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-现代医学

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

单位：都江堰市人民医院骨科, 四川都江堰 611830

分类号：R329

出版年·卷·期（页码）：2022·50·第八期（929-933）

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hFOB 1.19，分为Con组(5.5 mmol·L^-1葡萄糖)、HG组(25.0 mmol·L^-1葡萄糖)、HG+miR-NC组(转染miR-NC+25.0 mmol·L^-1葡萄糖)、HG+miR-378组(转染miR-378模拟物+25.0 mmol·L^-1葡萄糖)、HG+SB203580组(10 μm SB203580+25.0 mmol·L^-1葡萄糖)、HG+miR-378+SB203580组(转染miR-378模拟物+10 μm SB203580+25.0 mmol·L^-1葡萄糖)。采用RT-qPCR检测miR-378、Runx2 mRNA、OCN mRNA表达量；采用MTT法检测细胞增殖；采用Western blot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、OCN mRNA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^-1 glucose),HG+SB203580 group (10 μm SB203580 and 25.0 mmol·L^-1 glucose),and HG+miR-378+SB203580 group (transfected with miR-378 mimic+10 μm 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 HG+miR-NC group was significantly down-regulated as compared with the Con group (P<0.05).The expression of p-p38MAPK protein 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,Ki67 protein expression,and mRNA expression levels of Runx2 and OCN in the HG+miR-378+SB203580 group were significantly down-regulated,and Cleaved-caspase3 protein expression was significantly up-regulated as compared with the HG+miR-378 group (P<0.05).Conclusion: The above results indicate that miR-378 promotes the proliferation and differentiation of osteoblasts under high-glucose conditions by activating the p38MAPK signaling pathway.

参考文献：

[1] ASPRY T J,HILL T R.Osteoporosis and the ageing skeleton[J].Subcell Biochem,2019,91(1):453-476.
[2] GENG Q,GAO H,YANG R,et al.Pyrroloquinoline quinone prevents estrogen deficiency-induced osteoporosis by inhibiting oxidative stress and osteocyte senescence[J].Int J Biol Sci,2019,15(1):58-68.
[3] 朱琦琦,戴芳芳.糖尿病性骨质疏松症的中医研究进展[J].中国骨质疏松杂志,2019,25(9):1331-1335.
[4] 田文青,周波.糖尿病性骨质疏松的研究进展[J].大医生,2018,3(8):110-111.
[5] 潘虹旭.MiRNA对成骨分化调控及骨质疏松发生的研究进展[J].中国骨质疏松杂志,2018,24(1):121-124.
[6] 姜徽,谢兴文,李宁.MicroRNA调控成骨分化与骨质疏松症相关性研究进展[J].中国骨质疏松杂志,2014(3):338-342.
[7] 龚一昕.淫羊藿次苷Ⅱ通过p38MAPK信号通路调控成骨细胞护骨素表达和成骨分化研究[D].合肥:安徽医科大学,2016.
[8] 刘家寅,刘光源,田发明,等.辛伐他汀通过p38MAPK信号通路诱导BMSCs成骨分化的研究[J].中国骨质疏松杂志,2016,22(2):125-130.
[9] 张璐瑶.2型糖尿病合并骨质疏松的相关因素分析[D].长春:吉林大学,2020.
[10] 赵明月,高永红.老年糖尿病患者饮食对骨质疏松症发生的影响[J].中国现代医药杂志,2020,22(2):53-55.
[11] 游利.miR-378对高糖作用下成骨细胞分化的调控作用及机制探讨[D].上海:上海交通大学,2015.
[12] 王俊成.miR-467f对高糖环境下小鼠骨髓间充质干细胞成骨分化的调节机制研究[D].北京:中国人民解放军医学院,2013.
[13] 于晓华,刘树泰,董凯,等.P38MAPK信号通路抑制剂SB203580对高糖状态下成骨细胞MC3T3-E1增殖和分化的影响[J].实用口腔医学杂志,2015,31(2):184-187.
[14] 朱晓峰,王廷春,张荣华,等.骨碎补总黄酮对高糖作用下成骨细胞分化及ERK_(1/2)和p38蛋白激酶表达的影响[J].中药材,2012,35(3):424-429.
[15] 钟海波,郭祥,黄琳惠.葛根素通过ERK1/2和p38 MAPK信号通路刺激成骨分化和骨形成的机制[J].中国比较医学杂志,2019,29(2):84-89.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录