摘要
目的:探讨核受体亚家族1 D组成员1(NR1D1)在肺动脉平滑肌细胞(PASMCs)增殖和迁移中的作用及机制。方法:使用Nr1d1过表达腺病毒(Ad-Nr1d1)外源性转染PASMCs,然后通过Western blot实验分析各组NR1D1、哺乳动物雷帕霉素靶蛋白复合体1(mTORC1)的下游经典蛋白S6和4EBP1的磷酸化改变,使用Ki-67免疫荧光染色分析细胞增殖能力改变,使用Transwell实验分析细胞迁移能力改变。通过胰岛素恢复降低的mTORC1活性后,进一步分析各组细胞增殖、迁移能力变化是否被逆转。结果:体外Ad-Nr1d1转染PASMCs后,NR1D1表达明显上调,低氧所致的PASMCs增殖、迁移和S6、4EBP1磷酸化都明显受到抑制。而利用胰岛素恢复降低的mTORC1活性后,过表达Nr1d1对PASMCs增殖和迁移的抑制作用明显减弱。结论:NR1D1可通过降低mTORC1活性抑制低氧所致PASMCs异常增殖和迁移。
AIM:To investigate the role and mechanism of nuclear receptor subfamily 1 group D member 1(NR1D1)in the proliferation and migration of pulmonary arterial smooth muscle cells(PASMCs).METHODS:NR1D1 was exogenously overexpressed in PASMCs through adenovirus carrying Nr1d1(Ad-Nr1d1)transfection.Western blot was performed to assess the protein expression of NR1D1 and the phosphorylation of classical mammaliant target of rapamycin complex 1(mTORC1)downstream factors,S6 and 4E-binding protein 1(4EBP1).Immunofluorescence staining for Ki-67 and Transwell assay were performed to assess cell proliferation and migration,respectively.After mTORC1 activity was restored by insulin,the proliferation and migration of PASMCs were further assessed in each group.RESULTS:Transfec-tion of PASMCs with Ad-Nr1d1 significantly increased the protein expression of NR1D1 in vitro.Overexpression of NR1D1 attenuated the proliferation and migration of hypoxia-challenged PASMCs.In addition,it suppressed the phosphorylation of S6 and 4EBP1 in hypoxia-challenged PASMCs.Insulin-induced restoration of mTORC1 activity attenuated the Ad-Nr1d1-mediated inhibition of cell proliferation and migration.CONCLUSION:NR1D1 suppresses the proliferation and migration of hypoxia-challenged PASMCs by reducing mTORC1 activity.
作者
白焰平
王明亮
甘雪晴
孙雄山
赵嘉琪
陈杰
BAI Yanping;WANG Mingliang;GAN Xueqing;SUN Xiongshan;ZHAO Jiaqi;CHEN Jie(Burn Ward,General Hospital of Western Theater Command,Chengdu 610083,China;Department of Cardiovascular Medicine,General Hospital of Western Theater Command,Chengdu 610083,China;School of Clinical Medicine,South-west Medical University,Luzhou 646000,China;Department of Cardiac Surgery,General Hospital of Western Theater Command,Chengdu 610083,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2023年第9期1605-1610,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.82100419)。
关键词
NR1D1蛋白
肺动脉平滑肌细胞
细胞增殖
细胞迁移
NR1D1 protein
pulmonary arterial smooth mucle cells
cell proliferation
cell migration
