MicroRNA-146a对急性胰腺炎胰腺腺泡细胞增殖及凋亡的影响及其机制研究

The effects and mechanisms of microRNA-146a in the proliferation and apoptosis of pancreatic acinar cells

摘要:

目的 探讨microRNA-146a（miR-146a）对急性胰腺炎胰腺腺泡细胞增殖及凋亡的影响，并分析相关机制。方法 体外诱导并培养急性胰腺炎MPC-83细胞，将急性胰腺炎MPC-83细胞分为阴性对照组（mimics-NC组）、过表达miR-146a组（miR-146a-mimics组），另以未经诱导处理的MPC-83细胞为空白对照组（NG组）。采用实时荧光定量聚合酶链反应检测各组MPC-83细胞miR-146a，MTT法检测细胞增殖情况，流式细胞仪检测细胞凋亡情况，酶联免疫吸附试验检测肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6），Western blotting检测增殖细胞核抗原（PCNA）、Bcl-2相关X蛋白（Bax）、B淋巴细胞瘤2（Bcl-2）、核转录因子-κB p65（NF-кB p65）蛋白的表达。结果 急性胰腺炎细胞模型建立成功，成功转染后，与NG组、mimics-NC组比较，miR-146a-mimics组细胞凋亡率，TNF-α、IL-6、Bax和NF-κB p65蛋白相对表达量降低（P <0.05），细胞存活率、PCNA和Bcl-2蛋白相对表达量升高（P <0.05）。结论 过表达miRNA-146a可抑制急性胰腺炎MPC-83细胞凋亡并促进细胞增殖，其作用可能通过抑制NF-кB通路活化来实现。

Abstract:

Objective To investigate the effects of microRNA-146a (miR-146a) on the proliferation and apoptosis of pancreatic acinar cells in acute pancreatitis, and to analyze the related mechanisms.Methods MPC-83 cells were induced as acute pancreatitis models and cultured in vitro, and were divided into negative control group (mimics-NC group) and over-expression miR-146a-mimics group (miR-146a-mimics group). The uninduced MPC-83 cells were set as the blank control group (NG group). The level of miR-146a in MPC-83 cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The cell proliferation was detected by MTT assay, while the cell apoptosis was detected by flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6). The protein expressions of proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and nuclear factor kappa B (NF-κB) p65 were detected via Western blotting.Results Acute pancreatitis cell models were established successfully. After transfection, the cell apoptosis rate, the levels of TNF-α and IL-6, and the protein expressions of Bax and NF-κB p65 in the miR-146a-mimics group were lower than those in the NG group and the mimics-NC group (P < 0.05). In contrast, the cell viability and the relative protein expressions of PCNA and Bcl-2 were increased in the miR-146a-mimics group compared with those in the NG group and the mimics-NC group (P < 0.05).Conclusions Overexpression of miR-146a inhibits the apoptosis of acute pancreatitis MPC-83 cells but promotes their proliferation, possibly by suppressing the activation of NF-κB pathway.