摘要
目的:探讨LncRNA TMPO-AS1对成骨细胞分化、增殖的影响及其对Nrf2/Keapl信号通路的调控作用。方法:采用地塞米松(Dex)处理人成骨肉瘤细胞MG63建立骨质疏松症细胞模型(Dex组),分别将pcDNA-NC、pcDNA-TMPO-AS1、si-NC、siTMPO-AS1转染至MG63细胞,分别将pcDNA-NC、pcDNA-TMPO-AS1转染至MG63细胞后加入Dex处理24 h,同时将正常培养的细胞作为NC组;采用RT-qPCR法检测TMPO-AS1、ALP、RUNX2、OCN mRNA的表达水平;采用CCK-8法检测细胞增殖;采用Western blotting法检测CyclinD1、ALP、RUNX2、OCN、Nrf2、Keapl蛋白相对表达量。结果:与NC组比较,Dex组TMPOAS1的表达水平降低(P<0.05),OD值降低(P<0.05),CyclinD1、ALP、RUNX2、OCN、Nrf2、Keapl的表达水平降低(P<0.05);与pcDNA-NC组比较,pcDNA-TMPO-AS1组OD值升高(P<0.05),CyclinD1、ALP、RUNX2、OCN、Nrf2、Keapl蛋白水平升高(P<0.05),而转染si-TMPO-AS1的作用与之相反;与pcDNA-NC+Dex组比较,pcDNA-TMPO-AS1+Dex组OD值升高(P<0.05),CyclinD1、ALP、RUNX2、OCN、Nrf2、Keapl蛋白水平升高(P<0.05)。结论:TMPO-AS1过表达可能通过激活Nrf2/Keapl信号通路从而促进成骨细胞增殖及分化。
Objective:To investigate the effect of LncRNA TMPO-AS1 on the differentiation and proliferation of osteoblasts and its regulation of Nrf2/Keapl signaling pathway.Methods:Dexamethasone(Dex) was used to treat human osteosarcoma cells MG63 to establish an osteoporosis cell model(Dex group).The pcDNA-NC,pcDNA-TMPO-AS1,si-NC,and si-TMPO-AS1 were transfected into MG63 cells respectively.The pcDNA-NC and pcDNA-TMPO-AS1 were transfected into MG63 cells respectively and treated with Dex for 24 h.while the normal cultured cells were regarded as the NC group.RT-qPCR was used to detect the expression levels of TMPOAS1,ALP,RUNX2,and OCN mRNA.CCK-8 method was employed to detect cell proliferation,and western blotting method was adopted to detect CyclinD1,ALP,RUNX2,OCN,Nrf2,and Keapl protein expression.Results:Compared with the NC group,the expression level of TMPO-AS1 in Dex group was decreased,the OD value was decreased,and the expression levels of CyclinD1,ALP,RUNX2,OCN,Nrf2,and Keapl were decreased(P<0.05).Compared with pcDNA-NC group,the OD value of pcDNA-TMPO-AS1 group was increased,the protein levels of CyclinD1,ALP,RUNX2,OCN,Nrf2,and Keapl were increased(P<0.05),while the transfection of si-TMPO-AS1 had the opposite effect.Compared with pcDNA-NC+Dex group,the OD value of pcDNATMPO-AS1+Dex group was increased,and the protein levels of CyclinD1,ALP,RUNX2,OCN,Nrf2,and Keapl were increased(P<0.05).Conclusion:The overexpression of TMPO-AS1 may promote the proliferation and differentiation of osteoblasts by activating Nrf2/Keapl signaling pathway.
作者
苏亚
李小凤
李辉
Su Ya;Li Xiaofeng;Li Hui(Department of Endocrinology,Shanxi Provincial People's Hospital,Xi'an 710068,China)
出处
《广西医科大学学报》
CAS
2022年第2期318-322,共5页
Journal of Guangxi Medical University
