Jichu yixue yu linchuang (Jan 2024)
miR-142-3p inhibits cerulein-induced apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1
Abstract
Objective To investigate the effect of miR-142-3p on the apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1. Methods AR42J cells were divided into blank group (blank), acute pancreatitis model group (AP, 100 nmol/L cerulein for 24 h), and then transfected with miR-142-3p mimics, mimics NC,miR-142-3p inhibitor and inhibitor NC, respectively. The cells in the model group were recorded as miR-142-3p mimics group, mimics NC group, miR-142-3p inhibitor group and inhibitor NC.The expression of miR-142-3p in cells was detected by RT-qPCR. The protein expressions of HMGB1, caspase-3, Bax and Bcl-2 were detected by Western blot. Hoechst staining was used to determine cell apoptosis. The apoptosis rate of cells was detected by flow cytometry. The targeting relationship between miR-142-3p and Hmgb1 was determined by dual luciferase reporter gene assay. Results Compared with blank control group, the expression level of miR-142-3p in the AP group was significantly down-regulated (P< 0.01), the expression level of HMGB1 and caspase-3 proteins was up-regulated(P< 0.05), the expression level of Bax protein was significantly up-regulated (P< 0.01), the expression level of Bcl-2 protein was significantly decreased(P< 0.01) and the apoptosis rate increased significantly (P< 0.01). Compared with the mimics NC group, the level of miR-142-3p in the miR-142-3p mimics group was significantly up-regulated(P< 0.01), the expression of HMGB, caspase-3 and Bax proteins was significantly down-regulated (P< 0.01), the expression of Bcl-2 protein was up-regulated (P< 0.05), and the apoptosis rate decreased significantly (P< 0.01).Compared with inhibitor NC group, the expression level of miR-142-3p in miR-142-3p inhibitor group was down-regulated (P< 0.05), the expression levels of HMGB1, caspase-3 and Bax proteins were significantly up-regulated (P< 0.01), the expression level of Bcl-2 protein was decreased (P< 0.05) and the apoptosis rate increased significantly (P< 0.01) .The dual luciferase reporter gene assay showed that Hmgb1 was the target gene of miR-142-3p. Conclusions 1)The expression of miR-142-3p was low in the model group. 2)miR-142-3p can inhibit the apoptosis of AR42J cells by inhibiting the expression of Hmgb1.
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