miR-423-5p靶向水通道蛋白1对雨蛙肽诱导的急性胰腺炎AR42J细胞损伤的影响

Effect of miR-423-5p on the injury of AR42J cells induced by caerulein through targeting AQP1

目的:研究miR-423-5p对雨蛙肽诱导的胰腺AR42J细胞损伤的影响及其分子作用机制。方法:通过雨蛙肽处理AR42J细胞建立胰腺炎损伤模型,qRT-PCR检测AR42J细胞中miR-423-5p的表达,Western blot检测水通道蛋白1(aquaporin 1,AQP1)、Bcl-2、Bax和caspase-3蛋白表达,ELISA法检测培养上清中IL-6和TNF-α的分泌水平,流式细胞术测定细胞凋亡率,双荧光素酶报告系统验证miR-423-5p与AQP1间的调控关系。结果:在雨蛙肽诱导的AR42J细胞中miR-423-5p表达上调,AQP1表达下调;抑制miR-423-5p和过表达AQP1均可减轻雨蛙肽诱导的AR42J细胞损伤,抑制细胞凋亡;双荧光素酶报告系统结果显示miR-423-5p靶向负调控AQP1的表达;抑制AQP1逆转了抑制miR-423-5p对雨蛙肽诱导的胰腺AR42J细胞损伤的作用。结论:miR-423-5p通过靶向AQP1减轻雨蛙肽诱导的胰腺AR42J细胞损伤,抑制细胞凋亡。

AIM:To investigate the effect of miR-423-5p on the injury of pancreatic AR42J cells induced by caerulein and explore the underlying molecular mechanism.METHODS:A model of pancreatitis injury was established by treating AR42J cells with caerulein.The expression level of miR-423-5p in AR42J cells was detected by qRT-PCR.The expression levels of aquaporin 1(AQP1),Bcl-2,Bax and capase3 were measured by Western blot.The levels of IL-6 and TNF-αin culture supernatants were detected by enzyme-linked immunosorbent assay(ELISA).The cell apoptotic rate was determined by flow cytometry.The relationship between miR-423-5p and AQP1 was verified by dual-luciferase reporter assay system.RESULTS:miR-423-5p was up-regulated and AQP1 was down-regulated in AR42J cells induced by caerulein.Inhibition of miR-423-5p or overexpression of AQP1 alleviated the injury of AR42J cells induced by caerulein and inhibited cell apoptosis.The results of dual-luciferase reporter assay system suggested that miR-423-5p targeted and negatively regulated the expression of AQP1.Inhibition of AQP1 reversed the effects of inhibition of miR-423-5p on the injury of AR42J cells induced by caerulein.CONCLUSION:miR-423-5p alleviates the injury of pancreatic AR42J cells induced by caerulein and inhibited apoptosis by targeting AQP1.