哈巴苷通过抑制miR-1246表达对LPS诱导的人牙周膜细胞损伤的影响  

Harpagide inhibits expression of miR-1246 on LPS-induced damage to human periodontal ligament cells

作　　者：吴源[1] 高鹏[1] 李海涛[1] WU Yuan;GAO Peng;LI Hai-Tao(Department of Stomatology,Qinghai Red Cross Hospital,Xining 810000,China)

机构地区：[1]青海红十字医院口腔科,西宁810000

出　　处：《中国免疫学杂志》2021年第17期2099-2103,共5页Chinese Journal of Immunology

摘　　要：目的:探讨哈巴苷对LPS诱导的人牙周膜细胞(hPDLCs)损伤的影响及分子机制。方法:hPDLCs分为Con组、LPS组、哈巴苷低、中、高浓度组(LPS+HG-L、M、H)、LPS+anti-miR-NC组、LPS+anti-miR-1246组、LPS+HG+miR-NC组、LPS+HG+miR-1246组。ELISA检测TNF-α、IL-1β水平;流式细胞术检测细胞凋亡;Western blot法检测蛋白表达;实时荧光定量PCR(RT-qPCR)检测miR-1246表达水平。结果:LPS诱导的hPDLCs中TNF-α、IL-1β水平升高,细胞凋亡率升高,Bcl-2表达水平降低,Bax表达水平升高,miR-1246表达水平升高(P<0.05)。哈巴苷低、中、高浓度处理后,LPS诱导的hPDLCs中TNF-α、IL-1β水平降低,细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低,miR-1246表达水平降低,且呈浓度依赖性(P<0.05)。抑制miR-1246表达后,LPS诱导的hPDLCs中TNF-α、IL-1β水平降低,细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低(P<0.05)。miR-1246过表达逆转了哈巴苷对LPS诱导的hPDLCs损伤的作用。结论:哈巴苷通过抑制miR-1246表达保护hPDLCs免受LPS诱导的损伤。Objective:To explore the effect of harpagide on LPS-induced injury of human periodontal ligament cells(hPDLCs)and its molecular mechanism.Methods:hPDLCs were divided into Con group,LPS group,low,medium,and high concentration groups of harpagide(LPS+HG-L,M,H),LPS+anti-miR-NC group,LPS+anti-miR-1246 group,LPS+HG+miR-NC group,LPS+HG+miR-1246 group.ELISA was used to detect the levels of TNF-αand IL-1β;Flow cytometry was used to detect cell apoptosis;Western blot was used to detect protein expression;Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect miR-1246 expression.Results:In LPS-induced human periodontal ligament cells,the levels of TNF-αand IL-1βwere increased,cell apoptosis was increased,Bcl-2 expression was decreased,Bax expression was increased,and miR-1246 expression was increased(P<0.05).After treatment with low,medium and high concentrations of harpagide,the levels of TNF-αand IL-1βin hPDLCs induced by LPS were decreased,the rate of apoptosis was decreased,the expression of Bcl-2 was increased,the expression of Bax was decreased,and the expression of mi-1246 was decreased,and was concentration-dependent(P<0.05).After inhibiting the expression of miR-1246,the levels of TNF-αand IL-1βin hPDLCs induced by LPS were decreased,the apoptosis rate was decreased,the expression of Bcl-2 was increased,and the expression of Bax was decreased(P<0.05).The overexpression of miR-1246 reversed the effect of harpagide on LPS-induced injury of hPDLCs.Conclusion:hPDLCs from LPS-induced damage by inhibiting the expression of miR-1246.

关 键 词：哈巴苷 miR-1246 人牙周膜细胞 炎症因子 凋亡 损伤 

分 类 号：R781.42[医药卫生—口腔医学]