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当归多糖通过调控miR-301a-5p介导人牙周膜干细胞增殖和凋亡
作者:李生婷  姚毅章  赵国廷 
单位:青海省第五人民医院 口腔医学中心, 青海 西宁 810007
关键词:当归多糖 miR-301a-5p 人牙周膜干细胞 增殖 凋亡 
分类号:R781.42;R285.5
出版年·卷·期(页码):2022·50·第三期(315-320)
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-301a-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-301a-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-301a-5p was increased significantly(P<0.001). Knockdown of miR-301a-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-301a-5p expression.

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