AIM: To study the growth inhibitory and apoptotic effects of Scutellaria barbata D.Don (S. barbata) and to determine the underlying mechanism of its antiturnor activity in mouse liver cancer cell line H22.METHODS:...AIM: To study the growth inhibitory and apoptotic effects of Scutellaria barbata D.Don (S. barbata) and to determine the underlying mechanism of its antiturnor activity in mouse liver cancer cell line H22.METHODS: Proliferation of H22 cells was examined by MTT assay. Cellular morphology of PC-2 cells was observed under fluorescence microscope and transmission electron microscope (EM). Mitochondrial transmembrane potential was determined under laser scanning confocal microscope (LSCM) with rhodamine 123 staining. Flow cytometry was performed to analyze the cell cycle of H22 cells with propidium iodide staining. Protein level of cytochrome C and caspase-3 was measured by semi-quantitive RT-PCR and Western blot analysis. Activity of caspase-3 enzyme was measured by spectrofluorometrv.RESULTS: M-I-I- assay showed that extracts from S. barbata (ESB) could inhibit the proliferation of H22 cells in a time-dependent manner. Among the various phasesof cell cycle, the percentage of cells in S phase was significantly decreased, while the percentage of cells in G1 phase was increased. Flow cytometry assay also showed that ESB had a positive effect on apoptosis. Typical apoptotic morphologies such as condensation and fragmentation of nuclei and blebbing membrane of apoptotic cells could be observed under transmission electron microscope and fluorescence microscope. To further investige the molecular mechanism behind ESB-induced apoptosis, ESB-treated cells rapidly lost their mitochondrial transmembrane potential, released mitochondrial cytochrome C into cytosol, and induced caspase-3 activity in a dose-dependent manner. CONCLUSION: ESB can effectively inhibit the proliferation and induce apoptosis of H22 cells involving loss of mitochondrial transmembrane potential, release of cytochrome C, and activation of caspase-3.展开更多
A practical solution of qualitatively analyzing quaternary alkaloids in coptis-scute herb couple by electrospray ionization mass spectrometry(ES1-MS) was developed. Without the complicated pretreatment of sample, th...A practical solution of qualitatively analyzing quaternary alkaloids in coptis-scute herb couple by electrospray ionization mass spectrometry(ES1-MS) was developed. Without the complicated pretreatment of sample, thc active ingredients including berberine, palmatine, coptisine, jatrorrhizine, epiberberine, and columbamine were identified and some relative content changing rules of alkaloids in coptis-scute couple were summarized in this article. The overall profiles of the complex extracts were obtained. After adding an internal standard(rutaecarpine), semi-quantitative analysis was performed and the result indicates that the actual content of alkaloids was decreased by increasing the amount of scute. Based on the data obtained by high-performance capillary electrophoresis(HPCE), the feasibility of semi-quantitative analysis by ES1-MS was further proved.展开更多
Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the ...Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the peripheral circulation of the rats before and after the treatment.Methods: Setaria cer vi worms were implanted in white rats via laparotomy and microfilaraemic rats were divided into 4 groups. Groups 1, 2 and 3 were treated with DEC, tetramisole and chlorpromazine respectively, while Group 4 served as infected control. Longevity of microi lariae and dif erential leucocyte counts were recorded till the disappearance of microi lariae from peripheral blood. Glucose-6-phosphatase and succinate dehydrogenase enzymes were localized in the microi lariae recovered from normal and treated rats.Results: The microi lariae survived for 48 days in untreated rats while survival was reduced to 15, 21 and 27 days after treatment with DEC, tetramisole and chlorpromazine, respectively. Eosinophils and neutrophils increased during 2nd and 3rd weeks, whereas the lymphocytes increased during 4-7 weeks. DEC treatment resulted in slight decrease in the localization of succinate dehydrogenase but not in glucose-6-phosphatase. Tetramisole and chlorpromazine treatment did not show any appreciable change in the localization of both the above enzymes. Conclusions: DEC proved the most ef ective drug which cleared the microi laraemia within 15 days and reduced the activity of succinate dehydrogenase to some extent followed by tetramisole and chlorpromazine which took more time for the clearance of microi lariae and had no ef ect on the localization of both glucose-6-phosphatase and succinate dehydrogenase.展开更多
基金Supported by The Science and Technology Foundation of Shaanxi Province, China, No. 2006K16-G5(1) Sci-tech Program of Xi’an City, China, No. YF07175
文摘AIM: To study the growth inhibitory and apoptotic effects of Scutellaria barbata D.Don (S. barbata) and to determine the underlying mechanism of its antiturnor activity in mouse liver cancer cell line H22.METHODS: Proliferation of H22 cells was examined by MTT assay. Cellular morphology of PC-2 cells was observed under fluorescence microscope and transmission electron microscope (EM). Mitochondrial transmembrane potential was determined under laser scanning confocal microscope (LSCM) with rhodamine 123 staining. Flow cytometry was performed to analyze the cell cycle of H22 cells with propidium iodide staining. Protein level of cytochrome C and caspase-3 was measured by semi-quantitive RT-PCR and Western blot analysis. Activity of caspase-3 enzyme was measured by spectrofluorometrv.RESULTS: M-I-I- assay showed that extracts from S. barbata (ESB) could inhibit the proliferation of H22 cells in a time-dependent manner. Among the various phasesof cell cycle, the percentage of cells in S phase was significantly decreased, while the percentage of cells in G1 phase was increased. Flow cytometry assay also showed that ESB had a positive effect on apoptosis. Typical apoptotic morphologies such as condensation and fragmentation of nuclei and blebbing membrane of apoptotic cells could be observed under transmission electron microscope and fluorescence microscope. To further investige the molecular mechanism behind ESB-induced apoptosis, ESB-treated cells rapidly lost their mitochondrial transmembrane potential, released mitochondrial cytochrome C into cytosol, and induced caspase-3 activity in a dose-dependent manner. CONCLUSION: ESB can effectively inhibit the proliferation and induce apoptosis of H22 cells involving loss of mitochondrial transmembrane potential, release of cytochrome C, and activation of caspase-3.
基金Supported by the National Basic Research Program of China(No.2006CB504706)the National Natural Science Foundation of China(No.30672600)+1 种基金the Great Research Project of Chinese Academy of Sciences(No.KGCX2-SW-213-06)the Key Project of Jilin Provinical Science and Technology, China(No.20060902)
文摘A practical solution of qualitatively analyzing quaternary alkaloids in coptis-scute herb couple by electrospray ionization mass spectrometry(ES1-MS) was developed. Without the complicated pretreatment of sample, thc active ingredients including berberine, palmatine, coptisine, jatrorrhizine, epiberberine, and columbamine were identified and some relative content changing rules of alkaloids in coptis-scute couple were summarized in this article. The overall profiles of the complex extracts were obtained. After adding an internal standard(rutaecarpine), semi-quantitative analysis was performed and the result indicates that the actual content of alkaloids was decreased by increasing the amount of scute. Based on the data obtained by high-performance capillary electrophoresis(HPCE), the feasibility of semi-quantitative analysis by ES1-MS was further proved.
基金Supported by the Deanship of Scientific Research(DSR),King Abdulaziz University,Jeddah(Grant No.407/142/1434)
文摘Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the peripheral circulation of the rats before and after the treatment.Methods: Setaria cer vi worms were implanted in white rats via laparotomy and microfilaraemic rats were divided into 4 groups. Groups 1, 2 and 3 were treated with DEC, tetramisole and chlorpromazine respectively, while Group 4 served as infected control. Longevity of microi lariae and dif erential leucocyte counts were recorded till the disappearance of microi lariae from peripheral blood. Glucose-6-phosphatase and succinate dehydrogenase enzymes were localized in the microi lariae recovered from normal and treated rats.Results: The microi lariae survived for 48 days in untreated rats while survival was reduced to 15, 21 and 27 days after treatment with DEC, tetramisole and chlorpromazine, respectively. Eosinophils and neutrophils increased during 2nd and 3rd weeks, whereas the lymphocytes increased during 4-7 weeks. DEC treatment resulted in slight decrease in the localization of succinate dehydrogenase but not in glucose-6-phosphatase. Tetramisole and chlorpromazine treatment did not show any appreciable change in the localization of both the above enzymes. Conclusions: DEC proved the most ef ective drug which cleared the microi laraemia within 15 days and reduced the activity of succinate dehydrogenase to some extent followed by tetramisole and chlorpromazine which took more time for the clearance of microi lariae and had no ef ect on the localization of both glucose-6-phosphatase and succinate dehydrogenase.