摘要
目的:探讨当归多糖(APS)调控miR-301a-5p对人牙周膜干细胞增殖和凋亡的影响。方法:将人牙周膜干细胞分为对照(Control)组、APS组、APS+anti-NC组、APS+anti-miR-301a-5p组。细胞计数试剂盒8(CCK-8)检测细胞活力;平板克隆形成实验检测细胞克隆形成能力;蛋白质印迹(Western blot)法检测蛋白表达;流式细胞术检测细胞凋亡;实时荧光定量PCR(RT-qPCR)检测miR-301a-5p表达水平;采用酶标法检测caspase3活性。结果:APS处理后人牙周膜干细胞的细胞活力显著升高,细胞克隆形成数增多,细胞凋亡率显著降低,增殖相关蛋白Ki-67、PCNA表达水平显著升高,抑凋亡相关蛋白Bcl-2表达水平显著升高,而促凋亡相关蛋白Bax、caspase3表达水平显著降低,caspase3活性降低,miR-301a-5p表达水平显著升高(P<0.001)。敲减miR-301a-5p逆转了APS对人牙周膜干细胞增殖和凋亡的作用。结论:APS通过上调miR-301a-5p表达促进人牙周膜干细胞增殖,抑制细胞凋亡。
Objective:To investigate the effect of angelica polysaccharide(APS)on the proliferation and apoptosis of human periodontal ligament stem cells.Methods:Human periodontal ligament stem cells were divided into control group,APS group,APS+anti-NC group,and APS+anti-miR-301 a-5p group.Cell counting kit 8(CCK-8)was used to detect cell viability;Plate colony formation assay was used to detect the ability of cells to form colonies.Western blot was used to detect protein expression;flow cytometry was used to detect apoptosis;real-time fluorescent quantitative PCR(RT-qPCR)was used to detect miR-301 a-5p expression levels.The activity of caspase3 was detected by enzyme labeling method.Results:After APS treatment,the cell viability of human periodontal ligament stem cells was increased significantly,the apoptosis rate was reduced significantly,the expression levels of proliferation-related proteins Ki-67 and PCNA were increased,and the expression levels of anti-apoptosis related protein Bcl-2 were increased significantly,while while the expression levels of pro-apoptosis-related protein Bax,caspase3 were reduced significantly,the activity of caspase3 was decreased,and the expression of miR-301 a-5p was increased significantly(P<0.001).Knockdown of miR-301 a-5p reversed the effects of Angelica polysaccharides on the proliferation and apoptosis of human periodontal ligament stem cells.Conclusion:Angelica polysaccharide can promote human periodontal ligament stem cells proliferation and inhibit apoptosis by up-regulating miR-301 a-5p expression.
作者
李生婷
姚毅章
赵国廷
LI Shengting;YAO Yizhang;ZHAO Guoting(Stomatological Center,the Fifth People’s Hospital of Qinghai Province,Xining 810007,China)
出处
《现代医学》
2022年第3期315-320,共6页
Modern Medical Journal
关键词
当归多糖
miR-301a-5p
人牙周膜干细胞
增殖
凋亡
angelica polysaccharide
miR-301a-5p
human periodontal ligament stem cells
proliferation
apoptosis
