NR1D1介导低氧诱导的PASMCs异常增殖和迁移  

NR1D1 mediates abnormal proliferation and migration of PASMCs in⁃duced by hypoxia

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)

机构地区：[1]西部战区总医院烧伤科,四川成都610083 [2]西部战区总医院心血管内科,四川成都610083 [3]西南医科大学临床医学院,四川泸州646000 [4]西部战区总医院心脏外科,四川成都610083

出　　处：《中国病理生理杂志》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)。

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.

关 键 词：NR1D1蛋白 肺动脉平滑肌细胞 细胞增殖 细胞迁移 

分 类 号：R363[医药卫生—病理学] R543.2[医药卫生—基础医学] R364.4