细胞外囊泡通过促进M1型巨噬细胞活化加重NASH进展  

Extracellular vesicles aggravate NASH progression by promoting the activation of M1 macrophages

作　　者：万志平 杨小安[1] 邓洪[1] Wan Zhiping;Yang Xiaoan;Deng Hong(Department of Infectious Diseases,the Third affiliated Hospital of Sun Yat-sen University,Guangdong Provincial Key Laboratory of Liver Disease Research,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院感染性疾病科,广东省肝脏疾病研究重点实验室,广州510630

出　　处：《新医学》2022年第7期484-490,共7页Journal of New Medicine

基　　金：国家自然科学基金(81870597)。

摘　　要：目的探讨脂肪变性肝细胞来源的细胞外囊泡(EV)对巨噬细胞活化的影响。方法选择SD大鼠予高脂肪高胆固醇(HFHCD)饮食构建非酒精性脂肪性肝炎(NASH)模型。用HE、油红O和Masson染色观察大鼠肝脏病变情况,免疫荧光染色检测肝脏M1型巨噬细胞活化。用棕榈酸(PA)诱导HepG2细胞为脂肪变性肝细胞,并用超速离心法提取其分泌的EV后进行鉴定,再用定量逆转录PCR(RT-qPCR)和流式细胞术检测EV对巨噬细胞的影响。结果大鼠肝组织HE、油红O和Masson染色结果显示NASH动物模型构建成功,免疫荧光染色检查证实HFHCD组肝脏CD68+CD86+细胞比例升高(P<0.05)。RT-qPCR显示PA组EV能促进M1型巨噬细胞标志物(IL-1β和IL-18)表达升高(P<0.001),流式细胞术显示PA组EV促进CD86+细胞比例增加(P<0.001),而EV抑制剂(GW4869)能阻断这种促进作用。结论脂肪变性肝细胞可通过EV促进M1型巨噬细胞活化加重NASH进展。Objective To explore the effect of extracellular vesicles(EVs)secreted by steatotic hepatocytes on the activation of macrophages.Methods The nonalcoholic steatohepatitis(NASH)model was established by feeding SD rats with high-fat and high-cholesterol diet(HFHCD).Hepatic lesions of rats were observed by HE,Oil Red O and Masson staining,and the activation of M1 macrophages in the liver was detected by immunofluorescent staining.HepG2 cells were induced to be steatotic hepatocytes with palmitic acid(PA),and their secreted EVs were isolated by ultracentrifugation.Then the effect of EVs on macrophages was observed by co-culture experiments and corresponding indicators were analyzed by RT-qPCR and flow cytometry.Results The results of HE,Oil Red O and Masson staining showed that the NASH animal model was successfully established.Immunofluorescent staining confirmed that the proportion of CD68+CD86+cells in the liver of HFHCD group was significantly increased(P<0.05).RT-qPCR showed that EVs in the PA group could up-regulate the expression of M1 macrophage markers(IL-1β,IL-18)(P<0.001).Flow cytometry demonstrated that EVs increased the proportion of CD86+cells in the PA group(P<0.001).EV inhibitors(GW4869)could block this promoting effect.Conclusion Steatotic hepatocytes can aggravate NASH progression by promoting the activation of M1 macrophages via EVs.

关 键 词：脂肪变性肝细胞 细胞外囊泡 M1型巨噬细胞 

分 类 号：R575.5[医药卫生—消化系统]