过表达miR-21对脂多糖诱导人牙髓干细胞炎症、凋亡和自噬的影响  

Effects of overexpression of miR-21 on LPS-induced inflammation,apoptosis,and autophagy in hDPSCs

作　　者：林玉祥 林慧 车立纯 田河 LIN Yuxiang;LIN Hui;CHE Lichun;TIAN He(The Second Department of Cariology and Endodontology Stomatology Hospital of Xiamen Medical College,Xiamen 361000,China;不详)

机构地区：[1]厦门医学院附属口腔医院牙体牙髓病二科,福建厦门361000 [2]厦门市口腔疾病诊疗重点实验室 [3]青岛市中心医院口腔科

出　　处：《山东医药》2021年第23期33-36,共4页Shandong Medical Journal

基　　金：福建省厦门市科技局医疗卫生科技计划项目(3502Z20194078)。

摘　　要：目的观察过表达miR-21对脂多糖(LPS)诱导的人牙髓干细胞(hDPSCs)炎症、凋亡和自噬的影响。方法取生长良好的第3~4代hDPSCs,随机分为对照组、LPS组、LPS+miR-NC mimics组、LPS+miR-21 mimics组,LPS+miR-NC mimics组、LPS+miR-21 mimics组分别使用Lipofectamine 2000转染试剂转染miR-NC mimics、miR-21mimics,LPS组、LPS+miR-NC mimics组、LPS+miR-21 mimics组均加入5μg/mL LPS,继续培养48 h。采用ELISA法检测各组细胞上清液白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)水平,流式细胞术检测细胞凋亡率,Western blotting法检测凋亡相关蛋白裂解的半胱氨酸蛋白酶3(Cleaved Caspase-3)和自噬相关蛋白死骨片1(p62)、自噬微管相关蛋白轻链3抗体(LC3B)表达。结果与对照组比较,LPS组、LPS+miR-NC mimics组、LPS+miR-21 mimics组细胞上清液IL-1β、IL-6、TNF-α水平均升高,且LPS组、LPS+miR-NC mimics组升高更明显(P均<0.05)。与对照组比较,LPS组、LPS+miR-NC mimics组、LPS+miR-21 mimics组细胞凋亡率及细胞Cleaved Caspase-3、LC3BⅡ表达均升高,p62、LC3BⅠ表达均降低,且LPS组、LPS+miR-NC mimics组变化更明显(P均<0.05)。结论 LPS可诱导hDPSCs发生炎症反应,并促进其凋亡和自噬,而过表达miR-21可以抑制LPS的上述作用。Objective To investigate the effects of overexpression of miR-21 on lipopolysaccharide(LPS)-induced inflammation,apoptosis,and autophagy of human dental pulp stem cells(hDPSCs).Methods The hDPSCs of the third to fourth generations with good growth were randomly divided into the control group,LPS group,LPS+miR-NC mimics group,and LPS+miR-21 mimics group.Lipofectamine 2000 transfection reagent was used to transfect miR-NC mimics and miR-21 mimics into hDPSCs of the LPS+miR-NC mimics group and the LPS+miR-21 mimics group,respectively.LPS group,LPS+miR-NC mimics group and LPS+miR-21 mimics group were added with 5μg/mL LPS and then the culture was continued for 48 h.The expression levels of IL-1β,IL-6 and TNF-αin the supernatant of cells in each group were de-tected by ELISA.The apoptosis rate was detected by flow cytometry.The expression levels of apoptosis-related protein Cleaved Caspase-3 and autophagy-related protein p62,LC3B were detected by Western blotting.Results Compared with the Control group,the levels of IL-1β,IL-6 and TNF-αincreased in the cell supernatant of the LPS group,LPS+miR-NC mimics group,and LPS+miR-21 mimics group,and the increase was more significant in the LPS group and LPS+miR-NC mimics group(all P<0.05).Compared with the control group,the apoptosis rate and the levels of Cleaved Caspase-3 and LC3BⅡincreased in the LPS group,LPS+miR-NC mimics group and LPS+miR-21 mimics group,while the levels of p62 and LC3BⅠdecreased.Moreover,the changes in the LPS group and LPS+miR-NC mimics group were more significant(all P<0.05).Conclusion LPS can induce the inflammatory response of hDPSCs and promote their apoptosis and autophagy,while overexpression of miR-21 can inhibit these effects of LPS.

关 键 词：牙髓干细胞 MIR-21 脂多糖 细胞凋亡 细胞自噬 Cleaved Caspase-3 IL-1Β IL-6 TNF-α LC3B 

分 类 号：R78[医药卫生—口腔医学]