OBJECTIVE:To explore the mechanism of the Chinese medicine Cigu Xiaozhi prescription(慈菇消脂方,CGXZ)in the treatment of the non-alcoholic fatty liver disease(NAFLD)by detoxification and phlegm-reducing,the effect of ...OBJECTIVE:To explore the mechanism of the Chinese medicine Cigu Xiaozhi prescription(慈菇消脂方,CGXZ)in the treatment of the non-alcoholic fatty liver disease(NAFLD)by detoxification and phlegm-reducing,the effect of CGXZ prescription on ceramide-mediated lipid apoptosis in Hep G2 cells with NAFLD.METHODS:The experiment was randomly divided into 6groups:normal control group,model group,CGXZ prescription medicated serum high,medium,and low dose groups,and pioglitazone positive control group.Using 500μmol/L free fatty acid(FFA)mixture to induce Hep G2 cells to establish NAFLD cell model,respectively,with 2%,4%,and 6%concentration of CGXZ prescription medicated serum intervention for 24 h.The changes in organelles and lipid droplet accumulation were observed under the electron microscope.Furthermore,Td T-mediated d UTP Nick-End Labeling method was used to assay hepatocyte apoptosis;Biochemical determination of glutamic-pyruvic transaminase,glutamic oxalacetic transaminase,triglycerides,and FFA levels in Hep G2cells;the content of ceramide was determined by highperformance thin-layer chromatography.Finally,Western Blot and quantitative real-time polymerase chain reaction(q RT-PCR)were used to determine the protein and gene expression levels,such as inducible nitric oxide synthase(i NOS),nuclear factorκB(NF-κB),B cell lymphoma 2(Bcl-2)and Bcl-2-associated X(Bax).RESULTS:Under the electron microscope,the cells in the model group showed moderate-to-severe steatosis,and apoptotic bodies could be seen.The model group had greater improvements in the apoptosis rate(P<0.01),and the levels of ceramide C2 and FFA in the cytoplasm(P<0.01)than the normal control group.The protein expressions of NF-κB,i NOS,and Bax were significantly up-regulated(P<0.05),while the Bcl-2 had no significant change(P>0.05).Compared with the model group,the levels of ceramide C2 and FFA(P<0.01),the protein expressions of NF-κB,i NOS,and Bax(P<0.05)in the CGXZ prescription treatment group and pioglitazone positive control group were signific展开更多
OBJECTIVE:To explore the mechanistic effects of Yajieshaba(YJSB) on enhanced liver detoxification.METHODS:The effects of YJSB on alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were assayed in five a...OBJECTIVE:To explore the mechanistic effects of Yajieshaba(YJSB) on enhanced liver detoxification.METHODS:The effects of YJSB on alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were assayed in five acute chemical liver injury models[carbon tetrachloride(CCU),D-galactosamine(D-Glan),4-acetamidophenol(AAP),thioacetamide(TAA) and 1-naphthyl isothiocyanate(ANIT)].Sleep latency and sleep time of pentobarbital sodium were tested in control mice and CCL model miceafter oral YJSB administration.The effects of YJSB on drug metabolism enzymes of liver microsomes were tested in control rats and CCI_4model rats.The levels of cytochrome P450(CYP450) and Cyt b5 in liver microsomes were assayed using the method by Omura and Sato,and activities of erythromycin N-demethylase(ERD)and aminopyrine N-demethyl(ADM) were evaluated by Nash colorimetry.Probe substrate-based high performance liquid chromatography(HPLC)methods were established for CYP3A4 and CYP1A2.RESULTS:The level of serum ALT was reduced by YJSB at 3.51 g/kg in the five models as follows:CCl_4 > D-Glan,AAP,ANIT > TAA.YJSB treatment did not reduce the level of serum AST.YJSB at 3.51 g/kg prolonged the sleep latency in control mice and shortened the sleep time of control mice and CCl_4 model mice.For control rats,YJSB at 2.43 g/kg increased the levels of CYP450 and Cyt b5 and induced the activities of ERD and ADM;for liver injuries induced by CCI_4 in rats,YJSB at 2.43 g/kg increasedthe levels of CYP450 and Cyt b5.These results suggest that YJSB at 2.43 g/kg induces CYP3A4 and CYP1A2.CONCLUSION:These results suggest that YJSB enhanced liver detoxification and the mechanisms may be partially related to CYP3A4 and CYP1A2 induction.展开更多
基金Natural Science Foundation-funded Project:the Interaction of mi RNAs and Hh Signaling Pathway in NASH Related Liver Fibrosis and the Intervention of Sagittaria Xiaozhi Pill(No.81860821)Toxin and Eliminating Phlegm Intervention Ceramide and Induced iNOS Nonalcoholic Fatty Liver Disease Fat Research of Apoptosis Signaling(No.81460710)。
文摘OBJECTIVE:To explore the mechanism of the Chinese medicine Cigu Xiaozhi prescription(慈菇消脂方,CGXZ)in the treatment of the non-alcoholic fatty liver disease(NAFLD)by detoxification and phlegm-reducing,the effect of CGXZ prescription on ceramide-mediated lipid apoptosis in Hep G2 cells with NAFLD.METHODS:The experiment was randomly divided into 6groups:normal control group,model group,CGXZ prescription medicated serum high,medium,and low dose groups,and pioglitazone positive control group.Using 500μmol/L free fatty acid(FFA)mixture to induce Hep G2 cells to establish NAFLD cell model,respectively,with 2%,4%,and 6%concentration of CGXZ prescription medicated serum intervention for 24 h.The changes in organelles and lipid droplet accumulation were observed under the electron microscope.Furthermore,Td T-mediated d UTP Nick-End Labeling method was used to assay hepatocyte apoptosis;Biochemical determination of glutamic-pyruvic transaminase,glutamic oxalacetic transaminase,triglycerides,and FFA levels in Hep G2cells;the content of ceramide was determined by highperformance thin-layer chromatography.Finally,Western Blot and quantitative real-time polymerase chain reaction(q RT-PCR)were used to determine the protein and gene expression levels,such as inducible nitric oxide synthase(i NOS),nuclear factorκB(NF-κB),B cell lymphoma 2(Bcl-2)and Bcl-2-associated X(Bax).RESULTS:Under the electron microscope,the cells in the model group showed moderate-to-severe steatosis,and apoptotic bodies could be seen.The model group had greater improvements in the apoptosis rate(P<0.01),and the levels of ceramide C2 and FFA in the cytoplasm(P<0.01)than the normal control group.The protein expressions of NF-κB,i NOS,and Bax were significantly up-regulated(P<0.05),while the Bcl-2 had no significant change(P>0.05).Compared with the model group,the levels of ceramide C2 and FFA(P<0.01),the protein expressions of NF-κB,i NOS,and Bax(P<0.05)in the CGXZ prescription treatment group and pioglitazone positive control group were signific
基金Supported by the Natural Science Foundation of China(Detoxification Mechanisms of Dai Medicine Ya-Jie-Sha-Ba In Food Poisoning and Drug Toxicity,No.81160555)Key Project of Science Foundation by the Department of Education,Yunnan Province,China(Detoxification Mechanisms of Dai Medicine Ya-Jie-Sha-Ba,No.ZD201008)12th Five-year Key Construction Discipline of State Administration of Traditional Chinese Medicine"Dai Pharmacy"
文摘OBJECTIVE:To explore the mechanistic effects of Yajieshaba(YJSB) on enhanced liver detoxification.METHODS:The effects of YJSB on alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were assayed in five acute chemical liver injury models[carbon tetrachloride(CCU),D-galactosamine(D-Glan),4-acetamidophenol(AAP),thioacetamide(TAA) and 1-naphthyl isothiocyanate(ANIT)].Sleep latency and sleep time of pentobarbital sodium were tested in control mice and CCL model miceafter oral YJSB administration.The effects of YJSB on drug metabolism enzymes of liver microsomes were tested in control rats and CCI_4model rats.The levels of cytochrome P450(CYP450) and Cyt b5 in liver microsomes were assayed using the method by Omura and Sato,and activities of erythromycin N-demethylase(ERD)and aminopyrine N-demethyl(ADM) were evaluated by Nash colorimetry.Probe substrate-based high performance liquid chromatography(HPLC)methods were established for CYP3A4 and CYP1A2.RESULTS:The level of serum ALT was reduced by YJSB at 3.51 g/kg in the five models as follows:CCl_4 > D-Glan,AAP,ANIT > TAA.YJSB treatment did not reduce the level of serum AST.YJSB at 3.51 g/kg prolonged the sleep latency in control mice and shortened the sleep time of control mice and CCl_4 model mice.For control rats,YJSB at 2.43 g/kg increased the levels of CYP450 and Cyt b5 and induced the activities of ERD and ADM;for liver injuries induced by CCI_4 in rats,YJSB at 2.43 g/kg increasedthe levels of CYP450 and Cyt b5.These results suggest that YJSB at 2.43 g/kg induces CYP3A4 and CYP1A2.CONCLUSION:These results suggest that YJSB enhanced liver detoxification and the mechanisms may be partially related to CYP3A4 and CYP1A2 induction.
