lncRNA-TNFRSF13C调控miR-1246对牙周细胞低氧诱导因子1α的作用  

Effect of lncRNA-TNFRSF13C on hypoxia-inducible factor 1alpha in periodontal cells by modulation of miR-1246

作　　者：白静 张雪 任燕 李月辉 田晓宇 Bai Jing;Zhang Xue;Ren Yan;Li Yuehui;Tian Xiaoyu(Department of Stomatology,Tangshan Central Hospital,Tangshan 063000,Hebei Province,China;Tangshan Fengrun District People’s Hospital,Tangshan 063000,Hebei Province,China;Kailuan(Group)Co.,Ltd.Tangjiazhuang Hospital,Tangshan 063000,Hebei Province,China;Zunhua People’s Hospital,Zunhua 064200,Hebei Province,China)

机构地区：[1]唐山中心医院口腔科,河北省唐山市063000 [2]唐山市丰润区人民医院,河北省唐山市063000 [3]开滦(集团)有限责任公司唐家庄医院,河北省唐山市063000 [4]遵化市人民医院,河北省遵化市064200

出　　处：《中国组织工程研究》2025年第5期928-935,共8页Chinese Journal of Tissue Engineering Research

基　　金：河北省医学科学基金资助项目(20230240)。

摘　　要：背景:LncRNA-TNFRSF13C是B细胞发育和功能中的一个重要因子,在牙周炎患者牙周组织中高表达,但具体机制还不清楚。目的:探讨lncRNA-TNFRSF13C调控miR-1246对牙周细胞低氧诱导因子1α的作用机制。方法:人牙周膜细胞经过脂多糖处理后分为7组:A组(无转染的人牙周膜细胞株)、B组(人牙周膜细胞株转染TNFRSF13C NC-siRNA)、C组(人牙周膜细胞株转染TNFRSF13C-siRNA)、D组(人牙周膜细胞株转染miR-1246 mimics)、E组(人牙周膜细胞株转染miR-1246 siRNA)、F组(人牙周膜细胞株转染TNFRSF13C-siRNA+miR-1246 mimics)及G组(人牙周膜细胞株转染TNFRSF13C-siRNA+miR-1246 siRNA)。采用qRT-PCR检测各组细胞lncRNA-TNFRSF13C、miR-1246相对表达;CCK-8检测细胞活性;流式细胞仪检测细胞凋亡率;免疫印迹检测细胞中低氧诱导因子1α、血管内皮生长因子蛋白的表达;Pearson分析lncRNA-TNFRSF13C与miR-1246相关性,双荧光素酶分析靶向关系。结果与结论:①A、B两组人牙周膜细胞活性、凋亡率及蛋白指标比较差异均无显著性意义(P>0.05),与B组相比,C组细胞活性升高、凋亡率及低氧诱导因子1α、血管内皮生长因子蛋白降低(P<0.05),与C组相比,D组细胞活性降低,凋亡率及低氧诱导因子1α、血管内皮生长因子蛋白均升高(P<0.05),D组相比,E组细胞活性升高(P<0.05),F组细胞活性低于E组,E、F组细胞凋亡率降低(P<0.05),与F组相比,G组细胞活性升高、凋亡率及低氧诱导因子1α、血管内皮生长因子蛋白降低(P<0.05)。②lncRNA-TNFRSF13C与miR-1246呈现正相关(P<0.05)。③TNFRSF13C-siRNA组与TNFRSF13C-NC组在miR-1246-wt荧光活性降低(P<0.05);与miR-1246-NC组相比,miR-1246 mimics组低氧诱导因子1α-wt、血管内皮生长因子-wt荧光活性升高(P<0.05)。④结果说明,下调lncRNA-TNFRSF13C对脂多糖处理的牙周细胞具有促进活性、降低凋亡的作用,同时也可抑制低氧诱导因子1α、血管内皮生长因子的表达,其机制�BACKGROUND:LncRNA-TNFRSF13C,an important factor in B cell development and function,is expressed in periodontal tissues of patients with periodontitis,but the specific mechanism is still unclear.OBJECTIVE:To investigate the mechanism of lncRNA-TNFRSF13C regulating miR-1246 on hypoxia-inducible factor 1αin periodontal cells.METHODS:Human periodontal ligament cells(hPDLCs)were treated with lipopolysaccharide and divided into group A(hPDLCs cell lines without transfection),group B(hPDLCs cell lines transfected with TNFRSF13C NC-siRNA),group C(hPDLCs cell lines transfected with TNFRSF13C-siRNA),group D(hPDLCs cell line transfected with miR-1246 mimics),group E(hPDLCs cell line transfected with miR-1246 siRNA),group F(hPDLCs cell line transfected with TNFRSF13CsiRNA+miR-1246 mimics),and group G(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 siRNA).The relative expression of lncRNA-TNFRSF13C and miR-1246 in each group was detected by qRT-PCR.Cell counting kit-8 assay was used to detect cell viability.Apoptosis was detected by flow cytometry.Expression of hypoxia-inducible factor 1αand vascular endothelial growth factor proteins was detected by western blot.The correlation between lncRNATNFRSF13C and miR-1246 was analyzed by Pearson,and the targeting relationship was analyzed by dual-luciferase reporter assay.RESULTS AND CONCLUSION:There was no significant difference in human periodontal ligament cell activity,apoptosis rate and protein indexes between groups A and B(P>0.05).Compared with group B,hPDLCS cell activity in group C was increased,and apoptosis rate and the expression of hypoxia-inducible factor 1αand vascular endothelial growth factor proteins were decreased(P<0.05).Compared with group C,hPDLCS cell activity in group D was decreased,and apoptosis rate and the expression of hypoxia-inducible factor 1αand vascular endothelial growth factor proteins were increased(P<0.05).Compared with group D,the cell activity of group E was increased(P<0.05).The cell activity in group F was lower than that in g

关 键 词：牙周炎 牙周膜细胞 脂多糖 炎症 miR-1246 lncRNA-TNFRSF13C 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R781