基于PI3K/AKT通路探究上调miR-210对牙髓干细胞增殖、凋亡能力的影响  

Effects of miR-210 on Proliferation and Apoptosis of Dental Pulp Stem Cells Based on PI3K/AKT Pathway

作　　者：于璐 余少华 熊宇 税桦桦 王迪 黄文静 YU Lu;YU Shao-hua;XIO NG Yu;SHUI Hua-hua;WANG Di;HUANG Wen-jing(Department of Stomatology,the First Affiliated Hospital of Army Medical University,Chongqing,400038,China)

机构地区：[1]陆军军医大学第一附属医院口腔科,重庆400038

出　　处：《现代生物医学进展》2021年第7期1212-1216,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81671022)。

摘　　要：目的:研究基于磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路探究上调微小RNA 210(miR-210)对大鼠牙髓干细胞增殖、凋亡能力的影响。方法:选取10只健康Sprague-Dawley(SD)雄性大鼠,颈椎脱臼处死后提取大鼠下切牙牙髓,进行牙髓干细胞培养和鉴定。分为正常组(未进行处理),mi R-210抑制组(给予20 nmol/L的mi R-210抑制物),mi R-210对照组(给予20 nmol/L的mi R-210模拟物)三组。采用CCK-8法检测牙髓干细胞增殖活性,酶联免疫吸附试验(ELISA)检测ALP活性,流式细胞仪检测细胞凋亡,采用免疫印迹(Western blot)检测PI3K、AKT蛋白。结果:与正常组相比,mi R-210抑制组细胞增殖、ALP活性降低,细胞凋亡率升高;mi R-210对照组细胞增殖、ALP活性升高,细胞凋亡率降低(P<0.05)。与mi R-210抑制组相比,mi R-210对照组细胞增殖、ALP活性升高,细胞凋亡率降低(P<0.05)。与正常组相比,mi R-210抑制组PI3K、p-AKT蛋白表达降低,mi R-210对照组PI3K、p-AKT蛋白表达升高(P<0.05)。与mi R-210抑制组相比,mi R-210对照组PI3K、p-AKT蛋白表达升高(P<0.05)。结论:mi R-210通过调控PI3K、p-AKT蛋白激活PI3K/AKT通路,促进大鼠牙髓干细胞增殖,抑制牙髓干细胞凋亡。Objective:To investigate the effect of microRNA 210(mir-210)on the proliferation and apoptosis of rat dental pulp stem cells based on phosphatidylinositol 3 kinase(PI3 K)/protein kinase B(Akt)pathway.Methods:Ten healthy Sprague Dawley(SD)male rats were selected.After cervical dislocation,the pulp of the lower incisors was extracted,and dental pulp stem cells were cultured and identified.They were divided into three groups:normal group(untreated),mir-210 inhibitory group(treated with 20 nmol/L mir-210 inhibitor),and mir-210 control group(treated with 20 nmol/L mir-210 mimic).CCK-8 method was used to detect the proliferation activity of dental pulp stem cells.Enzyme linked immunosorbent assay(ELISA)was used to detect ALP activity.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect PI3 K and AKT proteins.Results:Compared with the normal group,the proliferation and ALP activity of mir-210 group were decreased,while the apoptosis rate was increased in mir-210 control group(P<0.05).Compared with mir-210 group,mir-210 control group increased cell proliferation,ALP activity and decreased apoptosis rate(P<0.05).Compared with the normal group,the expression of PI3 K and p-AKT in mir-210 inhibition group was lower than that in mir-210 control group(P<0.05).Compared with mir-210 group,the expression of PI3 K and p-AKT protein in mir-210 control group was increased(P<0.05).Conclusion:Mi R-210 promotes the proliferation of rat dental pulp stem cells and inhibits the apoptosis of dental pulp stem cells by regulating PI3 K and p-AKT proteins and activating PI3 K/AKT pathway.

关 键 词：牙髓干细胞 增殖 凋亡 磷脂酰肌醇3激酶 蛋白激酶B 

分 类 号：R-33[医药卫生] R329.2