Objective： Ce/astrus orbicu/atus Thunb. has been used for thousands of years in China as a remedy against cancer and inflammatory diseases. This study aims to investigate whether C. orbiculatus extract （COE） could ...Objective： Ce/astrus orbicu/atus Thunb. has been used for thousands of years in China as a remedy against cancer and inflammatory diseases. This study aims to investigate whether C. orbiculatus extract （COE） could inhibit angiogenesis, which is the pivotal step in tumor growth, invasiveness, and metastasis. Methods： In this study, the extract from the stem of C. orbiculatus was used. Mouse hepatic carcinoma cells （Hepat-6） were treated with COE in different nontoxic concentrations （10, 20, 40, 80, and 160 μg/mL）. The mRNA and protein expression levels of vascular endothelial growth factor （VEGF） were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot, respectively; the active fractions were further tested on C57BL/6 mice and human umbilical vein endothelial cells （HUVEC） for any antiangiogenic effects. Results： COE significantly inhibited proliferation and induced apoptosis in Hepal-6 cells and inhibited VEGF expression at both mRNA and protein levels. Furthermore, this agent inhibited the formation of the capillary-like structure in primary cultured HUVEC in a dose-dependent manner. In vivo, COE significantly reduced the volume and weight of solid tumors with low adverse effects and decreased tumor angiogenesis. Conclusions： In summary, COE could be used to treat hepatic carcinoma. The mechanisms of the antitumor activity of COE may be due to its effects against tumor angiogenesis by targeting the VEGF protein.展开更多
Objective: To evaluate the effects of Celastrus Orbiculatus extracts(COE) on metastasis in hypoxiainduced hepatocellular carcinoma cells(Hep G2) and to explore the underlying molecular mechanisms. Methods:The effect o...Objective: To evaluate the effects of Celastrus Orbiculatus extracts(COE) on metastasis in hypoxiainduced hepatocellular carcinoma cells(Hep G2) and to explore the underlying molecular mechanisms. Methods:The effect of COE(160, 200 and 240 μg/mL) on cell viability, scratch-wound, invasion and migration were studied by 3-4,5-dimethyl-2-thiazolyl-2,5-diphenyl-2-H-tetrazolium bromide(MTT), scratch-wound and transwell assays, respectively. Co Cl2 was used to establish a hypoxia model in vitro. Effects of COE on the expressions of E-cadherin, vimentin and N-cadherin were investigated with Western blot and immuno?uorescence analysis,respectively. Results: COE inhibited proliferation and metastasis of hypoxia-induced hepatocellular carcinoma cells in a dose-dependent manner(P<0.01). Furthermore, the expression of epithelial-mesenchymal transition(EMT) related markers were also remarkably suppressed in a dose-dependent manner(P<0.01). In addition, the upstream signaling pathways, including the hypoxia-inducible factor 1α(Hif-1α) and Twist1 were suppressed by COE. Additionally, the Hif-1α inhibitor 3-5'-hydroxymethyl-2'-furyl)-1-benzylindazole(YC-1), potently suppressed cell invasion and migration as well as expression of EMT in hypoxia-induced Hep G2 cells. Similarly, the combined treatment with COE and YC-1 showed a synergistic effect(P<0.01) compared with the treatment with COE or YC-1 alone in hypoxia-induced Hep G2 cells. Conclusions: COE signi?cantly inhibited the tumor metastasis and EMT by suppressing Hif-1α/Twist1 signaling pathway in hypoxia-induced Hep G2 cell. Thus, COE might have potential effect to inhibit the progression of Hep G2 in the context of tumor hypoxia.展开更多
Objective To discuss the antitumor mechanism preliminarily by observing effects of Celastrus orbiculatus extracts(COE) on vascular endothelial growth factor(VEGF) mRNA and protein expression in hepatoma(Hepa1-6) cells...Objective To discuss the antitumor mechanism preliminarily by observing effects of Celastrus orbiculatus extracts(COE) on vascular endothelial growth factor(VEGF) mRNA and protein expression in hepatoma(Hepa1-6) cells of mice.Methods Hepa1-6 cells were treated with COE at different nontoxic concentration(0,10,20,40,80,and 160 μg/mL) for 16 h.The mRNA and protein expressions of VEGF were detected by reverse transcription-PCR and Western Blotting,respectively.Results COE significantly inhibited VEGF expression at both mRNA and protein levels in a dose-dependent manner.Conclusion COE can inhibit VEGF expression in Hepa1-6 cells,therefore suggest that VEGF could be chosen as an therapeutic target for COE in the context of cancer chemoprevention and anticancer therapy.展开更多
Objective:To characterize the molecular mechanism underlying the antineoplastic activity of Celastrus orbiculatus Thunb.extracts(COE).Methods:The human hepatocellular carcinoma HepG2 cells with mammalian target of rap...Objective:To characterize the molecular mechanism underlying the antineoplastic activity of Celastrus orbiculatus Thunb.extracts(COE).Methods:The human hepatocellular carcinoma HepG2 cells with mammalian target of rapamycin(mTOR)knockdown expressed(HepG2/mTOR-)were constructed using molecular biological technology.In vitro,the HepG2/mTOR-cells were treated with COE at various concentrations(10,20,40,80,160 and 320μg/mL).Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assays.According to the half-maximal inhibitory concentration(IC50)value(140 mg/L),the concentrations of COE in the subsequent experiment was set to alleviate cytotoxicity.The HepG2/mTOR-cells were divided into 5 groups:negative control(untreated),COE treatment groups(40,80,120 mg/L COE)and positive control group(cisplatin,DDP,2 mg/L),respectively.Wild-type HepG2 cells were used as a blank control.The effects of COE on the cell apoptosis were analyzed by flow cytometry and transmission electronic microscopy(TEM),respectively.The protein expression levels of mTOR signal pathways were determined by Western blotting.In vivo,HepG2/mTOR-cells(2×106 cell/mice)were subcutaneously injected into the right flank of nude mice.Thirty-six female nude mice were randomly assigned to 6 groups according to body weight(6 mice per group)as follows:solvent vehicle control,Banmao Capsule treated group(BM,195 mg/kg),Tegafur,Gimeracil and Oteracil Potassium Capsules(10 mg/kg)treated group,and different dosages of COE(10,20,40 mg/kg)groups.Tumor growth was monitored and immunohistochemical staining was used to examine the expression of apoptosis-related proteins in tumor tissues.Results:COE inhibited the proliferation significantly in a concentration-dependent manner in HepG2/mTOR-cells(P<0.01).COE significantly induced the apoptosis of HepG2/mTOR-cells(P<0.01),and the apoptotic bodies can be observed under TEM.COE significantly inhibits the proteins expression of mTORrelated signal pathways.In vivo,COE significantly in展开更多
Objective To examine the immunoregulation of Celastrus orbiculatus extracts (COE), a traditional Chinese medicine, on maturation and function of dendritic cells (DCs) in vitro and in vivo. Methods In vitro, after trea...Objective To examine the immunoregulation of Celastrus orbiculatus extracts (COE), a traditional Chinese medicine, on maturation and function of dendritic cells (DCs) in vitro and in vivo. Methods In vitro, after treated with COE in different nontoxic concentrations (0, 10, 20, 40, 80, and 160 μg/mL) for 5 d, the surface immunological molecules and cytokine secretion of mice bone marrow-derived DCs in response to COE were analyzed by flow cytometric analysis (FACS) and enzyme linked immunosorbent assay (ELISA), respectively. In vivo, mouse hepatoma cells (Hepa1-6, 1 × 106) were injected sc and were treated with different dosages of COE (10, 20 or 40 mg/kg/d). Effects on tumor growth were determined by tumor volume and histology analysis after 28 d administration of COE. The relative proportions of mature DCs and CD8+ T cells were measured in mononuclear cells that had been isolated from spleen by FACS. Results COE stimulated IL-2 and IFN-γ secretion of DCs, simultaneously enhanced the maturation of DCs by enhancing immunological molecule (CD40, CD80, CD86, H-2Kb, and I-Ab) expression in a dose-dependent manner. Furthermore, the chemotactic responses of DCs were significantly higher in COE-treated than untreated DCs, in association with higher chemokine receptor 7 expression. Furthermore, COE increased DCs produce IFN-γ and IL-2 in a dose-dependent manner when the concentration of COE less than 40 μg/mL, decreased DCs produce IL-10 and IL-4 also in a dose-dependent manner. In in vivo studies, COE can not only suppress growth of malignant hepatocellular carcinomas but also stimulate maturation of DCs, associated with strongly enhanced CD8+ CTL responses. Conclusion These data provide new insight into the mechanism of action of COE and indicate that the stimulation of maturation and function of DCs by COE contributes to its immunoregulatory effects.展开更多
Objective:To investigate the inhibitory effect of Celastrus orbiculatus extracts(COE)on the proliferation of lymphoma cells and the immune regulation ability on inflammation and thrombophilia in vivo.Methods:The 38B9 ...Objective:To investigate the inhibitory effect of Celastrus orbiculatus extracts(COE)on the proliferation of lymphoma cells and the immune regulation ability on inflammation and thrombophilia in vivo.Methods:The 38B9 lymphoma cells were treated with COE(160μg/mL)and CTX(25μmol/L).The apoptosis rate and cell cycle of each group were detected by flow cytometry.The secretion of inflammatory factors,including interleukin(IL)-6,IL-10,and tumor necrosis factorα(TNF-α),in cell supernatant was detected by enzyme-linked immunosorbent assay(ELISA).In vivo,BALB/c mice were subcutaneously injected with 38B9 lymphoma cells to establish lymphoma model.COE(3 mg·kg^(-1)·d^(-1))and CTX(40 mg·kg^(-1)·d^(-1))were administered to the model mice,respectively.The expression of plasma inflammatory factors(IL-6,IL-10 and TNF-α)and thrombus indexes,including D-dimer(D-D),von Willebrand factor(vWF)and tissue factor(TF),were detected by ELISA before tumor bearing(1 d),after tumor formation(14 d)and after intervention(21 d).PicoGreen dsDNA was used to detect the level of serum neutrophil extracellular traps(NETs).Flow cytometry was used to detect the expression of platelet activation marker calcium-dependent lectin-like receptor 2(CLEC-2).The tumor growth and survival of mice were recorded.Results:The 38B9 lymphoma cells were apoptotic after the intervention of COE and CTX.The ratio of G_(2)-M phase cells decreased in COE intervented cells compared with the control cells(P<0.05),and S phase cells decreased in CTX intervented cells(P<0.05).Also,the secretion level of IL-6 was significantly reduced after COE or CTX intervention(P<0.05),and IL-10 was significantly increased(P<0.05).Furthermore,the tumor mass was reduced,and the median survival time was longer in COE and CTX intervented tumor-bearing mice than in non-intervented mice.The significantly lower levels of TNF-α,IL-6,NETs,TF,DD and CLEC-2,as well as higher IL-10 were observed in COE and CTX treatment mice in comparision with the control mice(P<0.05).Conclusion:COE has a mil展开更多
Objective： The apoptosis of Hela cells induced by ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb was studied in order to assess its antitumor effect. Methods： Hela cells were cultured in ...Objective： The apoptosis of Hela cells induced by ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb was studied in order to assess its antitumor effect. Methods： Hela cells were cultured in vitro and treated by a series of concentrations of ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb. Cell proliferation was detected based on MTT assay. Quantity of apoptosis were observed and analyzed by flow cytometry with Annexin V and propidium iodide double staining. P53 gene expression was detected by flow cytometry. Results： The proliferation of Hela cells was obviously inhibited by 15, 30, 60 and 120 IJg/mL extract of Celastrus orbiculatus Thunb and apoptosis of Hela was induced by dosed dependent manner. P53 gene showed increasing tendency when treated by 60-480 IJg/mL extract of Celas- trus orbiculatus Thunb. Conclusion： The ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb could induce apoptosis of Hela gastric cancer cells by dose dependent manner, which maybe one of the important mechanisms of Celastrus orbiculatus Thunb＇s anticancer effects. P53 protein expression in Hela was up-regulated by Celastrus orbiculatus Thunb, which maybe one of the molecular mechanisms involved in the anticancer and proapoptotic effect of Celastrus or- biculatus Thunb.展开更多
[Objectives]This study was conducted to investigate the extraction technology of celastrol from Celastrus orbiculatus Thunb.[Methods]Solvent ultrasonic extraction was selected,and with the content of celastrol as the ...[Objectives]This study was conducted to investigate the extraction technology of celastrol from Celastrus orbiculatus Thunb.[Methods]Solvent ultrasonic extraction was selected,and with the content of celastrol as the evaluation index,the effects of different solvents,extraction time,temperatures and material-to-liquid ratios on the extraction rate of celastrol were investigated by single factor and orthogonal experiments.[Results]The optimal extraction conditions were as follows:a solvent ratio of petroleum ether to ethyl acetate at 1∶1,a ratio of solvent to material at 10∶1(v/w),extraction time of 30 min,and an extraction temperature at 30℃.[Conclusions]This method has high extraction rate,and is simple and feasible.展开更多
基金Supported by Plans of Colleges and Universities in Jiangsu Province to Postgraduate Research and Innovation(No.CX09B-321Z)State Administration of Traditional Chinese Medicine of People's Republic of China(No.04-05ZP35)
文摘Objective： Ce/astrus orbicu/atus Thunb. has been used for thousands of years in China as a remedy against cancer and inflammatory diseases. This study aims to investigate whether C. orbiculatus extract （COE） could inhibit angiogenesis, which is the pivotal step in tumor growth, invasiveness, and metastasis. Methods： In this study, the extract from the stem of C. orbiculatus was used. Mouse hepatic carcinoma cells （Hepat-6） were treated with COE in different nontoxic concentrations （10, 20, 40, 80, and 160 μg/mL）. The mRNA and protein expression levels of vascular endothelial growth factor （VEGF） were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot, respectively; the active fractions were further tested on C57BL/6 mice and human umbilical vein endothelial cells （HUVEC） for any antiangiogenic effects. Results： COE significantly inhibited proliferation and induced apoptosis in Hepal-6 cells and inhibited VEGF expression at both mRNA and protein levels. Furthermore, this agent inhibited the formation of the capillary-like structure in primary cultured HUVEC in a dose-dependent manner. In vivo, COE significantly reduced the volume and weight of solid tumors with low adverse effects and decreased tumor angiogenesis. Conclusions： In summary, COE could be used to treat hepatic carcinoma. The mechanisms of the antitumor activity of COE may be due to its effects against tumor angiogenesis by targeting the VEGF protein.
基金Supported by the National Natural Science Foundation of China(No.81403232 and No.81573656)Natural Science Foundation of Jiangsu Province(No.BK20171290 and No.BK2012686)Doctoral Fund of Ministry of Education of China(No.20133250120003)
文摘Objective: To evaluate the effects of Celastrus Orbiculatus extracts(COE) on metastasis in hypoxiainduced hepatocellular carcinoma cells(Hep G2) and to explore the underlying molecular mechanisms. Methods:The effect of COE(160, 200 and 240 μg/mL) on cell viability, scratch-wound, invasion and migration were studied by 3-4,5-dimethyl-2-thiazolyl-2,5-diphenyl-2-H-tetrazolium bromide(MTT), scratch-wound and transwell assays, respectively. Co Cl2 was used to establish a hypoxia model in vitro. Effects of COE on the expressions of E-cadherin, vimentin and N-cadherin were investigated with Western blot and immuno?uorescence analysis,respectively. Results: COE inhibited proliferation and metastasis of hypoxia-induced hepatocellular carcinoma cells in a dose-dependent manner(P<0.01). Furthermore, the expression of epithelial-mesenchymal transition(EMT) related markers were also remarkably suppressed in a dose-dependent manner(P<0.01). In addition, the upstream signaling pathways, including the hypoxia-inducible factor 1α(Hif-1α) and Twist1 were suppressed by COE. Additionally, the Hif-1α inhibitor 3-5'-hydroxymethyl-2'-furyl)-1-benzylindazole(YC-1), potently suppressed cell invasion and migration as well as expression of EMT in hypoxia-induced Hep G2 cells. Similarly, the combined treatment with COE and YC-1 showed a synergistic effect(P<0.01) compared with the treatment with COE or YC-1 alone in hypoxia-induced Hep G2 cells. Conclusions: COE signi?cantly inhibited the tumor metastasis and EMT by suppressing Hif-1α/Twist1 signaling pathway in hypoxia-induced Hep G2 cell. Thus, COE might have potential effect to inhibit the progression of Hep G2 in the context of tumor hypoxia.
基金supported by Plans of Colleges and Universities in Jiangsu Province to Postgraduate Research and Innovation (No.CX09B-321Z)State Administration of Traditional Chinese Medicine of People’s Republic of China (No. 04-05ZP35)
文摘Objective To discuss the antitumor mechanism preliminarily by observing effects of Celastrus orbiculatus extracts(COE) on vascular endothelial growth factor(VEGF) mRNA and protein expression in hepatoma(Hepa1-6) cells of mice.Methods Hepa1-6 cells were treated with COE at different nontoxic concentration(0,10,20,40,80,and 160 μg/mL) for 16 h.The mRNA and protein expressions of VEGF were detected by reverse transcription-PCR and Western Blotting,respectively.Results COE significantly inhibited VEGF expression at both mRNA and protein levels in a dose-dependent manner.Conclusion COE can inhibit VEGF expression in Hepa1-6 cells,therefore suggest that VEGF could be chosen as an therapeutic target for COE in the context of cancer chemoprevention and anticancer therapy.
基金Supported by the National Natural Science Foundation of China(No.81403232)the National Natural Science Foundation of Jiangsu Province(No.BK20171290,BK2012686)the Doctoral Fund of the Ministry of Education of China(No.20133250120003)
文摘Objective:To characterize the molecular mechanism underlying the antineoplastic activity of Celastrus orbiculatus Thunb.extracts(COE).Methods:The human hepatocellular carcinoma HepG2 cells with mammalian target of rapamycin(mTOR)knockdown expressed(HepG2/mTOR-)were constructed using molecular biological technology.In vitro,the HepG2/mTOR-cells were treated with COE at various concentrations(10,20,40,80,160 and 320μg/mL).Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assays.According to the half-maximal inhibitory concentration(IC50)value(140 mg/L),the concentrations of COE in the subsequent experiment was set to alleviate cytotoxicity.The HepG2/mTOR-cells were divided into 5 groups:negative control(untreated),COE treatment groups(40,80,120 mg/L COE)and positive control group(cisplatin,DDP,2 mg/L),respectively.Wild-type HepG2 cells were used as a blank control.The effects of COE on the cell apoptosis were analyzed by flow cytometry and transmission electronic microscopy(TEM),respectively.The protein expression levels of mTOR signal pathways were determined by Western blotting.In vivo,HepG2/mTOR-cells(2×106 cell/mice)were subcutaneously injected into the right flank of nude mice.Thirty-six female nude mice were randomly assigned to 6 groups according to body weight(6 mice per group)as follows:solvent vehicle control,Banmao Capsule treated group(BM,195 mg/kg),Tegafur,Gimeracil and Oteracil Potassium Capsules(10 mg/kg)treated group,and different dosages of COE(10,20,40 mg/kg)groups.Tumor growth was monitored and immunohistochemical staining was used to examine the expression of apoptosis-related proteins in tumor tissues.Results:COE inhibited the proliferation significantly in a concentration-dependent manner in HepG2/mTOR-cells(P<0.01).COE significantly induced the apoptosis of HepG2/mTOR-cells(P<0.01),and the apoptotic bodies can be observed under TEM.COE significantly inhibits the proteins expression of mTORrelated signal pathways.In vivo,COE significantly in
基金Plans of Colleges and Universities in Jiangsu Province to Postgraduate Research and Innovation (CX09B-321Z)State Administration of Traditional Chinese Medicine of P.R.China (04-05ZP35)
文摘Objective To examine the immunoregulation of Celastrus orbiculatus extracts (COE), a traditional Chinese medicine, on maturation and function of dendritic cells (DCs) in vitro and in vivo. Methods In vitro, after treated with COE in different nontoxic concentrations (0, 10, 20, 40, 80, and 160 μg/mL) for 5 d, the surface immunological molecules and cytokine secretion of mice bone marrow-derived DCs in response to COE were analyzed by flow cytometric analysis (FACS) and enzyme linked immunosorbent assay (ELISA), respectively. In vivo, mouse hepatoma cells (Hepa1-6, 1 × 106) were injected sc and were treated with different dosages of COE (10, 20 or 40 mg/kg/d). Effects on tumor growth were determined by tumor volume and histology analysis after 28 d administration of COE. The relative proportions of mature DCs and CD8+ T cells were measured in mononuclear cells that had been isolated from spleen by FACS. Results COE stimulated IL-2 and IFN-γ secretion of DCs, simultaneously enhanced the maturation of DCs by enhancing immunological molecule (CD40, CD80, CD86, H-2Kb, and I-Ab) expression in a dose-dependent manner. Furthermore, the chemotactic responses of DCs were significantly higher in COE-treated than untreated DCs, in association with higher chemokine receptor 7 expression. Furthermore, COE increased DCs produce IFN-γ and IL-2 in a dose-dependent manner when the concentration of COE less than 40 μg/mL, decreased DCs produce IL-10 and IL-4 also in a dose-dependent manner. In in vivo studies, COE can not only suppress growth of malignant hepatocellular carcinomas but also stimulate maturation of DCs, associated with strongly enhanced CD8+ CTL responses. Conclusion These data provide new insight into the mechanism of action of COE and indicate that the stimulation of maturation and function of DCs by COE contributes to its immunoregulatory effects.
基金Supported by Chinese and Western Medicine Cooperation Project of Subei People's Hospital(No.ZXXTGG2022BO3)Jiangsu Graduate Scientific Research Innovation Program(No.SJCX22 1813)。
文摘Objective:To investigate the inhibitory effect of Celastrus orbiculatus extracts(COE)on the proliferation of lymphoma cells and the immune regulation ability on inflammation and thrombophilia in vivo.Methods:The 38B9 lymphoma cells were treated with COE(160μg/mL)and CTX(25μmol/L).The apoptosis rate and cell cycle of each group were detected by flow cytometry.The secretion of inflammatory factors,including interleukin(IL)-6,IL-10,and tumor necrosis factorα(TNF-α),in cell supernatant was detected by enzyme-linked immunosorbent assay(ELISA).In vivo,BALB/c mice were subcutaneously injected with 38B9 lymphoma cells to establish lymphoma model.COE(3 mg·kg^(-1)·d^(-1))and CTX(40 mg·kg^(-1)·d^(-1))were administered to the model mice,respectively.The expression of plasma inflammatory factors(IL-6,IL-10 and TNF-α)and thrombus indexes,including D-dimer(D-D),von Willebrand factor(vWF)and tissue factor(TF),were detected by ELISA before tumor bearing(1 d),after tumor formation(14 d)and after intervention(21 d).PicoGreen dsDNA was used to detect the level of serum neutrophil extracellular traps(NETs).Flow cytometry was used to detect the expression of platelet activation marker calcium-dependent lectin-like receptor 2(CLEC-2).The tumor growth and survival of mice were recorded.Results:The 38B9 lymphoma cells were apoptotic after the intervention of COE and CTX.The ratio of G_(2)-M phase cells decreased in COE intervented cells compared with the control cells(P<0.05),and S phase cells decreased in CTX intervented cells(P<0.05).Also,the secretion level of IL-6 was significantly reduced after COE or CTX intervention(P<0.05),and IL-10 was significantly increased(P<0.05).Furthermore,the tumor mass was reduced,and the median survival time was longer in COE and CTX intervented tumor-bearing mice than in non-intervented mice.The significantly lower levels of TNF-α,IL-6,NETs,TF,DD and CLEC-2,as well as higher IL-10 were observed in COE and CTX treatment mice in comparision with the control mice(P<0.05).Conclusion:COE has a mil
基金Supported by a grant of State Administration of Traditional Special General Topics of Science and Technology (No. 04-05ZP35)
文摘Objective： The apoptosis of Hela cells induced by ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb was studied in order to assess its antitumor effect. Methods： Hela cells were cultured in vitro and treated by a series of concentrations of ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb. Cell proliferation was detected based on MTT assay. Quantity of apoptosis were observed and analyzed by flow cytometry with Annexin V and propidium iodide double staining. P53 gene expression was detected by flow cytometry. Results： The proliferation of Hela cells was obviously inhibited by 15, 30, 60 and 120 IJg/mL extract of Celastrus orbiculatus Thunb and apoptosis of Hela was induced by dosed dependent manner. P53 gene showed increasing tendency when treated by 60-480 IJg/mL extract of Celas- trus orbiculatus Thunb. Conclusion： The ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb could induce apoptosis of Hela gastric cancer cells by dose dependent manner, which maybe one of the important mechanisms of Celastrus orbiculatus Thunb＇s anticancer effects. P53 protein expression in Hela was up-regulated by Celastrus orbiculatus Thunb, which maybe one of the molecular mechanisms involved in the anticancer and proapoptotic effect of Celastrus or- biculatus Thunb.
文摘[Objectives]This study was conducted to investigate the extraction technology of celastrol from Celastrus orbiculatus Thunb.[Methods]Solvent ultrasonic extraction was selected,and with the content of celastrol as the evaluation index,the effects of different solvents,extraction time,temperatures and material-to-liquid ratios on the extraction rate of celastrol were investigated by single factor and orthogonal experiments.[Results]The optimal extraction conditions were as follows:a solvent ratio of petroleum ether to ethyl acetate at 1∶1,a ratio of solvent to material at 10∶1(v/w),extraction time of 30 min,and an extraction temperature at 30℃.[Conclusions]This method has high extraction rate,and is simple and feasible.
