目的:研究多黏菌素B通过下调核因子κB(nuclear factor-κB,NF-κB)对乙型肝炎病毒(hepatitis B virus,HBV)转录和复制的影响。方法:通过MTT法检测多黏菌素B在HepG2-NTCP、HepAD38、HepG2.2.15、HepG2、Huh-7和PLC/PRF/5细胞中的细胞毒...目的:研究多黏菌素B通过下调核因子κB(nuclear factor-κB,NF-κB)对乙型肝炎病毒(hepatitis B virus,HBV)转录和复制的影响。方法:通过MTT法检测多黏菌素B在HepG2-NTCP、HepAD38、HepG2.2.15、HepG2、Huh-7和PLC/PRF/5细胞中的细胞毒性;使用10、50和100 nmol/L的多黏菌素B处理HBV感染的HepG2-NTCP细胞和HBV稳定复制的HepG2.2.15细胞,通过RT-qPCR方法检测细胞中HBV RNA和HBV DNA水平,ELISA方法检测细胞上清中乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)和乙型肝炎e抗原(hepatitis B e antigen,HBeAg)水平,明确多黏菌素B对HBV的调控作用。利用转录组测序筛选多黏菌素B调控HBV转录复制的效应分子NF-κB;同时检测敲减及过表达NF-κB对HBV的调控作用;利用双萤光素酶报告基因实验检测NF-κB对Cp、Xp、Sp1和Sp2启动子活性的影响;利用回补实验探究多黏菌素B调控HBV转录复制是否依赖于NF-κB。结果:MTT实验结果显示,浓度低于100μmol/L时,多黏菌素B对HepG2-NTCP、HepAD38、HepG2.2.15、HepG2、Huh-7和PLC/PRF/5细胞无明显毒性。在10、50和100 nmol/L的浓度下,多黏菌素B呈浓度依赖性抑制HBV感染的HepG2-NTCP细胞和HBV稳定复制的HepG2.2.15细胞中HBV RNA、HBV DNA、HBsAg和HBeAg水平。通过转录组测序发现,多黏菌素B可以显著抑制HBV感染的HepG2-NTCP细胞中NF-κB的表达水平;进一步,过表达NF-κB显著上调HBV感染的HepG2-NTCP细胞中HBV RNA、HBV DNA、HBsAg和HBeAg水平,沉默时则相反;同时,双萤光素酶报告基因实验显示NF-κB可增强HBV Sp2启动子活性。回补实验发现多黏菌素B调控HBV转录复制依赖于NF-κB。结论:多黏菌素B通过下调NF-κB进而抑制HBV Sp2启动子活性,最终抑制HBV转录复制。展开更多
OBJECTIVE Liver fibrosis is a chronic damage process related to the further progression of hepatic cirrhosis and has yet no truly effective treatment is available.This study aimed to investigate the effects of isochlo...OBJECTIVE Liver fibrosis is a chronic damage process related to the further progression of hepatic cirrhosis and has yet no truly effective treatment is available.This study aimed to investigate the effects of isochlorogenic acid A(ICQA)on liver fibrosis induced by carbon tetrachloride(CCl4)and clarify the underlying mechanism.METHODS Rats were treated with CCl4 for eight weeks in order to induce liver fibrosis and simultaneously orally administered with ICQA(10,20 and 40 mg·kg-1).RESULTS ICQA had significant protective effect on liver injury,inflammation,and fibrosis in rats.Meanwhile,ICQA prevented the activation of hepatic stellate cells(HSC)as indicated by inhibiting the overexpres⁃sion of a-smooth muscle actin(a-SMA).In addition,reduced fibrosis was found to be associated with decreased protein expression of high-mobility group box 1(HMGB1)and toll like receptor(TLR)4.Moreover,ICQA supressed the cytoplasmic translocation of HMGB1 in rat liver.Further investigations indicated that ICQA treatment significantly attenuated nuclear translocation of the nuclear factor-κB(NF-κB)p65 and inhibited degradation of IkBa expression in the liver of rats with liver fibrosis.CONCLUSION ICQA has hepatoprotective and anti-fibrotic effects in rats with liver fibrosis through modulating the HMGB1/TLR4/NF-κB signaling pathways.展开更多
文摘目的:研究多黏菌素B通过下调核因子κB(nuclear factor-κB,NF-κB)对乙型肝炎病毒(hepatitis B virus,HBV)转录和复制的影响。方法:通过MTT法检测多黏菌素B在HepG2-NTCP、HepAD38、HepG2.2.15、HepG2、Huh-7和PLC/PRF/5细胞中的细胞毒性;使用10、50和100 nmol/L的多黏菌素B处理HBV感染的HepG2-NTCP细胞和HBV稳定复制的HepG2.2.15细胞,通过RT-qPCR方法检测细胞中HBV RNA和HBV DNA水平,ELISA方法检测细胞上清中乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)和乙型肝炎e抗原(hepatitis B e antigen,HBeAg)水平,明确多黏菌素B对HBV的调控作用。利用转录组测序筛选多黏菌素B调控HBV转录复制的效应分子NF-κB;同时检测敲减及过表达NF-κB对HBV的调控作用;利用双萤光素酶报告基因实验检测NF-κB对Cp、Xp、Sp1和Sp2启动子活性的影响;利用回补实验探究多黏菌素B调控HBV转录复制是否依赖于NF-κB。结果:MTT实验结果显示,浓度低于100μmol/L时,多黏菌素B对HepG2-NTCP、HepAD38、HepG2.2.15、HepG2、Huh-7和PLC/PRF/5细胞无明显毒性。在10、50和100 nmol/L的浓度下,多黏菌素B呈浓度依赖性抑制HBV感染的HepG2-NTCP细胞和HBV稳定复制的HepG2.2.15细胞中HBV RNA、HBV DNA、HBsAg和HBeAg水平。通过转录组测序发现,多黏菌素B可以显著抑制HBV感染的HepG2-NTCP细胞中NF-κB的表达水平;进一步,过表达NF-κB显著上调HBV感染的HepG2-NTCP细胞中HBV RNA、HBV DNA、HBsAg和HBeAg水平,沉默时则相反;同时,双萤光素酶报告基因实验显示NF-κB可增强HBV Sp2启动子活性。回补实验发现多黏菌素B调控HBV转录复制依赖于NF-κB。结论:多黏菌素B通过下调NF-κB进而抑制HBV Sp2启动子活性,最终抑制HBV转录复制。
文摘OBJECTIVE Liver fibrosis is a chronic damage process related to the further progression of hepatic cirrhosis and has yet no truly effective treatment is available.This study aimed to investigate the effects of isochlorogenic acid A(ICQA)on liver fibrosis induced by carbon tetrachloride(CCl4)and clarify the underlying mechanism.METHODS Rats were treated with CCl4 for eight weeks in order to induce liver fibrosis and simultaneously orally administered with ICQA(10,20 and 40 mg·kg-1).RESULTS ICQA had significant protective effect on liver injury,inflammation,and fibrosis in rats.Meanwhile,ICQA prevented the activation of hepatic stellate cells(HSC)as indicated by inhibiting the overexpres⁃sion of a-smooth muscle actin(a-SMA).In addition,reduced fibrosis was found to be associated with decreased protein expression of high-mobility group box 1(HMGB1)and toll like receptor(TLR)4.Moreover,ICQA supressed the cytoplasmic translocation of HMGB1 in rat liver.Further investigations indicated that ICQA treatment significantly attenuated nuclear translocation of the nuclear factor-κB(NF-κB)p65 and inhibited degradation of IkBa expression in the liver of rats with liver fibrosis.CONCLUSION ICQA has hepatoprotective and anti-fibrotic effects in rats with liver fibrosis through modulating the HMGB1/TLR4/NF-κB signaling pathways.
