miR-378通过p38MAPK信号通路对高糖作用下成骨细胞分化的影响及机制研究  

Effect of miR-378 on osteoblast differentiation under high-glucose conditions via p38MAPK signaling pathway and its mechanism

作　　者：马宽 徐德兴 银保[1] 熊科 MA Kuan;XU Dexing;YIN Bao;XIONG Ke(Department of Orthopedics,People's Hospital of Dujiangyan,Dujiangyan 611830,China)

机构地区：[1]都江堰市人民医院骨科,四川都江堰611830

出　　处：《现代医学》2022年第8期929-933,共5页Modern Medical Journal

摘　　要：目的:探究miR-378通过p38MAPK信号通路对高糖作用下成骨细胞分化的影响及机制。方法:培养成骨细胞hFOB1.19,分为Con组(5.5mmol·L^(-1)葡萄糖)、HG组(25.0mmol·L^(-1)葡萄糖)、HG+miR-NC组(转染miR-NC+25.0mmol·L^(-1)葡萄糖)、HG+miR-378组(转染miR-378模拟物+25.0mmol·L^(-1)葡萄糖)、HG+SB203580组(10μmSB203580+25.0mmol·L^(-1)葡萄糖)、HG+miR-378+SB203580组(转染miR-378模拟物+10μmSB203580+25.0mmol·L^(-1)葡萄糖)。采用RT-qPCR检测miR-378、Runx2mRNA、OCNmRNA表达量;采用MTT法检测细胞增殖;采用Westernblot法检测Ki67、Cleaved-caspase3、磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)蛋白表达。结果:与Con组比较,HG组和HG+miR-NC组miR-378表达量显著下调(P<0.05);与HG组和HG+miR-NC组比较,HG+miR-378组miR-378表达量显著上调(P<0.05)。与Con组比较,HG组和HG+miR-NC组细胞增殖能力、Ki67蛋白表达显著下调,Cleaved-caspase3蛋白显著上调(P<0.05);与HG组和HG+miR-NC组比较,HG+miR-378组细胞增殖能力、Ki67蛋白表达显著上调,Cleaved-caspase3蛋白显著下调(P<0.05)。与Con组比较,HG组和HG+miR-NC组Runx2、OCNmRNA表达显著下调(P<0.05);与HG组和HG+miR-NC组比较,HG+miR-378组Runx2、OCN mRNA表达显著上调(P<0.05)。与Con组比较,HG组和HG+miR-NC组p-p38MAPK蛋白表达显著下调(P<0.05);与HG组和HG+miR-NC组比较,HG+miR-378组p-p38MAPK蛋白表达显著上调(P<0.05)。与HG+miR-378组比较,HG+miR-378+SB203580组细胞增殖能力、Ki67蛋白表达及Runx2、OCN mRNA表达显著下调,Cleaved-caspase3蛋白显著上调(P<0.05)。结论:miR-378通过激活p38MAPK信号通路促进高糖作用下成骨细胞的增殖和分化。Objective:To investigate the effect of miR-378 on osteoblast differentiation under high-glucose conditions via p38MAPK signaling pathway and its mechanism.Methods:The human osteoblasts hFOB 1.19 were cultured and divided into Con group(5.5 mmol·L^(-1)glucose),HG group(25.0 mmol·L^(-1)glucose),HG+miRNC group(transfected with miR-NC and 25.0 mmol·L^(-1)glucose),HG+miR-378 group(transfected with miR-378 mimic and 25.0 mmol·L glucose),HG+SB203580 group(10μm SB203580 and 25.0 mmol·L^(-1)glucose),and HG+miR-378+SB203580 group(transfected with miR-378 mimic+10μm SB203580+25.0 mmol·L^(-1)glucose).RT-qPCR was used to detect the expression levels of miR-378,Runx2 mRNA and OCN mRNA.MTT assay was used to detect cell proliferation.Western blot was used to detect the protein expression of Ki67,Cleaved-caspase3 and phosphorylated p38 mitogen-activated protein kinase(p-p38MAPK).Results:The expression levels of miR-378 in the HG group and the HG+miR-NC group were significantly down-regulated as compared with the Con group(P<0.05).The expression level of miR-378 in the HG+miR-378 group was significantly up-regulated as compared with the HG group and the HG+miR-NC group(P<0.05).The proliferation ability and Ki67 protein expression were significantly down-regulated,and Cleaved-caspase3 protein expression was significantly up-regulated in the HG group and the HG+miR-NC group as compared with the Con group(P<0.05).The proliferation ability and Ki67 protein expression were significantly up-regulated,and Cleaved-caspase3 protein expression was significantly down-regulated in the HG+miR-378 group as compared with the HG group and the HG+miR-NC group(P<0.05).The mRNA expression levels of Runx2 and OCN in the HG group and the HG+miR-NC group were significantly down-regulated as compared with the Con group(P<0.05).The mRNA expression levels of Runx2 and OCN in the HG+miR-378 group were significantly up-regulated as compared with the HG group and the HG+miR-NC group(P<0.05).The expression of p-p38MAPK protein in the HG group and the

关 键 词：miR-378 P38MAPK信号通路 成骨细胞 分化 机制 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]