Bax抑制因子1可抑制体外培养的大鼠血管平滑肌细胞的钙化  

Bax inhibitor-1 inhibits calcification of vascular smooth muscle cells in vitro

作　　者：陈韦任 杨霞 周玉杰[1] 马茜[1] 吴雪萍 沙媛 钱赓 CHEN Weiren;YANG Xia;ZHOU Yujie;MA Qian;WU Xueping;SHA Yuan;QIAN Geng(Department of Cardiology,Beijing Anzhen Hospital of Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Disease,Beijing Key Laboratory of Precision Medicine of Coronary Atherosclerotic Disease,Beijing 100029,China;Department of Cardiology,Second Medical Center,First Medical Center,General Hospital of PLA,Beijing 100853,China;Department of Cardiology,First Medical Center,General Hospital of PLA,Beijing 100853,China)

机构地区：[1]首都医科大学附属北京安贞医院心内12病房,北京市心肺血管疾病研究所,冠心病精准治疗北京市重点实验室,北京100029 [2]中国人民解放军总医院第二医学中心心血管内科,北京100853 [3]中国人民解放军总医院第一医学中心心血管内科,北京100853

出　　处：《南方医科大学学报》2021年第8期1177-1182,共6页Journal of Southern Medical University

基　　金：国家重点研发计划“精准医学研究”重点专项(2017YFC0908800);北京市医院管理局“使命”计划专项经费资助(SML20180601);首都卫生发展科研专项(首发2020-2-2063);北京市教育委员会科技计划(KM200910025012);北京市自然科学基金(7202041);北京市博士后资助项目(202011)。

摘　　要：目的探讨Bax抑制因子1(BI-1)对血管平滑肌细胞钙化的具体作用机制。方法使用β磷酸甘油和氯化钙诱导大鼠主动脉血管平滑肌细胞建立钙化模型。将模型分为对照组(使用常规培养基)、BI-1过表达组(过表达BI-1蛋白后使用常规培养基)、钙化组(使用钙化培养基)、钙化+BI-1过表达组(过表达BI-1蛋白后使用钙化培养基)4组。运用茜素红S染色测定血管平滑肌细胞钙沉积,酶联免疫吸附法测定细胞碱性磷酸酶活性和钙含量,Western blot法测定Runt相关转录因子2、骨形态发生蛋白2和半胱氨酸天冬氨酸蛋白酶3表达水平。结果(1)随着钙化诱导时间的延长,BI-1蛋白表达明显下降,结果具有明显差异性(P=0.001);(2)使用质粒转染方法过表达BI-1蛋白后,细胞钙含量、碱性磷酸酶活性明显降低(P=0.0006),Runt相关转录因子2和骨形态发生蛋白2表达明显下降(P=0.0001),血管钙化减轻;(3)钙化诱导后血管平滑肌细胞凋亡率增加,而过表达BI-1后细胞凋亡率明显减少(P=0.01),半胱氨酸天冬氨酸蛋白酶3水平下降(P=0.0003)。结论BI-1抑制细胞凋亡、钙磷沉积和细胞骨型分化起到减轻血管钙化作用。Objective To investigate the effect of Bax inhibitor-1(BI-1)on calcification of vascular smooth muscle cells(VSMCs).Methods VSMCswere isolated from the thoracic aorta of SD rats.VSMCs or BI-1-overexpressing VSMCs(transfected with a BI-1-overexpressing plasmid)were cultured in normal medium or calcified medium containingβ-glycerophosphate and calcium chloride,and the cell calcification was examined with Alizarin red staining.Enzyme-linked immunosorbent assay was used to determine the intracellular calcium content and alkaline phosphatase activity.The expression levels of Runt-related transcription factor 2(RUNX2),bone morphogenetic protein 2(BMP-2)and caspase-3 were detected with Western blotting.Results After 14 days of culture in the calcified medium,the VSMCs showed significantly reduced expression of BI-1 protein(P=0.001).BI-1 overexpression in the VSMCs caused a significant reduction of calcium level and alkaline phosphatase activities(P=0.0006)and lowered the expression levels of RUNX2 and BMP-2(P=0.0001)in the cells.The VSMCs with induced calcification exhibited a significantly increased apoptosis rate,but BI-1 overexpression obviously inhibited VSMC apoptosis in the calcified medium(P=0.0003).Conclusion BI-1 may attenuate vascular calcification by inhibiting calcium deposition,osteogenic differentiation and apoptosis.

关 键 词：Bax inhibitor-1 血管钙化 细胞成骨型分化 钙沉积 细胞凋亡 

分 类 号：R543[医药卫生—心血管疾病]