目的:探讨白藜芦醇(RSV)通过调节巨噬细胞影响肝星状细胞活化的机制.方法:以超速离心法提取空白组(NC-E组)和RSV处理(RSV-E)组外泌体并进行鉴定.将NC-E组及RSV-E组外泌体与转化生长因子-β1(TGF-β1)活化的肝星状细胞JS-1共培养,分为对照组、TGF-β1活化组(TGF-β1组)、NC-E+TGF-β1活化组(NC-E+TGF-β1组)和RSV-E+TGF-β1活化组(RSV-E+TGF-β1组).采用免疫荧光法、实时荧光定量PCR(RT-qPCR)、western blotting检测肌动蛋白A(α-SMA)和Ⅰ型胶原a1链(Col1a1)纤维化指标表达.Western blotting检测Beclin1和LC3Ⅰ/Ⅱ自噬相关指标表达.结果:与对照组相比,TGF-β1组α-SMA、Col1a1、Beclin1和LC3Ⅰ/Ⅱ表达水平升高(P<0.05);与TGF-β1组相比,RSV-E+TGF-β1组α-SMA、Col1a1、Beclin1和LC3Ⅰ/Ⅱ表达水平下降(P<0.05),NC-E+TGF-β1组与TGF-β1组差异无统计学意义(P>0.05).结论:RSV可能通过调节巨噬细胞源性外泌体抑制肝纤维化,其机制可能与抑制肝星状细胞自噬有关.
Objective:To investigate the mechanism of resveratrol(RSV)on the activation of hepatic stellate cells by regulating macrophages.Methods:Exosomes were extracted from blank(NC-E)group and resveratrol-treated(RSV-E)group by ultracentrifugation method,and the extracts were identified.The exosomes from the NC-E group and the RSV-E group were co-cultured with growth factor-β1(TGF-β1)-activated hepatic stellate cells JS-1 and divided into control group,TGF-β1-activated group(TGF-β1 group),NC-E+TGF-β1-activated group(NC-E+TGF-β1 group)and RSV-E+TGF-β1-activated group(RSV-E+TGF-β1 group).Immunofluores-cence,real-time fluorescence quantitative PCR(RT-qPCR)and western blotting were used to detect the alpha-smooth muscle actin(α-SMA)and type I collagen alpha 1 chain(Col1a1).Western blotting was also used to de-tect the expression of autophagy-related indicators Beclin1 and LC3Ⅰ/Ⅱ.Results:Compared with the control group,the TGF-β1 group showed increased expression levels of α-SMA,Col1a1,Beclin1 and LC3Ⅰ/Ⅱ(P<0.05).Compared with the TGF-β1 group,the RSV-E+TGF-β1 group showed decreased expression levels of α-SMA,Col1a1,Beclin1 and LC3Ⅰ/Ⅱ(P<0.05),while the NC-E+TGF-β1 group and TGF-β1 group showed no significant difference(P>0.05).Conclusion:RSV may inhibit the progression of liver fibrosis by regulating macrophage-derived extracellular vesicles and its mechanism may be related to the inhibition of autophagy in he-patic stellate cells.