BACKGROUND Cryptotanshinone(CPT)has wide biological functions,including anti-oxidative,antifibrosis,and anti-inflammatory properties.However,the effect of CPT on hepatic fibrosis is unknown.AIM To investigate the effe...BACKGROUND Cryptotanshinone(CPT)has wide biological functions,including anti-oxidative,antifibrosis,and anti-inflammatory properties.However,the effect of CPT on hepatic fibrosis is unknown.AIM To investigate the effects of CPT treatment on hepatic fibrosis and its underlying mechanism of action.METHODS Hepatic stellate cells(HSCs)and normal hepatocytes were treated with different concentrations of CPT and salubrinal.The CCK-8 assay was used to determine cell viability.Flow cytometry was used to measure apoptosis and cell cycle arrest.Reverse transcription polymerase chain reaction(RT-PCR)and Western blot analyses were used to measure mRNA levels and protein expression of endoplasmic reticulum stress(ERS)signaling pathway related molecules,respectively.Carbon tetrachloride(CCL4)was used to induce in vivo hepatic fibrosis in mice.Mice were treated with CPT and salubrinal,and blood and liver samples were collected for histopathological examination.RESULTS We found that CPT treatment significantly reduced fibrogenesis by modulating the synthesis and degradation of the extracellular matrix in vitro.CPT inhibited cell proliferation and induced cell cycle arrest at the G2/M phase in cultured HSCs.Furthermore,we found that CPT promoted apoptosis of activated HSCs by upregulating expression of ERS markers(CHOP and GRP78)and activating ERS pathway molecules(PERK,IRE1α,and ATF4),which were inhibited by salubrinal.Inhibition of ERS by salubrinal partially eliminated the therapeutic effect of CPT in our CCL4-induced hepatic fibrosis mouse model.CONCLUSION CPT can promote apoptosis of HSCs and alleviate hepatic fibrosis through modulating the ERS pathway,which represents a promising strategy for treating hepatic fibrosis.展开更多
Objective:To evaluate the neuroprotective effect of cryptotanshinone against cladribine-induced cognitive impairment in rats.Methods:Rats were administered with cladribine(1 mg/kg,p.o.)and cryptotanshinone(10 and 20 m...Objective:To evaluate the neuroprotective effect of cryptotanshinone against cladribine-induced cognitive impairment in rats.Methods:Rats were administered with cladribine(1 mg/kg,p.o.)and cryptotanshinone(10 and 20 mg/kg,i.p.)for four weeks.Behavioral tests such as Morris water maze and elevated plus maze were conducted to check memory impairment caused by cladribine.On day 29,all rats were sacrificed,and the brains were separated for estimation of neuroinflammatory factors,biochemical parameters,neurotransmitters,Aβ(1-42),blood-brain barrier permeability,nuclear factor erythroid 2-related factor 2(Nrf2),and brain-derived neurotrophic factor(BDNF).Results:Treatment with cryptotanshinone dose-dependently enhanced spatial memory,improved the levels of neurotransmitter and antioxidant enzymes,and suppressed proinflammatory cytokine release.Cryptotanshinone also decreased Aβ(1-42)accumulation and increased the levels of Nrf2 and BDNF in the hippocampus.Additionally,the histopathological results showed that cryptotanshinone reduced cladribine-induced neuronal death in the hippocampus.Conclusions:Cryptotanshinone exhibits a promising neuroprotective effect against cladribine-induced cognitive impairment in preclinical studies,and may be a potential phytochemical for the treatment and management of cognitive impairment.展开更多
BACKGROUND Approximately 90%of new cases of noncardiac gastric cancer(GC)are related to Helicobacter pylori(H.pylori),and cytotoxin-associated gene A(CagA)is one of the main pathogenic factors.Recent studies have show...BACKGROUND Approximately 90%of new cases of noncardiac gastric cancer(GC)are related to Helicobacter pylori(H.pylori),and cytotoxin-associated gene A(CagA)is one of the main pathogenic factors.Recent studies have shown that the pharmacological effects of cryptotanshinone(CTS)can be used to treat a variety of tumors.However,the effects of CTS on H.pylori,especially CagA+strain-induced gastric mucosal lesions,on the development of GC is unknown.AIM To assess the role of CTS in CagA-induced proliferation and metastasis of GC cells,and determine if CagA+H.pylori strains causes pathological changes in the gastric mucosa of mice.METHODS The effects of CTS on the proliferation of GC cells were assessed using the Cell Counting Kit-8(CCK-8)assay,and the abnormal growth,migration and invasion caused by CagA were detected by CCK-8 and transwell assays.After transfection with pSR-HA-CagA and treatment with CTS,proliferation and metastasis were evaluated by CCK-8 and transwell assays,respectively,and the expression of Src homology 2(SH2)domain–containing phosphatase 2(SHP2)and phosphorylated SHP2(p-SHP2)was detected using western blotting in AGS cells.The enzymelinked immunosorbent assay was used to determine the immunoglobulin G(IgG)level against CagA in patient serum.Mice were divided into four groups and administered H.pylori strains(CagA+or CagA-)and CTS(or PBS)intragastrically,and establishment of the chronic infection model was verified using polymerase chain reaction and sequencing of isolated strains.Hematoxylin and eosin staining was used to assess mucosal erosion in the stomach and toxicity to the liver and kidney.RESULTS CTS inhibited the growth of GC cells in dose-and time-dependent manners.Overexpression of CagA promoted the growth,migration,and invasion of GC cells.Importantly,we demonstrated that CTS significantly inhibited the CagAinduced abnormal proliferation,migration,and invasion of GC cells.Moreover,the expression of p-SHP2 protein in tumor tissue was related to the expression of IgG against CagA in the serum of GC patients.Additionally,CTS suppressed the protein expression levels of both SHP2 and p-SHP2 in GC cells.CTS suppressed CagA+H.pylori strain-induced mucosal erosion in the stomach of mice but had no obvious effects on the CagA-H.pylori strain group.CONCLUSION CTS inhibited CagA-induced proliferation and the epithelial-mesenchymal transition of GC cells in vitro,and CagA+H.pylori strains caused mucosal erosions of the stomach in vivo by decreasing the protein expression of SHP2.展开更多
Total tanshinones are lipophilic active constituents extracted from Salvia miltiorrhiza Bge.Tanshinone ⅡA and cryptotanshinone are the major components in total tanshinones.However, the bioavailability of both compou...Total tanshinones are lipophilic active constituents extracted from Salvia miltiorrhiza Bge.Tanshinone ⅡA and cryptotanshinone are the major components in total tanshinones.However, the bioavailability of both compounds is low due to poor water solubility. To enhance the solubility and dissolution rate of tanshinone ⅡA, cryptotanshinone and total tanshinones,three common used hydrophilic carriers including PEG 6000, poloxamer 188 and PVP K30 were used to prepare the solid dispersions at different ratios, respectively. The solid dispersions were characterised by scanning electron microscopy(SEM), differential scanning calorimetry(DSC) and Fourier transform infrared spectroscopy(FTIR). The results of powder X-ray diffraction confirmed the microcrystal state of total tanshinones in solid dispersions and no chemical interaction between total tanshinones and carriers was observed in FTIR spectra. The solubility and dissolution rate of tanshinone ⅡA and cryptotanshinone were significantly increased in all solid dispersions. Regarding tanshinone ⅡA, the solubility and dissolution rate of in solid dispersions prepared with poloxamer 188 were significantly higher than that with PEG 6000 and PVP K30. The higher solubility and dissolution rate of cryptotanshinone were obtained in solid dispersion of PVP K30 than that of PEG 6000 solid dispersions but no significant difference from poloxamer 188 solid dispersions. The results indicate that the superior carrier for preparation of tanshinone ⅡA and total tanshinones solid dispersions is poloxamer 188, and that for cryptotanshinone is PVP K30.展开更多
The clinical manifestations of variant angina is unevenly distributed during the 24 h, thusthe in vivo performance of drugs should be tailored according to the angina circadianrhythm. Cryptotanshinone(CTN) is one of t...The clinical manifestations of variant angina is unevenly distributed during the 24 h, thusthe in vivo performance of drugs should be tailored according to the angina circadianrhythm. Cryptotanshinone(CTN) is one of the representative bioactive lipid-soluble com-ponents of Danshen which has been commonly used for cardiovascular diseases such asangina pectoris. The aim of this study was to develop a novel CTN sustained-released pel-lets(CTN-SRPs) to precisely synchronize the CTN plasma concentrations with predictedoccurrence of angina pectoris for angina chronotherapy. A deconvolution-based methodwas applied to develop and optimize the CTN-SRPs. The plasma concentration-time curveof CTN immediate-released formulation after oral administration in rats was used as theweight function. The predicted plasma concentration-time curve of CTN-SRPs simulatedaccording to the incidence of variant angina during 24 h was used as the response func-tion. Then the desired drug release profile of CTN-SRPs was calculated based on deconvo-lution using weight function and response function, and subsequently used for guiding theformulation optimization. CTN-SRPs were prepared with the combinations of PVP, polox-amer 127 and EC as matrix using fluidized bed technology. An orthogonal design was em-ployed to obtain the optimal formulation with its release profile similar with the desiredone. Pharmacokinetic studies validated that the actual plasma concentration-time curve ofthese optimized CTN-SRPs was similar with the predicted one. In addition, the percent er-rors(%PE) of CTN plasma concentrations in 8–12 h were less than 10%. In conclusion, thisdeconvolution-based method could be applied to adjust the in vivo performance of drugs forangina chronotherapy.展开更多
Cryptotanshinone (CT), a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated effective in vitro antibacterial activity against all oral bacteria tested in this experiment. The antibacterial act...Cryptotanshinone (CT), a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated effective in vitro antibacterial activity against all oral bacteria tested in this experiment. The antibacterial activities of CT against oral bacteria were assessed using the checkerboard and time-kill methods to evaluate the synergistic effects of treatment with ampicillin or gentamicin. The CT was determined against oral pathogenic bacteria with MIC and MBC values ranging from 0.5 to 16 and 1 to 64 μg/mL;for am- picillin from 0.0313 to 16 and 0.125 to 32 μg/mL;for gentamicin from 2 to 256 and 4 to 512 μg/mL respectively. The range of MIC50 and MIC90 were 0.0625 - 8 μg/mL and 1 - 64 μg/mL, respectively. The combination effects of CT with antibiotics were synergistic (FIC index < 0.5) against tested oral bacteria except additive, Streptococcus sobrinus, S. criceti, and Actinobacillus actinomycetemcomitans (FIC index < 0.75 - 1.0). The MBCs were shown reducing ≥4 - 8-fold, indicating a synergistic effect as defined by a FBCI of ≤0.5. Furthermore, a time-kill study showed that the growth of the tested bacteria was completely attenuated after 3 - 6 h of treatment with the 1/2 MIC of CT, regardless of whether it was administered alone or with ampicillin or gentamicin. The results suggest that CT could be employed as a natural antibacterial agent against cariogenic and periodontopathogenic bac- teria.展开更多
[Objectives] The aim was to investigate the effect of cryptotanshinone on apoptosis of human melanoma A375 cells and its related mechanism of mitochondrial pathway.[Methods]The cytotoxic effect of cryptotanshinone on ...[Objectives] The aim was to investigate the effect of cryptotanshinone on apoptosis of human melanoma A375 cells and its related mechanism of mitochondrial pathway.[Methods]The cytotoxic effect of cryptotanshinone on apoptosis of human melanoma A375 cells was detected by MTT colorimetry.The apoptosis of melanoma A375 cells was detected with Annexin V-FITC/PI and observed by fluorescence inverted microscope.The expression of apoptosis-related proteins was detected by Western blotting.[Results]The viability of the A375 cells decreased with the increase of drug concentration.The fluorescence intensity of the cells increased with the treatment time.The expression of pro-apoptotic protein caspase-3 gradually increased,while the expression of apoptosis-inhibiting proteins p-AKT and Bcl-2 gradually reduced.[Conclusions]Cryptotanshinone induces apoptosis of human melanoma A375 cells via AKT signaling pathway,thus exerting a good cytotoxic effect on A375 cells.展开更多
Cryptotanshinone is a natural active ingredient extracted from traditional Chinese medicine Radix Salviae Miltiorrhizae.It has a variety of pharmacological effects,in addition to preventing and treating ischemic disea...Cryptotanshinone is a natural active ingredient extracted from traditional Chinese medicine Radix Salviae Miltiorrhizae.It has a variety of pharmacological effects,in addition to preventing and treating ischemic diseases,coronary artery disease Alzheimer's disease and fighting tumors.The anti-tumor effects are exerted through inhibiting tumor cell proliferation,inducing tumor cell apoptosis,inhibiting tumor metastasis and invasion,inhibiting angiogenesis and regulating reactive oxygen species levels.The studies on the anti-tumor activity and action mechanism of cryptotanshinone in recent years are reviewed in this article.展开更多
Objective:Sorafenib resistance has been a major factor limiting its clinical use as a targeted drug in liver cancer.The present study aimed to investigate whether cryptotanshinone can enhance the sensitivity of liver ...Objective:Sorafenib resistance has been a major factor limiting its clinical use as a targeted drug in liver cancer.The present study aimed to investigate whether cryptotanshinone can enhance the sensitivity of liver cancer and reduce the resistance to sorafenib.Methods:Sorafenib-resistant cells were established based on HepG2 and Huh7 cell lines.And the anti-tumor effect of sorafenib combined with cryptotanshinone on the sorafenib-resistant cells was verified by MTT,colony formation,transwell assays and tumor growth xenograft model.Moreover,the effects of the combined treatment on the expression of phosphorylated(p)-STAT3,as well as epithelial mesenchymal transition(EMT)and apoptosis related proteins of cells were evaluated by western blot analysis.Results:It was identified that cryptotanshinone inhibited the viability of both HepG2 and Huh7 cells in a dose-and time-dependent manner,and decreased p-STAT3 expression rather than total STAT3 expression at a concentration of 40μmol/L.In the sorafenib-resistant cells,sorafenib in combination with cryptotanshinone markedly inhibited cell viability,invasion and migration compared with sorafenib alone.In contrast,increased p-STAT3 level by colivelin led to the inhibition of the synergistic effect of cryptotanshinone and sorafenib not only on cell viability,but also on EMT and apoptosis,suggesting that cryptotanshinone and sorafenib may act by downregulating STAT3 signaling.Further,the inhibition of carcinogenicity effect was also verified in xenografted tumor models.Conclusion:The present results indicated that cryptotanshinone could synergize with sorafenib to inhibit the proliferative,invasive,and migratory abilities of sorafenib-resistant cells by downregulating STAT3 signaling.展开更多
Objectives The impairment of matrix metallopro- teinase-2(MMP-2)has been associated with the development of cardiac fibrosis.Although the Chinese herb Salvia miltior-rhiza has been widely used in patients with cardiov...Objectives The impairment of matrix metallopro- teinase-2(MMP-2)has been associated with the development of cardiac fibrosis.Although the Chinese herb Salvia miltior-rhiza has been widely used in patients with cardiovascular disorders,the mechanisms involved have not been elucidated. The purpose of the present study was to determine whether the administration of cryptotanshinone,an active ingredient of Salvia miltiorrhiza,could prevent the cardiac fibrosis induced by isoprenaline and to investigate the underlying mechanisms. Methods and Results Male C57BL/6 mice were submitted to receive daily injection of 0.9%saline,3 mg/kg isoprenaline, or isoprenaline plus 20 mg/kg cryptotanshinone by gastric gavage for 2 weeks.Herein,we demonstrate that cryptotanshinone can significantly ameliorate the isoprenaline-induced cardiac fibrosis,which was associated with marked up-regulation and activation of MMP-2 in ventricular myocardium. Additionally,we demonstrate that cryptotanshinone can dose-dependently upregulate and activate MMP-2 in cultured cardiac fibroblast.Moreover,incubation with cryptotanshinone also can prevent isoprenaline-induced downregulation and inactivation of MMP-2 in cultured cardiac fibroblast. Conclusions Taken together,our data suggest that cryptotanshinone may become a novel and potent antifibrotic agent. The present findings might further our understanding of the role of MMP-2 in cardiac fibrosis and antifibrotic mechanisms of cryptotanshinone.展开更多
Objective：To investigate the anti-angiogenic effect of cryptotanshinone（CPT） on human umbilical vein endothelial cells（HUVECs） and the effect of CPT on Wnt/β-catenin signaling pathway.Methods：HUVECs were incuba...Objective：To investigate the anti-angiogenic effect of cryptotanshinone（CPT） on human umbilical vein endothelial cells（HUVECs） and the effect of CPT on Wnt/β-catenin signaling pathway.Methods：HUVECs were incubated with 0,2.5,5,10,and 20 μmol/L CPT for detecting cell viability with dimethyl thiazolyl-2,5-diphenyltetrazolium bromide（MTT） assay.Then,HUVECs were incubated with 0,2.5,5,and 10 μmol/L CPT for detecting endothelial cell migration,invasion,and tubular-like structure formation with wound healing,transwell invasion and matrigel tube formation assays,respectively.To gain insight into CPT-mediated signaling,the effects of CPT on T-cell factor/lymphocyte enhancer factor（TCF/LEF） transcription factors were detected by the Dual-luciferase reporter assay.Next,the nuclear expression of p-catenin was evaluated using Western blot and immunochemistry.Finally,vascular endothelial growth factor（VEGF） and cyclin D1,downstream proteins of the Wnt pathway were examined with Western blot.Results：CPT dose-dependently suppressed endothelial cell viability,migration,invasion,and tubular-like structure formation.In particular,CPT blocked β-catenindependent transcription in HUVECs in a dose-dependent manner.In Western bolt,10 μ mol/L CPT decreased expression of β-catenin in nucleus of HUVECs（P〈0.01）.In immunohistochemistry,β-catenin was more potent in response to LiCI（an activator of the pathway） treatment.However,the signals were weaker in the nucleus of the CPT（10 μmol/L） group,compared to the positive control.Also,VEGF and cyclin D1 were both eliminated by CPT in 5 and 10 μ mol/L doses（P〈0.05）.Conclosion：Our study supported the role of CPT as an angiogenic inhibitor,which may impact on the Wnt/β-catenin signaling pathway.展开更多
Anticancer targets of cryptotanshinone were evaluated and rapidly forecasted with PharmMapper, a reverse pharmacophore-based screening platform, as well as drug target databases, including PDTD, DrugBank and TTD. The ...Anticancer targets of cryptotanshinone were evaluated and rapidly forecasted with PharmMapper, a reverse pharmacophore-based screening platform, as well as drug target databases, including PDTD, DrugBank and TTD. The pathway analyses for the collection of anticancer targets screened were carried out based on the KEGG pathway database, followed by the forecast of potential pharmacological activities and pathways of the effects of cryptotanshinone, and verification of some of the targets screened using whole cell tests. The results showed that a total of eight targets with anticancer potential were screened, including MAP2K1, RARα, RXRα, PDK1, CHK1, AR, Ang-1 R, and Kif11. These targets are mainly related to four aspects of the cancer growth: the cell cycle, angiogenesis, apoptosis, and androgen receptor. The cell tests showed that cryptotanshinone can inhibit the viability of human hepatoma cells SMMC-7721, which is related to the reduction of expression of MAP2K1 mRNA. This method provides a strong clue for the study of the anticancer effects and mechanisms of action of cryptotanshinone in the future.展开更多
Background Cryptotanshinone （CT） was originally isolated from the dried roots of Salvia militorrhiza, an herb that is used extensively in Asian medicine and the extracts of this herb have been used in the treatment ...Background Cryptotanshinone （CT） was originally isolated from the dried roots of Salvia militorrhiza, an herb that is used extensively in Asian medicine and the extracts of this herb have been used in the treatment of several pathologies, including cardiovascular diseases, hematological abnormalities, hepatitis, and hyperlipidemia, but no studies had been carried on the treatment for rheumatic diseases with it. This study aimed to investigate the effects of cryptotanshinone on immune functions in rats with adjuvant arthritis （AA）. Methods Complete Freund＇s adjuvant was used to induce AA in rats. Thymus and spleen was aseptically taken from normal rats and the AA rats. Then a thymus lymphoid cell suspension, splenic lymphoid cell suspension and peritoneal macrophage cell suspension were prepared. After adding CT （0.1 ug/ml, 1.0 ug/ml, 10 ug/ml, 100 ug/ml, 1000 ug/ml） into the suspension, T and B lymphocytes proliferation was determined by 3-（4,5-2 dimethylthiazal-2yl）2,5- diphenyltetrazoliumbromide （MTT） assay. And the activities of interleukin-1 （IL-1） and IL-2 were measured by the mouse lymphocytes proliferation assay. Results Thymic T and splenic B lymphocyte proliferation of the AA rat was significantly lower, and could be stored through using CT in vitro. CT （100ug/ml and 1000ug/ml） increased T or B lymphocytes proliferation in vitro （P 〈0.01）. In AA rats, the levels of IL-1 released by abdominal PMφ significantly increased whereas the level of IL-2 released by T cells decreased in vitro. CT （1000 pg/ml） decreased the production of IL-1 and promoted production of IL-2 in vitro （P 〈0.05）. Conclusions CT can ameliorate the abnormal immunological functions in AA rats.展开更多
Background Cryptotanshinone （CT） is the major active constituent of Salvia miltiorrhiza Bunge. The present study was carried out to investigate the effects of CT on rats with adjuvant arthritis （AA）. Methods AA wa...Background Cryptotanshinone （CT） is the major active constituent of Salvia miltiorrhiza Bunge. The present study was carried out to investigate the effects of CT on rats with adjuvant arthritis （AA）. Methods AA was induced by the metatarsal footpad injection with complete Freund＇s adjuvant in male Sprague-Dawley rats. The secondary inflammatory reaction was evaluated by hind paw swelling and the polyarthritis index. Activity of interleukin-1 （IL-1） was detected by the concanavalin A-induced thymocytes proliferation assay. The lymphocytes proliferation and IL-2 production were assayed by 3-（4,5-2dimethylthiazal-2yl）2,5-diphenyltetrazoliumbromide （MTT） and activated mouse splenocytes proliferation, respectively. Results Intragastric administration of CT （50 and 100 mg/kg） significantly decreased secondary inflammatory reactions and increased the spleen and thymus index. There was a marked immunologic and inflammatory response in the AA model, which was accompanied by the decrease of thymocyte proliferation and IL-2 production as well as the increase of IL-1 production. CT apparently enhanced thymocyte proliferation and decreased IL-1 production in AA rats. Conclusion These results indicate that CT may exert its anti-inflammatory and immunoregulatory effects through inhibiting lymphocyte proliferation and production of pro-inflammatory mediators.展开更多
The reactions of cryptotanshinone and tanshinone IIA with cadaverine and putrescine were investigated. Six new compounds, four with imidazole functional groups and two with oxazole groups, were obtained. The possibl...The reactions of cryptotanshinone and tanshinone IIA with cadaverine and putrescine were investigated. Six new compounds, four with imidazole functional groups and two with oxazole groups, were obtained. The possible reaction mechanism was proposed.展开更多
The reaction of cryptotanshinone and tanshinone IIA with several biogenic amine metabolites involved in the pathogenic pathways of HE were investigated and eight 1,2,3,4- tetrahydrophenanthrene derivatives, 2-6 and 8-...The reaction of cryptotanshinone and tanshinone IIA with several biogenic amine metabolites involved in the pathogenic pathways of HE were investigated and eight 1,2,3,4- tetrahydrophenanthrene derivatives, 2-6 and 8-10, were obtained. The probable mechanism on reaction was discussed.展开更多
Objective A reversed-phase HPLC method was established for the simultaneous determination of five hydrophilic and lipophilic components in the roots of Salvia miltiorrhiza. Methods Hydrophilic components including dan...Objective A reversed-phase HPLC method was established for the simultaneous determination of five hydrophilic and lipophilic components in the roots of Salvia miltiorrhiza. Methods Hydrophilic components including danshensu, protocatechuic aldehyde, and salvianolic acid B, and lipophilic components such as cryptotanshinone and tanshinone liA, were successfully separated on a Waters Symmetry C18 reverse- phase column （250 mm×4.6 mm, 5 μm）, with acetonitrile-0.5% phosphoric acid （gradient elution） as mobile phase, the detection wavelength was set at 281 nm with flow rate of 1.0 mL/min, and the column temperature was maintained at 30 ~C. Results The recovery of the method was in the range of 95.1%-102.5% and the precision was less than 3% for all five analytes. All the compounds showed good linearity （R2 〉 0.9990） in a relatively wide concentration range. Therefore, this HPLC method demonstrated good reproducibility, stability, and accuracy in validation studies. Conclusion Simultaneous quantification of the multiple components by HPLC would be a better strategy for the quality evaluation on the roots of S. miltiorrhiza.展开更多
基金Science and Technology Plan of Hainan Province(Clinical Research Center),No.LCYX202103 and No.LCYX202204Hainan Province Science and Technology Special Fund,No.ZDYF2022SHFZ067Hainan Province Clinical Medical Center.
文摘BACKGROUND Cryptotanshinone(CPT)has wide biological functions,including anti-oxidative,antifibrosis,and anti-inflammatory properties.However,the effect of CPT on hepatic fibrosis is unknown.AIM To investigate the effects of CPT treatment on hepatic fibrosis and its underlying mechanism of action.METHODS Hepatic stellate cells(HSCs)and normal hepatocytes were treated with different concentrations of CPT and salubrinal.The CCK-8 assay was used to determine cell viability.Flow cytometry was used to measure apoptosis and cell cycle arrest.Reverse transcription polymerase chain reaction(RT-PCR)and Western blot analyses were used to measure mRNA levels and protein expression of endoplasmic reticulum stress(ERS)signaling pathway related molecules,respectively.Carbon tetrachloride(CCL4)was used to induce in vivo hepatic fibrosis in mice.Mice were treated with CPT and salubrinal,and blood and liver samples were collected for histopathological examination.RESULTS We found that CPT treatment significantly reduced fibrogenesis by modulating the synthesis and degradation of the extracellular matrix in vitro.CPT inhibited cell proliferation and induced cell cycle arrest at the G2/M phase in cultured HSCs.Furthermore,we found that CPT promoted apoptosis of activated HSCs by upregulating expression of ERS markers(CHOP and GRP78)and activating ERS pathway molecules(PERK,IRE1α,and ATF4),which were inhibited by salubrinal.Inhibition of ERS by salubrinal partially eliminated the therapeutic effect of CPT in our CCL4-induced hepatic fibrosis mouse model.CONCLUSION CPT can promote apoptosis of HSCs and alleviate hepatic fibrosis through modulating the ERS pathway,which represents a promising strategy for treating hepatic fibrosis.
基金funded by the Indian Council of Medical Research with Grant Number 2020-8817,New Delhi,India。
文摘Objective:To evaluate the neuroprotective effect of cryptotanshinone against cladribine-induced cognitive impairment in rats.Methods:Rats were administered with cladribine(1 mg/kg,p.o.)and cryptotanshinone(10 and 20 mg/kg,i.p.)for four weeks.Behavioral tests such as Morris water maze and elevated plus maze were conducted to check memory impairment caused by cladribine.On day 29,all rats were sacrificed,and the brains were separated for estimation of neuroinflammatory factors,biochemical parameters,neurotransmitters,Aβ(1-42),blood-brain barrier permeability,nuclear factor erythroid 2-related factor 2(Nrf2),and brain-derived neurotrophic factor(BDNF).Results:Treatment with cryptotanshinone dose-dependently enhanced spatial memory,improved the levels of neurotransmitter and antioxidant enzymes,and suppressed proinflammatory cytokine release.Cryptotanshinone also decreased Aβ(1-42)accumulation and increased the levels of Nrf2 and BDNF in the hippocampus.Additionally,the histopathological results showed that cryptotanshinone reduced cladribine-induced neuronal death in the hippocampus.Conclusions:Cryptotanshinone exhibits a promising neuroprotective effect against cladribine-induced cognitive impairment in preclinical studies,and may be a potential phytochemical for the treatment and management of cognitive impairment.
基金National Natural Science Foundation of China,No.81572350。
文摘BACKGROUND Approximately 90%of new cases of noncardiac gastric cancer(GC)are related to Helicobacter pylori(H.pylori),and cytotoxin-associated gene A(CagA)is one of the main pathogenic factors.Recent studies have shown that the pharmacological effects of cryptotanshinone(CTS)can be used to treat a variety of tumors.However,the effects of CTS on H.pylori,especially CagA+strain-induced gastric mucosal lesions,on the development of GC is unknown.AIM To assess the role of CTS in CagA-induced proliferation and metastasis of GC cells,and determine if CagA+H.pylori strains causes pathological changes in the gastric mucosa of mice.METHODS The effects of CTS on the proliferation of GC cells were assessed using the Cell Counting Kit-8(CCK-8)assay,and the abnormal growth,migration and invasion caused by CagA were detected by CCK-8 and transwell assays.After transfection with pSR-HA-CagA and treatment with CTS,proliferation and metastasis were evaluated by CCK-8 and transwell assays,respectively,and the expression of Src homology 2(SH2)domain–containing phosphatase 2(SHP2)and phosphorylated SHP2(p-SHP2)was detected using western blotting in AGS cells.The enzymelinked immunosorbent assay was used to determine the immunoglobulin G(IgG)level against CagA in patient serum.Mice were divided into four groups and administered H.pylori strains(CagA+or CagA-)and CTS(or PBS)intragastrically,and establishment of the chronic infection model was verified using polymerase chain reaction and sequencing of isolated strains.Hematoxylin and eosin staining was used to assess mucosal erosion in the stomach and toxicity to the liver and kidney.RESULTS CTS inhibited the growth of GC cells in dose-and time-dependent manners.Overexpression of CagA promoted the growth,migration,and invasion of GC cells.Importantly,we demonstrated that CTS significantly inhibited the CagAinduced abnormal proliferation,migration,and invasion of GC cells.Moreover,the expression of p-SHP2 protein in tumor tissue was related to the expression of IgG against CagA in the serum of GC patients.Additionally,CTS suppressed the protein expression levels of both SHP2 and p-SHP2 in GC cells.CTS suppressed CagA+H.pylori strain-induced mucosal erosion in the stomach of mice but had no obvious effects on the CagA-H.pylori strain group.CONCLUSION CTS inhibited CagA-induced proliferation and the epithelial-mesenchymal transition of GC cells in vitro,and CagA+H.pylori strains caused mucosal erosions of the stomach in vivo by decreasing the protein expression of SHP2.
文摘Total tanshinones are lipophilic active constituents extracted from Salvia miltiorrhiza Bge.Tanshinone ⅡA and cryptotanshinone are the major components in total tanshinones.However, the bioavailability of both compounds is low due to poor water solubility. To enhance the solubility and dissolution rate of tanshinone ⅡA, cryptotanshinone and total tanshinones,three common used hydrophilic carriers including PEG 6000, poloxamer 188 and PVP K30 were used to prepare the solid dispersions at different ratios, respectively. The solid dispersions were characterised by scanning electron microscopy(SEM), differential scanning calorimetry(DSC) and Fourier transform infrared spectroscopy(FTIR). The results of powder X-ray diffraction confirmed the microcrystal state of total tanshinones in solid dispersions and no chemical interaction between total tanshinones and carriers was observed in FTIR spectra. The solubility and dissolution rate of tanshinone ⅡA and cryptotanshinone were significantly increased in all solid dispersions. Regarding tanshinone ⅡA, the solubility and dissolution rate of in solid dispersions prepared with poloxamer 188 were significantly higher than that with PEG 6000 and PVP K30. The higher solubility and dissolution rate of cryptotanshinone were obtained in solid dispersion of PVP K30 than that of PEG 6000 solid dispersions but no significant difference from poloxamer 188 solid dispersions. The results indicate that the superior carrier for preparation of tanshinone ⅡA and total tanshinones solid dispersions is poloxamer 188, and that for cryptotanshinone is PVP K30.
基金supported by the National Natural Science Foundation of China (No.81473151)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘The clinical manifestations of variant angina is unevenly distributed during the 24 h, thusthe in vivo performance of drugs should be tailored according to the angina circadianrhythm. Cryptotanshinone(CTN) is one of the representative bioactive lipid-soluble com-ponents of Danshen which has been commonly used for cardiovascular diseases such asangina pectoris. The aim of this study was to develop a novel CTN sustained-released pel-lets(CTN-SRPs) to precisely synchronize the CTN plasma concentrations with predictedoccurrence of angina pectoris for angina chronotherapy. A deconvolution-based methodwas applied to develop and optimize the CTN-SRPs. The plasma concentration-time curveof CTN immediate-released formulation after oral administration in rats was used as theweight function. The predicted plasma concentration-time curve of CTN-SRPs simulatedaccording to the incidence of variant angina during 24 h was used as the response func-tion. Then the desired drug release profile of CTN-SRPs was calculated based on deconvo-lution using weight function and response function, and subsequently used for guiding theformulation optimization. CTN-SRPs were prepared with the combinations of PVP, polox-amer 127 and EC as matrix using fluidized bed technology. An orthogonal design was em-ployed to obtain the optimal formulation with its release profile similar with the desiredone. Pharmacokinetic studies validated that the actual plasma concentration-time curve ofthese optimized CTN-SRPs was similar with the predicted one. In addition, the percent er-rors(%PE) of CTN plasma concentrations in 8–12 h were less than 10%. In conclusion, thisdeconvolution-based method could be applied to adjust the in vivo performance of drugs forangina chronotherapy.
文摘Cryptotanshinone (CT), a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated effective in vitro antibacterial activity against all oral bacteria tested in this experiment. The antibacterial activities of CT against oral bacteria were assessed using the checkerboard and time-kill methods to evaluate the synergistic effects of treatment with ampicillin or gentamicin. The CT was determined against oral pathogenic bacteria with MIC and MBC values ranging from 0.5 to 16 and 1 to 64 μg/mL;for am- picillin from 0.0313 to 16 and 0.125 to 32 μg/mL;for gentamicin from 2 to 256 and 4 to 512 μg/mL respectively. The range of MIC50 and MIC90 were 0.0625 - 8 μg/mL and 1 - 64 μg/mL, respectively. The combination effects of CT with antibiotics were synergistic (FIC index < 0.5) against tested oral bacteria except additive, Streptococcus sobrinus, S. criceti, and Actinobacillus actinomycetemcomitans (FIC index < 0.75 - 1.0). The MBCs were shown reducing ≥4 - 8-fold, indicating a synergistic effect as defined by a FBCI of ≤0.5. Furthermore, a time-kill study showed that the growth of the tested bacteria was completely attenuated after 3 - 6 h of treatment with the 1/2 MIC of CT, regardless of whether it was administered alone or with ampicillin or gentamicin. The results suggest that CT could be employed as a natural antibacterial agent against cariogenic and periodontopathogenic bac- teria.
基金Supported by Multigrain Production and Processing Characteristic Discipline Construction ProjectPostdoctoral Scientific Research Foundation of Heilongjiang Province of China(LBH-Q13132)
文摘[Objectives] The aim was to investigate the effect of cryptotanshinone on apoptosis of human melanoma A375 cells and its related mechanism of mitochondrial pathway.[Methods]The cytotoxic effect of cryptotanshinone on apoptosis of human melanoma A375 cells was detected by MTT colorimetry.The apoptosis of melanoma A375 cells was detected with Annexin V-FITC/PI and observed by fluorescence inverted microscope.The expression of apoptosis-related proteins was detected by Western blotting.[Results]The viability of the A375 cells decreased with the increase of drug concentration.The fluorescence intensity of the cells increased with the treatment time.The expression of pro-apoptotic protein caspase-3 gradually increased,while the expression of apoptosis-inhibiting proteins p-AKT and Bcl-2 gradually reduced.[Conclusions]Cryptotanshinone induces apoptosis of human melanoma A375 cells via AKT signaling pathway,thus exerting a good cytotoxic effect on A375 cells.
基金Supported by Multigrain Production and Processing Characteristic Discipline Construction ProjectPostdoctoral Scientific Research Foundation of Heilongjiang Province of China(LBH-Q13132)
文摘Cryptotanshinone is a natural active ingredient extracted from traditional Chinese medicine Radix Salviae Miltiorrhizae.It has a variety of pharmacological effects,in addition to preventing and treating ischemic diseases,coronary artery disease Alzheimer's disease and fighting tumors.The anti-tumor effects are exerted through inhibiting tumor cell proliferation,inducing tumor cell apoptosis,inhibiting tumor metastasis and invasion,inhibiting angiogenesis and regulating reactive oxygen species levels.The studies on the anti-tumor activity and action mechanism of cryptotanshinone in recent years are reviewed in this article.
文摘Objective:Sorafenib resistance has been a major factor limiting its clinical use as a targeted drug in liver cancer.The present study aimed to investigate whether cryptotanshinone can enhance the sensitivity of liver cancer and reduce the resistance to sorafenib.Methods:Sorafenib-resistant cells were established based on HepG2 and Huh7 cell lines.And the anti-tumor effect of sorafenib combined with cryptotanshinone on the sorafenib-resistant cells was verified by MTT,colony formation,transwell assays and tumor growth xenograft model.Moreover,the effects of the combined treatment on the expression of phosphorylated(p)-STAT3,as well as epithelial mesenchymal transition(EMT)and apoptosis related proteins of cells were evaluated by western blot analysis.Results:It was identified that cryptotanshinone inhibited the viability of both HepG2 and Huh7 cells in a dose-and time-dependent manner,and decreased p-STAT3 expression rather than total STAT3 expression at a concentration of 40μmol/L.In the sorafenib-resistant cells,sorafenib in combination with cryptotanshinone markedly inhibited cell viability,invasion and migration compared with sorafenib alone.In contrast,increased p-STAT3 level by colivelin led to the inhibition of the synergistic effect of cryptotanshinone and sorafenib not only on cell viability,but also on EMT and apoptosis,suggesting that cryptotanshinone and sorafenib may act by downregulating STAT3 signaling.Further,the inhibition of carcinogenicity effect was also verified in xenografted tumor models.Conclusion:The present results indicated that cryptotanshinone could synergize with sorafenib to inhibit the proliferative,invasive,and migratory abilities of sorafenib-resistant cells by downregulating STAT3 signaling.
文摘Objectives The impairment of matrix metallopro- teinase-2(MMP-2)has been associated with the development of cardiac fibrosis.Although the Chinese herb Salvia miltior-rhiza has been widely used in patients with cardiovascular disorders,the mechanisms involved have not been elucidated. The purpose of the present study was to determine whether the administration of cryptotanshinone,an active ingredient of Salvia miltiorrhiza,could prevent the cardiac fibrosis induced by isoprenaline and to investigate the underlying mechanisms. Methods and Results Male C57BL/6 mice were submitted to receive daily injection of 0.9%saline,3 mg/kg isoprenaline, or isoprenaline plus 20 mg/kg cryptotanshinone by gastric gavage for 2 weeks.Herein,we demonstrate that cryptotanshinone can significantly ameliorate the isoprenaline-induced cardiac fibrosis,which was associated with marked up-regulation and activation of MMP-2 in ventricular myocardium. Additionally,we demonstrate that cryptotanshinone can dose-dependently upregulate and activate MMP-2 in cultured cardiac fibroblast.Moreover,incubation with cryptotanshinone also can prevent isoprenaline-induced downregulation and inactivation of MMP-2 in cultured cardiac fibroblast. Conclusions Taken together,our data suggest that cryptotanshinone may become a novel and potent antifibrotic agent. The present findings might further our understanding of the role of MMP-2 in cardiac fibrosis and antifibrotic mechanisms of cryptotanshinone.
基金Supported by National Natural Science Foundation of China(No.81170270)Medicine and Technology Program of Zhejiang Province(No.2013KYB188)
文摘Objective：To investigate the anti-angiogenic effect of cryptotanshinone（CPT） on human umbilical vein endothelial cells（HUVECs） and the effect of CPT on Wnt/β-catenin signaling pathway.Methods：HUVECs were incubated with 0,2.5,5,10,and 20 μmol/L CPT for detecting cell viability with dimethyl thiazolyl-2,5-diphenyltetrazolium bromide（MTT） assay.Then,HUVECs were incubated with 0,2.5,5,and 10 μmol/L CPT for detecting endothelial cell migration,invasion,and tubular-like structure formation with wound healing,transwell invasion and matrigel tube formation assays,respectively.To gain insight into CPT-mediated signaling,the effects of CPT on T-cell factor/lymphocyte enhancer factor（TCF/LEF） transcription factors were detected by the Dual-luciferase reporter assay.Next,the nuclear expression of p-catenin was evaluated using Western blot and immunochemistry.Finally,vascular endothelial growth factor（VEGF） and cyclin D1,downstream proteins of the Wnt pathway were examined with Western blot.Results：CPT dose-dependently suppressed endothelial cell viability,migration,invasion,and tubular-like structure formation.In particular,CPT blocked β-catenindependent transcription in HUVECs in a dose-dependent manner.In Western bolt,10 μ mol/L CPT decreased expression of β-catenin in nucleus of HUVECs（P〈0.01）.In immunohistochemistry,β-catenin was more potent in response to LiCI（an activator of the pathway） treatment.However,the signals were weaker in the nucleus of the CPT（10 μmol/L） group,compared to the positive control.Also,VEGF and cyclin D1 were both eliminated by CPT in 5 and 10 μ mol/L doses（P〈0.05）.Conclosion：Our study supported the role of CPT as an angiogenic inhibitor,which may impact on the Wnt/β-catenin signaling pathway.
基金supported by the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(No.11KJB360004)the Jiangsu Provincial Natural Science Foundation of China(No.BK2012458),the National Natural Science Foundation of China(Nos.81373232,81173174,81270514)+1 种基金the National Key Technology R&D Program during the 11th Five-Year Plan Period(No.2008BAI51B02)the Doctoral Fund of Ministry of Education of China(No.20113237110008)
文摘Anticancer targets of cryptotanshinone were evaluated and rapidly forecasted with PharmMapper, a reverse pharmacophore-based screening platform, as well as drug target databases, including PDTD, DrugBank and TTD. The pathway analyses for the collection of anticancer targets screened were carried out based on the KEGG pathway database, followed by the forecast of potential pharmacological activities and pathways of the effects of cryptotanshinone, and verification of some of the targets screened using whole cell tests. The results showed that a total of eight targets with anticancer potential were screened, including MAP2K1, RARα, RXRα, PDK1, CHK1, AR, Ang-1 R, and Kif11. These targets are mainly related to four aspects of the cancer growth: the cell cycle, angiogenesis, apoptosis, and androgen receptor. The cell tests showed that cryptotanshinone can inhibit the viability of human hepatoma cells SMMC-7721, which is related to the reduction of expression of MAP2K1 mRNA. This method provides a strong clue for the study of the anticancer effects and mechanisms of action of cryptotanshinone in the future.
文摘Background Cryptotanshinone （CT） was originally isolated from the dried roots of Salvia militorrhiza, an herb that is used extensively in Asian medicine and the extracts of this herb have been used in the treatment of several pathologies, including cardiovascular diseases, hematological abnormalities, hepatitis, and hyperlipidemia, but no studies had been carried on the treatment for rheumatic diseases with it. This study aimed to investigate the effects of cryptotanshinone on immune functions in rats with adjuvant arthritis （AA）. Methods Complete Freund＇s adjuvant was used to induce AA in rats. Thymus and spleen was aseptically taken from normal rats and the AA rats. Then a thymus lymphoid cell suspension, splenic lymphoid cell suspension and peritoneal macrophage cell suspension were prepared. After adding CT （0.1 ug/ml, 1.0 ug/ml, 10 ug/ml, 100 ug/ml, 1000 ug/ml） into the suspension, T and B lymphocytes proliferation was determined by 3-（4,5-2 dimethylthiazal-2yl）2,5- diphenyltetrazoliumbromide （MTT） assay. And the activities of interleukin-1 （IL-1） and IL-2 were measured by the mouse lymphocytes proliferation assay. Results Thymic T and splenic B lymphocyte proliferation of the AA rat was significantly lower, and could be stored through using CT in vitro. CT （100ug/ml and 1000ug/ml） increased T or B lymphocytes proliferation in vitro （P 〈0.01）. In AA rats, the levels of IL-1 released by abdominal PMφ significantly increased whereas the level of IL-2 released by T cells decreased in vitro. CT （1000 pg/ml） decreased the production of IL-1 and promoted production of IL-2 in vitro （P 〈0.05）. Conclusions CT can ameliorate the abnormal immunological functions in AA rats.
文摘Background Cryptotanshinone （CT） is the major active constituent of Salvia miltiorrhiza Bunge. The present study was carried out to investigate the effects of CT on rats with adjuvant arthritis （AA）. Methods AA was induced by the metatarsal footpad injection with complete Freund＇s adjuvant in male Sprague-Dawley rats. The secondary inflammatory reaction was evaluated by hind paw swelling and the polyarthritis index. Activity of interleukin-1 （IL-1） was detected by the concanavalin A-induced thymocytes proliferation assay. The lymphocytes proliferation and IL-2 production were assayed by 3-（4,5-2dimethylthiazal-2yl）2,5-diphenyltetrazoliumbromide （MTT） and activated mouse splenocytes proliferation, respectively. Results Intragastric administration of CT （50 and 100 mg/kg） significantly decreased secondary inflammatory reactions and increased the spleen and thymus index. There was a marked immunologic and inflammatory response in the AA model, which was accompanied by the decrease of thymocyte proliferation and IL-2 production as well as the increase of IL-1 production. CT apparently enhanced thymocyte proliferation and decreased IL-1 production in AA rats. Conclusion These results indicate that CT may exert its anti-inflammatory and immunoregulatory effects through inhibiting lymphocyte proliferation and production of pro-inflammatory mediators.
基金the Guangzhou City Science Foundation(2002 Z1-E5011)Guangdong Provincial Science Foundation of China(2003 C104014)for the financial support.
文摘The reactions of cryptotanshinone and tanshinone IIA with cadaverine and putrescine were investigated. Six new compounds, four with imidazole functional groups and two with oxazole groups, were obtained. The possible reaction mechanism was proposed.
基金The program is sponsored by the Guangzhou City Science Foundation(2000-Z-021-01)Guangdong Provincial Science Foundation(2KM04103S).
文摘The reaction of cryptotanshinone and tanshinone IIA with several biogenic amine metabolites involved in the pathogenic pathways of HE were investigated and eight 1,2,3,4- tetrahydrophenanthrene derivatives, 2-6 and 8-10, were obtained. The probable mechanism on reaction was discussed.
基金National Department Public Benefit Research Foundation(201107011)
文摘Objective A reversed-phase HPLC method was established for the simultaneous determination of five hydrophilic and lipophilic components in the roots of Salvia miltiorrhiza. Methods Hydrophilic components including danshensu, protocatechuic aldehyde, and salvianolic acid B, and lipophilic components such as cryptotanshinone and tanshinone liA, were successfully separated on a Waters Symmetry C18 reverse- phase column （250 mm×4.6 mm, 5 μm）, with acetonitrile-0.5% phosphoric acid （gradient elution） as mobile phase, the detection wavelength was set at 281 nm with flow rate of 1.0 mL/min, and the column temperature was maintained at 30 ~C. Results The recovery of the method was in the range of 95.1%-102.5% and the precision was less than 3% for all five analytes. All the compounds showed good linearity （R2 〉 0.9990） in a relatively wide concentration range. Therefore, this HPLC method demonstrated good reproducibility, stability, and accuracy in validation studies. Conclusion Simultaneous quantification of the multiple components by HPLC would be a better strategy for the quality evaluation on the roots of S. miltiorrhiza.
