,improved neurological function,reduced cortical and hippocampal injury,and decreased the levels of inflammatory factors in the blood and brain tissues.In a well-diffe rentiated,oxygen-glucose deprivation/reoxygenation-induced BV2 cell model of poststroke,LZ-3(100 μM) inhibited the JAK1-STAT6 signaling pathway.LZ-3 regulated microglia/macrophage polarization from the M1 to the M2 type and inhibited microglia/macrophage phagocytosis and migration via the JAK1/STAT6 signaling pathway.To conclude,LZ-3 regulates microglial activation by inhibiting the JAK1/STAT6 sign aling pathway and improves functional recovery post-stroke.
文摘1.Entropy and its implications in human disease and aging Entropy,which is a thermodynamic property or an interpreta-tion of the second law of thermodynamics,was first defined in 1865 by the German physicist Rudolph Clausius[1].It eventually evolved into a general scientific concept that is of universal and paramount importance in three aspects.First,entropy is a measur-able physical property that is commonly associated with a state of chaos,disorderliness,randomness,or uncertainty of any systems[2].Second,entropy is a measure of the amount of energy that is unavailable to do work[2].Third,the universe or an isolated sys-tem always obeys“the principle of entropy increase”that irre-versible or spontaneous processes can occur only in the direction of entropy increase—that is,the direction of increasing chaos,dis-order,randomness,or uncertainty[2].
基金supported by the Department of Education of Zhejiang Province(No. Y200804636)the Department of Science and Tech-nology of Zhejiang Province(Nos. 2008C23049,2007C23027,and2009C33081)the Natural Science Foundation of Zhejiang Province(No. Y206174),China
文摘Objective:To explore the molecular mechanism by which cordycepin inhibits cell proliferation and induces apoptosis of human colorectal cancer cells.Methods:Cell counting and MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium,inner salt) method were used to monitor the effects of cordycepin on cell proliferation.Flow cytometry(FCM) was used to analyze the effects of cordycepin on the cell cycle progress.Annexin V-fluorescein isothiocyanate(FITC) analysis was used to detect apoptosis at a very early stage.Caspase-Glo was used to determine caspase activity and Western blot was used to measure protein expression levels of c-Jun N-terminal kinase(JNK),p38,and Bcl-2 pro-apoptosis family.Results:The numbers of viable SW480 and SW620 cells and the proliferation of these cells were significantly reduced with increases in cordycepin concentration(P<0.01).The cell cycle progression of SW480 and SW620 was arrested at the G0/G1 phase by the addition of cordycepin,and apoptosis rates of cordycepin treatments were increased compared with the control group.Cordycepin-treated cells showed phosphatidylserine valgus,suggesting the existence of early apoptosis.Caspase-3/7 and-9 activity significantly increased and the protein expression levels of JNK,p38,and Bax,Bid,Bim,and Puma from Bcl-2 pro-apoptosis molecules also increased after the treatment with cordycepin.Conclusions:Cordycepin can inhibit SW480 and SW620 cell proliferation and induce apoptosis.Apoptosis might be induced by enhancing JNK and p38 kinase activity and increasing the protein expression of Bcl-2 pro-apoptotic molecules.
基金Natural Science Foundation of China(31060182)Agricultural Biotechnology Research and Application Development of Gansu province(GNSW-2010-18)
文摘Objective To study the effect of autotoxicity and the co-effect of autotoxicity and soil microbes from continuous cropping soil on Angelica sinensis growth, root yield and content of essential oils, and soil microbal population. Methods The pot experiments were conducted upon A. sinensis seedlings in continuous cropping soil. At the different growth stages, we determined the seedlings in growth parameters, root yield, content of essential oils, ethanol extract, and cultivable microbial populations in rhizosphere soil. Results A. sinensis seedlings were significantly inhibited in growth, root yield and quality. Compared with the control, the composition and structure of soil microbes were changed and the diversity indexes of bacteria functional groups were reduced in rhizosphere soil of A. sinensis. A. sinensis cropping problems were more seriously after the treatment with combination of autotoxicity and soil microbes than with autotoxicity alone. Conclusion The autotoxicity and soil microbes from continuous cropping soil of A. sinensis could cause the continuous cropping obstacle together.
基金supported by grants to Dr. Dae Youn Hwang from the Korea Institute of Planning Evaluation for Technology of Food, Agriculture, Forestry and Fisheries (116027-032-HD030)
文摘OBJECTIVE: To quantitatively evaluate the therapeutic effects of Red Liriope platyphylla(RLP) on atopic dermatitis(AD), alterations in the luciferase(Luc) signal and general phenotype biomarkers were compared in phthalic anhydride(PA) treated Interleukin-4(IL-4)/Luc/Consensus non-coding sequence-1(CNS-1) transgenic(Tg) mice following treatment with aqueous extract of RLP(AEt RLP) for4 weeks.METHODS: Alterations in AD phenotypes were measured in IL-4/Luc/CNS-1 Tg mice following treatment with AEt RLP using inflammation parameter analysis, bioluminescence imaging analysis, histological analysis, reverse transcription-polymerase chain reaction, enzyme linked immunosorbent assay and Western blot analysis.RESULTS: RLP contained high concentrations of total phenolic compounds, total flavonoid compounds and 5-HNE related to AD therapy. The Luc signal was only detected in the abdominal region and the submandibular lymph node(SL), mesenteric lymph node(ML), thymus and pancreas of the PA treated group. This signal was significantly decreased by 28%-73% throughout the body and in the four organs in PA + AEt RLP treated group. Furthermore, the lymph node weight, immunoglobulin E concentration and dermal thickness were decreased by 37%-67% in the PA + AEt RLP treated group.CONCLUSION: Our findings suggest that the therapeutic effect of AEt RLP on PA induced AD could be successfully quantified by comparison of Luc signals and AD phenotype markers in IL-4/Luc/CNS-1Tg mice, and that the Luc signal was as sensitive as the general AD phenotypes, enabling detection of effects without euthanasia.
基金The study was supported by grants from the National Excellent Doctorate Dissertation Author Specific Foundation Program of China(No.200156)the National Natural Science Foundation of China(Grant No.30470988).
文摘The purpose of this work was to construct the plasmid that could direct the synthesis of siRNA-like transcripts and thus mediate strong and specific repression of human heat shock protein 90β(Hsp90β)gene expression and to compare the transfection efficiency of the plasmids in varying conditions of transfection.Three 64 nt oligos corresponding to different regions of the target gene were chemically synthesized and annealed and were then ligated with pSUPER EGFP1 plasmid and double-digested with HindIII and BglII.Recombinant plasmids were transformed into Escherichia coli,DH5a,and the colonies were picked and grown in the Amp-agarose.The presence of positive clones was checked by the means of endodigestion and sequencing.Three cell strains,HepG2,Human umbilicus vein endothelium cells(HUVEC)and HeK293,were cultured.Then the plasmids were transfected into the cells at different ratios of plasmid to Lipofectamine.The transfection efficiency was measured by detection of enhanced green fluorescence protein(EGFP).The presence of positive recombinant clones were verified by double-digestion and sequencing.The bases inserted into the plasmids were correct and the positive colonies were named pSuper-Hsp90β1,pSuper-Hsp90β2 and pSuper-Hsp90β3.After optimizing the ratio of plasmid to Lipofectamine,we achieved high transfection efficiency in HeK293 cells.Transfection efficiency was still low in the HepG2 cells.In conclusion,the si-RNA-synthesizing plasmids targeting Hsp90βwere constructed and transfected into cells with different transfection efficiency.
