Non-alcoholic steatohepatitis(NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK ...Non-alcoholic steatohepatitis(NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both in vivo and in vitro. The in vivo study was performed using male C57 BL/6 mice fed with methionine– choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The in vitro study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic acid plus palmitic acid, and adenovirus-down Cflar for 24 h,then treated with silibinin for 24 h. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibininsignificantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of Pparα, Fabp5, Cpt1α, Acox, Scd-1, Gpat and Mttp, reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2 E1 and CYP4 A in vivo.These effects were confirmed by the in vitro experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the β-oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity(CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity(CYP2 E1 and CYP4 A) to relieve oxidative stress.展开更多
Objective:The present work was to investigate the protective effects of the aqueous extract of Gynura procumbens(GPAE)against nonalcoholic steatohepatitis(NASH)in mice and NCTC-1469 cells.Methods:C57 BL/6 J mice were ...Objective:The present work was to investigate the protective effects of the aqueous extract of Gynura procumbens(GPAE)against nonalcoholic steatohepatitis(NASH)in mice and NCTC-1469 cells.Methods:C57 BL/6 J mice were fed with methionine and choline-deficient(MCD)diet and administered simultaneously with GPAE(500 and 1000 mg/kg/d,respectively)by gavage for six weeks.The biomarkers of NASH in serum and liver were determined.NCTC-1469 cells were pretreated with 0.25 mmol/L palmitic acid(PA)plus 0.5 mmol/L oleic acid(OA)for 24 h or treated with adenovirus expressing short-hairpin RNA against CFLAR(Ad-sh CFLAR)for 24 h and then treated with GPAE(80 and 160μg/m L,respectively)for 24 h,and the content of cellular biomarkers of NASH was detected.Results:In mice treated with MCD,GPAE could decrease the levels of serum ALT,AST,the content of hepatic TG,TC and MDA,repress the activities and protein expression of CYP2 E1 and CYP4 A and the phosphorylation of JNK,increase the activities of HO-1,CAT and GSH-Px,up-regulate the m RNA expression of PPARα,FABP5,CPT1α,ACOX,SCD-1,mGPAT,MTTP and the protein expression of CFLAR and NRF2.In NCTC-1469 cells treated with PA and OA,GPAE could decrease the content of cellular TG and ROS,promote the uptake of 2-NBDG,up-regulate the protein expression of CFLAR and NRF2.In NCTC-1469 cells treated with Ad-sh CFLAR,GPAE up-regulated the mRNA and protein expression of CFLAR,down-regulated the phosphorylation of JNK,and increased the protein expression of NRF2 and p IRS1.Conclusion:These results indicated that the activation on CFLAR-JNK pathway might be the main antiNASH mechanism of GPAE,which on the one hand promote theβ-oxidation and efflux of fatty acids in liver,and finally reduce hepatic lipid accumulation,on the other hand increase the activities of antioxidant enzymes and inhibit the activities of ROS generation enzymes by activating NRF2,and therefore attenuates hepatic oxidative stress damage.展开更多
非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)逐渐成为发病率最高的慢性肝病类型。NASH发病常常伴随全身代谢综合征,疾病进展具有发生肝硬化甚至肝癌的高风险。然而,目前临床上尚无一种获批的针对NASH的有效治疗药物。我们...非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)逐渐成为发病率最高的慢性肝病类型。NASH发病常常伴随全身代谢综合征,疾病进展具有发生肝硬化甚至肝癌的高风险。然而,目前临床上尚无一种获批的针对NASH的有效治疗药物。我们的最新研究结果发现,天然免疫重要分子CFLAR[CASP8 and FADD(Fas-associating protein with death domain)-like apoptosis regulator]直在NASH疾病进程中的关键负调控作用。深入的分子机制探索证实,CFLAR接靶向激酶MAP3K5[mitogen-activated protein kinase kinase kinase 5,也称为ASK1(apoptosis signal-regulating kinase 1)]并阻断其N-端二聚化,从而抑制ASK1和激酶MAPK8[mitogen-activated protein kinase 8,也称为JNK1(c-Jun N-terminal kinase 1)]的信号通路。此外,我们鉴定出源于CFLAR的一个小肽片段(S1)可以有效发挥CFLAR对ASK1的抑制作用。应用CFLAR(S1)治疗可有效改善并逆转小鼠和猴子中的NASH及并发的代谢综合征。综上所述,我们发现,CFLAR是控制NASH疾病进展的关键抑制子。CFLAR(S1)特异性抑制ASK 1激活的作用机制,为开发或筛选NASH的靶向治疗药物提供了可行的新方案。展开更多
基金supported by Major Technological Innovation Project of Hubei Province(Grant no.2016ACA140,China)Innovation and Entrepreneurship Training Project for College Students of the Ministry of Education(Grant no.201610512001,China)
文摘Non-alcoholic steatohepatitis(NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both in vivo and in vitro. The in vivo study was performed using male C57 BL/6 mice fed with methionine– choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The in vitro study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic acid plus palmitic acid, and adenovirus-down Cflar for 24 h,then treated with silibinin for 24 h. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibininsignificantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of Pparα, Fabp5, Cpt1α, Acox, Scd-1, Gpat and Mttp, reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2 E1 and CYP4 A in vivo.These effects were confirmed by the in vitro experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the β-oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity(CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity(CYP2 E1 and CYP4 A) to relieve oxidative stress.
基金supported by the major technological innovation project of Hubei Province(grant No.2016ACA140)the united fund for innovation and entrepreneurship of Ministry of Education of China(grant No.201610512001).
文摘Objective:The present work was to investigate the protective effects of the aqueous extract of Gynura procumbens(GPAE)against nonalcoholic steatohepatitis(NASH)in mice and NCTC-1469 cells.Methods:C57 BL/6 J mice were fed with methionine and choline-deficient(MCD)diet and administered simultaneously with GPAE(500 and 1000 mg/kg/d,respectively)by gavage for six weeks.The biomarkers of NASH in serum and liver were determined.NCTC-1469 cells were pretreated with 0.25 mmol/L palmitic acid(PA)plus 0.5 mmol/L oleic acid(OA)for 24 h or treated with adenovirus expressing short-hairpin RNA against CFLAR(Ad-sh CFLAR)for 24 h and then treated with GPAE(80 and 160μg/m L,respectively)for 24 h,and the content of cellular biomarkers of NASH was detected.Results:In mice treated with MCD,GPAE could decrease the levels of serum ALT,AST,the content of hepatic TG,TC and MDA,repress the activities and protein expression of CYP2 E1 and CYP4 A and the phosphorylation of JNK,increase the activities of HO-1,CAT and GSH-Px,up-regulate the m RNA expression of PPARα,FABP5,CPT1α,ACOX,SCD-1,mGPAT,MTTP and the protein expression of CFLAR and NRF2.In NCTC-1469 cells treated with PA and OA,GPAE could decrease the content of cellular TG and ROS,promote the uptake of 2-NBDG,up-regulate the protein expression of CFLAR and NRF2.In NCTC-1469 cells treated with Ad-sh CFLAR,GPAE up-regulated the mRNA and protein expression of CFLAR,down-regulated the phosphorylation of JNK,and increased the protein expression of NRF2 and p IRS1.Conclusion:These results indicated that the activation on CFLAR-JNK pathway might be the main antiNASH mechanism of GPAE,which on the one hand promote theβ-oxidation and efflux of fatty acids in liver,and finally reduce hepatic lipid accumulation,on the other hand increase the activities of antioxidant enzymes and inhibit the activities of ROS generation enzymes by activating NRF2,and therefore attenuates hepatic oxidative stress damage.
文摘非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)逐渐成为发病率最高的慢性肝病类型。NASH发病常常伴随全身代谢综合征,疾病进展具有发生肝硬化甚至肝癌的高风险。然而,目前临床上尚无一种获批的针对NASH的有效治疗药物。我们的最新研究结果发现,天然免疫重要分子CFLAR[CASP8 and FADD(Fas-associating protein with death domain)-like apoptosis regulator]直在NASH疾病进程中的关键负调控作用。深入的分子机制探索证实,CFLAR接靶向激酶MAP3K5[mitogen-activated protein kinase kinase kinase 5,也称为ASK1(apoptosis signal-regulating kinase 1)]并阻断其N-端二聚化,从而抑制ASK1和激酶MAPK8[mitogen-activated protein kinase 8,也称为JNK1(c-Jun N-terminal kinase 1)]的信号通路。此外,我们鉴定出源于CFLAR的一个小肽片段(S1)可以有效发挥CFLAR对ASK1的抑制作用。应用CFLAR(S1)治疗可有效改善并逆转小鼠和猴子中的NASH及并发的代谢综合征。综上所述,我们发现,CFLAR是控制NASH疾病进展的关键抑制子。CFLAR(S1)特异性抑制ASK 1激活的作用机制,为开发或筛选NASH的靶向治疗药物提供了可行的新方案。
