miR-203a-5p通过抑制丝裂原活化蛋白激酶1的表达调控人牙周膜干细胞的炎性反应

目的检测炎性人牙周膜干细胞(hPDLSCs)中miR-203a-5p及丝裂原活化蛋白激酶1(MAPK1)的表达水平,探讨二者间的靶向关系对牙周炎炎性反应的调控作用。方法分离培养及流式细胞计量术鉴定hPDLSCs;将hPDLSCs分为:对照组、LPS刺激组、miR-203a-5p模拟物转染组及miR-203a-5p抑制剂转染组,用RT-qPCR检测miR-203a-5p及MAPK1mRNA表达,Westernblot检测MAPK1蛋白表达,双荧光报告系统验证miR-203a-5p与MAPK1的靶向关系。结果干细胞表面标志物CD73及CD90阳性;LPS刺激组miR-203a-5p的表达明显上调(P<0.05);miR-203a-5p模拟物及LPS使miR-203a-5p的表达明显上调(P<0.05),同时MAPK1mRNA及蛋白的表达下调(P<0.05);miR-203a-5p抑制物使miR-203a-5p的表达量下降(P<0.05),同时MAPK1mRNA及蛋白的表达增加(P<0.05);miR-203a-5p与MAPK1具有靶向作用。结论miR-203a-5p通过靶向下调MAPK1基因的表达来调控牙周炎的炎性反应过程。

miR-203a-5p regulates the inflammation response of human periodontal membrane stem cells by inhibiting the expression of MAPK1

Objective To detect the expression of miR-203 a-5 p and MAPK1 in inflammation human periodontal ligament cells,and investigate the regulatory effects of miR-203 a-5 p and MAPK1 on periodontitis. Methods To isolate and identify of hPDLSCs by flow cytometry. HPDLSCs were divided into control group, LPS stimulation group, mir-203 a-5 p mimic transfection group and mir-203 a-5 p inhibitor transfection group. hPDLSCs were divided into the control group,LPS stimulation group, miR-203 a-5 p mimic transfected cells group and miR-203 a-5 p inhibitor transfection cells group. The effect of miR-203 a-5 p on MAPK1 gene was verified by RT-qPCR,Western blot and the dual-luciferase reporter assay. Results The expression of CD73 and CD90 was positive in hPDLSCs.The expression of miR-203 a-5 p significantly increased in LPS stimulation group(P<0.05). miR-203 a-5 p mimic and LPS could promote the expression of miR-203 a-5 p and suppress the expression of mRNA and protein of MAPK1(P<0.05);miR-203 a-5 p inhibitor could down-expression of miR-203 a-5 p and the expression of mRNA and protein of MAPK1 increased(P<0.05);luciferase assay showed that miR-203 a-5 p and MAPK1 had targeted comple-mentaryrelationship.Conclusions miR-203 a-5 p can regulate the inflammatory reaction process of periodontitis target by downregulating the expression of MAPK1 gene.