非酒精性脂肪肝肝细胞来源外泌体中LncRNA MALAT1加剧胰腺细胞炎性反应的机制研究  

Mechanism Study of LncRNA MALAT1 in Hepatocyte-derived Exosomes in Nonalcoholic Fatty Liver Aggravates Pancreatic Inflammatory

作　　者：刘迪[1] 柳科军 侯绍章[2] LIU Di;LIU Kejun;HOU Shaozhang(Department of Hepatobiliary Surgery,General Hospital of Ningxia Medical University,First Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China;Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院肝胆外科,宁夏医科大学第一临床医学院,银川750004 [2]宁夏医科大学,银川750004

出　　处：《宁夏医科大学学报》2024年第12期1201-1207,共7页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金项目(2023AAC03581)。

摘　　要：目的 探讨非酒精性脂肪肝(NAFLD)肝细胞分泌的外泌体中LncRNA MALAT1对胰腺细胞炎性反应的影响及其机制。方法 用游离脂肪酸(FFA)(含有体积比为2∶1的油酸和棕榈酸)处理人肝细胞株HepG224 h建立非酒精性脂肪肝肝细胞模型,采用超速离心法提取外泌体并通过Western blot检测外泌体标志蛋白CD9、CD63、CD81和CYP2E1的表达水平;RT-qPCR检测MALAT1的表达;随后将外泌体与胰腺细胞AR42J共培养,ELISA检测炎症标志物IL-6、TNF-α转录水平;通过Western blot检测YAP蛋白磷酸化及Hippo通路相关蛋白水平。结果 NAFLD肝细胞来源外泌体中LncRNA MALAT1呈高表达,外泌体共培养的胰腺细胞中MALAT1的水平升高(P<0.05)。与正常肝细胞来源的外泌体相比,NAFLD肝细胞来源的外泌体能够诱导胰腺细胞表达IL-6和TNF-α,降低YAP蛋白的磷酸化水平(P均<0.05)。沉默MALAT1可抑制Hippo-YAP通路关键蛋白表达水平,而YAP通路抑制剂CA3显著逆转了MALAT1诱导的IL-6和TNF-α水平的升高(P<0.05)。结论 NAFLD肝细胞来源外泌体中LncRNA MALAT1可以加剧胰腺细胞的炎性反应,其机制可能与其激活YAP蛋白磷酸化从而调控炎性反应相关。Objective To investigate the effect and mechanism of LncRNA MALAT1 in exosomes secreted by hepatocytes of nonalcoholic fatty liver disease(NAFLD)on the inflammatory response of pancreatic cells.Methods Human normal liver cell line HepG2 was treated with FFA(containing oleic acid and palmitic acid with a volume ratio of 2∶1)for 24 h to establish a non-alcoholic fatty liver liver cell model.Exosomes were extracted by high-speed centrifugation and the expression levels of exosome marker proteins CD9,CD63,CD81 and CYP2E1 were detected by Western blot.The expression of MALAT1 was detected by RT-qPCR.Subsequently,exosomes were co-cultured with pancreatic cells AR42J,and the transcription levels of inflammatory markers IL-6 and TNF-αwere detected by ELISA.YAP protein phosphorylation and Hippo pathway-related protein levels were detected by Western blot.Results LncRNA MALAT1 was highly expressed in exosomes derived from NAFLD hepatocytes,and the level of MALAT1 in pancreatic cells co-cultured with exosomes was significantly increased(P<0.05).Compared with normal hepatocyte-derived exosomes,NAFLD hepatocyte-derived exosomes could significantly induce the expression of IL-6 and TNF-αin pancreatic cells and reduce the phosphorylation level of YAP protein(P all<0.05).Silencing MALAT1 could inhibit the expression of key proteins in Hippo-YAP pathway,while YAP pathway inhibitor CA3 significantly reversed the increase of IL-6 and TNF-αlevels induced by MALAT1(P<0.05).Conclusion LncRNA MALAT1 in NAFLD hepatocyte-derived exosomes can aggravate the inflammatory response of pancreatic cells,and its mechanism may be related to the activation of YAP protein phosphorylation to regulate the inflammatory response.

关 键 词：非酒精性脂肪肝 外泌体 LncRNA MALAT1 炎性反应 

分 类 号：R575.5[医药卫生—消化系统]