白藜芦醇通过巨噬细胞源性外泌体调节肝星状细胞活化对肝纤维化的影响  

Effect of resveratrol on liver fibrosis through regulating hepatic stellate cell activation by macrophage-derived extracellular vesicles

作　　者：梁耀丹 井洁[1] 张泰程 姜海行[1] 覃山羽[1] Liang Yaodan;Jing Jie;Zhang Taicheng;Jiang Haixing;Qin Shanyu(Department of Gastroenterology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院消化内科,南宁530021

出　　处：《广西医科大学学报》2023年第12期1949-1955,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81960119);广西壮族自治区卫生健康委自筹经费科研课题资助项目(No.Z-A20230546)。

摘　　要：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 ofα-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 ofα-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.

关 键 词：白藜芦醇 肝纤维化 外泌体 巨噬细胞 肝星状细胞 自噬 

分 类 号：R575.2[医药卫生—消化系统]