[Objectives] To study the effects of Roudoukou-8 San extract on hydrogen peroxide( H_2O_2) induced cardiomyocyte injury of rats and explore its action mechanism. [Methods] The cardiomyocytes of neonatal rats were isol...[Objectives] To study the effects of Roudoukou-8 San extract on hydrogen peroxide( H_2O_2) induced cardiomyocyte injury of rats and explore its action mechanism. [Methods] The cardiomyocytes of neonatal rats were isolated and cultured,and the H_2O_2 induced cardiomyocyte injury model was established. Methyl thiazolyl tetrazolium( MTT) assay was used to detect the protective effects of Roudoukou-8 San extract on H_2O_2 induced cardiomyocyte. The effects of Roudoukou-8 San on cardiomyocyte morphology were observed under inverted microscope. The contents of lactate dehydrogenase( LDH),creatine kinase( CK) and aspartate aminotransferase( AST) in cell culture medium were determined by automatic biochemical instrument; the levels of malondialdehyde( MDA),superoxide dismutase( SOD) and nitric oxide( NO) in the cells were detected by kit method. The apoptotic morphology of cardiomyocytes was observed by Hoechst fluorescence staining.Cell apoptosis were measured by Annexin V and PI double staining and flow cytometer. [Results]100 μmol/L of H_2O_2 acting 2 h could cause about 50% of cardiomyocyte injury. H_2O_2 model group showed increased cell gap,decreased cell count,cell cytoplasmic vacuoles and other obvious damages. Compared with H_2O_2 model group,in Roudoukou-8 San extract dose groups,cardiomyocyte morphology showed different degrees of improvement,Roudoukou-8 San extract can significantly reduce the content of LDH,CK and AST in H_2O_2 injured myocardial cell culture solution,significantly reduce content of MDA and NO in H_2O_2 injured myocardial cells,increase the SOD activity,and significantly inhibit the apoptosis of H_2O_2 injured myocardial cells. [Conclusions]Through improving the cell survival status,increasing the cell viability,reducing oxidative stress injury,inhibiting inflammatory responses,and inhibiting apoptosis,Roudoukou-8 San extract can improve the state of H_2O_2 induced cardiomyocyte injury,so as to protect H_2O_2 injured myocardial cells.展开更多
To study the inhibitory effect of Huqi San （Qi- protecting powder） on rat prehepatocarcinoma induced by diethylinitrosamine （DEN） by analyzing the mutational activation of c-fos proto-oncogene and over-expression ...To study the inhibitory effect of Huqi San （Qi- protecting powder） on rat prehepatocarcinoma induced by diethylinitrosamine （DEN） by analyzing the mutational activation of c-fos proto-oncogene and over-expression of c-jun and c-myc oncogenes. METHODS： A Solt-Farber two-step test model of prehepatocarcinoma was induced in rats by DEN and 2-acetylaminofluorene （AAF） to investigate the modifying effects of Huqi San on the expression of c-jun, c-fos and c-myc in DEN-mediated hepatocarcinogenesis. Huqi San was made of eight medicinal herbs containing glycoprival granules, in which each milliliter contains 0.38 g crude drugs. T-glutamy-transpeptidase-isoenzyme （T-GTase） was determined with histochemical methods. Level of 8-hydroxydeoxyguanosine （OHdG） formed in liver and c-jun, c-fos and c-myc proto-oncogenes were detected by immunohistochemical methods. RESULTS： The level of 8-OHdG, a mark of oxidative DNA damage, was significantly decreased in the liver of rats with prehepatocarcinoma induced by DEN who received 8 g/kg body weight or 4 g/kg body weight Huqi San before （1 wk） and after DEN exposure （4 wk）. Huqi San- treated rats showed a significant decrease in number of T-GT positive foci （P 〈 0.001, prevention group： 4.96-0.72 vs 29.46-2.17; large dose therapeutic group： 7.53-0.88 vs 29.46-2.17）. On the other hand, significant changes in expression of c-jun, c-fos and c-myc were found in Huqi San-treated rats. CONCLUSION： Activation of c-jun, c-fos and c-myc plays a crucial role in the pathogenesis of liver cancer.Huqi San can inhibit the over-expression of c-jun, c-fos and c-myc oncogenes and liver preneolastic lesions induced by DEN.展开更多
基金Supported by Project of National Natural Science Foundation(81360678)Program for Young Talents of Science and Technology in Universities of Inner Mongolia(NJYT-17-B29)Program of Health and Family Planning Commission of Inner Mongolia(201702059)
文摘[Objectives] To study the effects of Roudoukou-8 San extract on hydrogen peroxide( H_2O_2) induced cardiomyocyte injury of rats and explore its action mechanism. [Methods] The cardiomyocytes of neonatal rats were isolated and cultured,and the H_2O_2 induced cardiomyocyte injury model was established. Methyl thiazolyl tetrazolium( MTT) assay was used to detect the protective effects of Roudoukou-8 San extract on H_2O_2 induced cardiomyocyte. The effects of Roudoukou-8 San on cardiomyocyte morphology were observed under inverted microscope. The contents of lactate dehydrogenase( LDH),creatine kinase( CK) and aspartate aminotransferase( AST) in cell culture medium were determined by automatic biochemical instrument; the levels of malondialdehyde( MDA),superoxide dismutase( SOD) and nitric oxide( NO) in the cells were detected by kit method. The apoptotic morphology of cardiomyocytes was observed by Hoechst fluorescence staining.Cell apoptosis were measured by Annexin V and PI double staining and flow cytometer. [Results]100 μmol/L of H_2O_2 acting 2 h could cause about 50% of cardiomyocyte injury. H_2O_2 model group showed increased cell gap,decreased cell count,cell cytoplasmic vacuoles and other obvious damages. Compared with H_2O_2 model group,in Roudoukou-8 San extract dose groups,cardiomyocyte morphology showed different degrees of improvement,Roudoukou-8 San extract can significantly reduce the content of LDH,CK and AST in H_2O_2 injured myocardial cell culture solution,significantly reduce content of MDA and NO in H_2O_2 injured myocardial cells,increase the SOD activity,and significantly inhibit the apoptosis of H_2O_2 injured myocardial cells. [Conclusions]Through improving the cell survival status,increasing the cell viability,reducing oxidative stress injury,inhibiting inflammatory responses,and inhibiting apoptosis,Roudoukou-8 San extract can improve the state of H_2O_2 induced cardiomyocyte injury,so as to protect H_2O_2 injured myocardial cells.
基金Supported by The Chinese Medicine Technology Item of Peking City (No.JJ2005-10)Beijing Municipal Commission of Education (No. M200610025003)
文摘To study the inhibitory effect of Huqi San （Qi- protecting powder） on rat prehepatocarcinoma induced by diethylinitrosamine （DEN） by analyzing the mutational activation of c-fos proto-oncogene and over-expression of c-jun and c-myc oncogenes. METHODS： A Solt-Farber two-step test model of prehepatocarcinoma was induced in rats by DEN and 2-acetylaminofluorene （AAF） to investigate the modifying effects of Huqi San on the expression of c-jun, c-fos and c-myc in DEN-mediated hepatocarcinogenesis. Huqi San was made of eight medicinal herbs containing glycoprival granules, in which each milliliter contains 0.38 g crude drugs. T-glutamy-transpeptidase-isoenzyme （T-GTase） was determined with histochemical methods. Level of 8-hydroxydeoxyguanosine （OHdG） formed in liver and c-jun, c-fos and c-myc proto-oncogenes were detected by immunohistochemical methods. RESULTS： The level of 8-OHdG, a mark of oxidative DNA damage, was significantly decreased in the liver of rats with prehepatocarcinoma induced by DEN who received 8 g/kg body weight or 4 g/kg body weight Huqi San before （1 wk） and after DEN exposure （4 wk）. Huqi San- treated rats showed a significant decrease in number of T-GT positive foci （P 〈 0.001, prevention group： 4.96-0.72 vs 29.46-2.17; large dose therapeutic group： 7.53-0.88 vs 29.46-2.17）. On the other hand, significant changes in expression of c-jun, c-fos and c-myc were found in Huqi San-treated rats. CONCLUSION： Activation of c-jun, c-fos and c-myc plays a crucial role in the pathogenesis of liver cancer.Huqi San can inhibit the over-expression of c-jun, c-fos and c-myc oncogenes and liver preneolastic lesions induced by DEN.
