PCK1对小鼠血管平滑肌细胞增殖和迁移的作用及其机制  

Effect of PCK1 on proliferation and migration of mouse vascular smooth muscle cells and its underlying mechanism

作　　者：张黎 王嘉[3] 方世正 张钟健 杨曦 王武帅 孙雄山 杨大春 ZHANG Li;WANG Jia;FANG Shizheng;ZHANG Zhongjian;YANG Xi;WANG Wushuai;SUN Xiongshan;YANG Dachun(Department of Cardiovascular Medicine,The Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;Department of Cardiovascular Medicine,The General Hospital of Western Theater Command,Chengdu 610083,China;College of Medicine,Southwest Jiaotong University,Chengdu 610031,China)

机构地区：[1]西南医科大学附属医院心血管内科,四川泸州646000 [2]西部战区总医院心内科,四川成都610083 [3]西南交通大学医学院,四川成都610031

出　　处：《中国病理生理杂志》2024年第6期971-979,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82100419);西部战区总医院“星火”创新人才项目。

摘　　要：目的:探讨磷酸烯醇式丙酮酸羧激酶1(PCK1)在小鼠血管平滑肌细胞(VSMCs)增殖及迁移中的作用及机制。方法:用30μg/L血小板源性生长因子BB(PDGF-BB)诱导小鼠VSMCs增殖和迁移,将小鼠VSMCs分为溶剂对照(vehicle)组和PDGF-BB组,采用Western blot和免疫荧光染色检测PCK1表达水平的变化。使用小鼠Pck1 siRNA(siPck1)转染小鼠VSMCs沉默PCK1表达,将VSMCs分为vehicle组、siPck1+vehicle组、PDGF-BB组和siPck1+PDGF-BB组,采用免疫荧光染色检测细胞增殖能力,CCK-8法检测细胞活力,划痕实验检测细胞迁移能力,透射电镜观察细胞线粒体动力学变化。发动蛋白相关蛋白1(Drp1)基因过表达慢病毒(lenti-Drp1)转染VSMCs使DRP1过表达,将小鼠VSMCs分为PDGF-BB组、siPck1+PDGF-BB组、lenti-Drp1+PDGF-BB组和lenti-Drp1+siPck1+PDGF-BB组,再次检测上述指标。结果:PDGF-BB使VSMCs中PCK1和DRP1表达增加,细胞活力升高,Ki-67阳性细胞率增加,划痕愈合率升高,线粒体分裂增加;沉默PCK1表达后以上过程均受到抑制。过表达DRP1后,沉默PCK1表达对VSMCs细胞活力、Ki-67阳性细胞率、划痕愈合率和线粒体分裂的抑制作用明显削弱。结论:PCK1通过调控DRP1表达促进小鼠VSMCs线粒体分裂、细胞增殖和迁移。AIM:To investigate the role of phosphoenolpyruvate carboxykinase 1(PCK1)in the proliferation and migration of mouse vascular smooth muscle cells(VSMCs)and the underlying mechanism.METHODS:The proliferation and migration of mouse VSMCs were induced by platelet-derived growth factor(PDGF)-BB.The cells were divided into a vehicle group and a PDGF-BB group.The expression of PCK1 was detected by Western blot and immunofluorescence staining.The mouse Pck1 siRNA(siPck1)were transfected into mouse VSMCs to silence PCK1.The cells were divided into the vehicle,siPck1+vehicle,PDGF-BB and siPck1+PDGF-BB groups.The protein level of PCK1 was detected by Western blot.The proliferation was explored by Ki-67 immunofluorescence staining and the viability was detected by CCK-8 assay.The migration was determined by a scratch test.Mitochondrial dynamics were observed via transmission electron microscopy.A lentivirus carrying dynamin-related protein 1(Drp1)gene(lenti-Drp1)was transfected into VSMCs to induce them to overexpress DRP1.The cells were divided into the PDGF-BB,siPck1+PDGF-BB,lenti-Drp1+PDGF-BB and lenti-Drp1+siPck1+PDGF-BB groups.Proliferation,migration and mitochondrial dynamics were measured as described above.RESULTS:PDGF-BB increased the protein expression of PCK1 and DRP1,cell viability,the percentage of Ki-67-positive cells,the wound healing rate and mitochondrial division in VSMCs.These effects were suppressed when PCK1 protein expression was silenced.After DRP1 was overexpressed,the inhibitory effects of PCK1 silencing on cell viability,the percentage of Ki-67-positive cells,the wound healing rate and mitochondrial division were significantly reversed.CONCLUSION:PCK1 promotes the mitochondrial division,proliferation and migration of VSMCs in mice by upregulating the expression of DRP1.

关 键 词：磷酸烯醇式丙酮酸羧激酶1 血管平滑肌细胞 细胞增殖 细胞迁移 线粒体动力学 

分 类 号：R543.5[医药卫生—心血管疾病] R363.2[医药卫生—内科学] R977.3[医药卫生—临床医学]