MicroRNA-155调节Notch信号通路对TLCs诱导的急性胰腺炎腺泡细胞的影响

Effect of microRNA-155 on acute pancreatitis cells induced by TLCs through Notch signaling pathway

目的研究miRNA-155对TLCs诱导的胰腺AR42J细胞损伤的作用及其可能的分子机制。方法利用TLCs诱导AR42J细胞建立急性胰腺炎细胞模型,细胞分为6组:正常对照组,TLCs模型组,TLCs+microRNA-155 inhibitor组,TLCs+microRNA-155 negative control组,TLCs+microRNA-155 inhibitor+pcDNA negative control组,TLCs+microRNA-155 inhibitor+pcDNA-Nocth1组。利用RT-qPCR检测细胞中miRNA-155与Notch1mRNA的表达,利用western blot检测细胞中Notch1蛋白及凋亡相关蛋白Bcl-2,Bax,caspase-3的表达,用ELISA法检测细胞培养液中炎性因子IL-6,IL-32,IL-1β及TNF-α的含量。结果结果显示,与正常对照组相比较,模型组细胞中miRNA-155与Notch1 mRNA的表达均显著升高;与模型组相比较,抑制miRNA-155的表达后,TLCs+miRNA-155 inhibitor组细胞中Notch1的表达显著降低,抗凋亡蛋白Bcl-2的表达显著下降,凋亡蛋白Bax与Caspase-3的表达显著升高,炎性因子IL-6,IL-32,IL-1β及TNF-α的含量显著降低;与TLCs+miRNA-155 inhibitor相比较,TLCs+microRNA-155 inhibitor+pcDNA-Notch1组,抗凋亡蛋白Bcl-2的表达显著升高,凋亡蛋白Bax与Caspase-3的表达显著降低,炎性因子IL-6,IL-32,IL-1β及TNF-α的含量显著升高。结论通过抑制miRNA-155的表达可抑制Notch信号通路的激活,促进细胞凋亡,抑制炎性因子的释放,减轻细胞损伤,进而发挥细胞保护作用。

Objective To investigate effects of miRNA-155 on pancreatic AR42J cells injury induced by TLCs and its possible molecular mechanism.Methods Acute pancreatitis cell model was established by TLCs.AR42J cells were cultured and divided into 6 groups:normal control group,model group,TLCs+microRNA-155 inhibitor group,TLCs+microRNA-155 NC group,TLCs+microRNA-155 inhibitor+pcDNA NC group,TLCs+microRNA-155 inhibitor+pcDNA-Nocth1 group.RT-qPCR was used to detect the expression of miRNA-155 and Notch1 mRNA.Western blot was used to detect the expression of Notch1 protein,Bcl-2 protein,Bax protein and caspase-3 protein.ELISA was used to detect the contents of IL-6,IL-32,IL-1βand TNF-αin the cell culture medium.Results The results showed that the expression of miRNA-155 mRNA and Notch1 mRNA increased significantly in model group cells compared with normal control group cells.After miRNA-155 inhibition,the expression of Notch1 protein,and Bcl-2 protein decreased significantly,and the contents of IL-6,IL-32,IL-1βand TNF-αdecreased significantly TLCs+miRNA-155 inhibitor group cells,but the expression of Bax protien and caspase3 protein increased significantly,compared with the model group cells.Compared with TLCs+miRNA-155 inhibitor group,the expression of Bcl-2 protien and the contents of IL-6,IL-32,IL-1βand TNF-αincreased significantly,but the expression of Bax and caspase3 decreased significantly in TLCs+microRNA-155 inhibitor+pcDNA-Notch1group cells.Conclusion Inhibiting the expression of miRNA-155 can inhibit the activation of the Notch signaling pathway,promote cell apoptosis,suppress the release of inflammatory factors,reduce cell damage,and exert a cell-protective effect.