南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (8): 1177-1182.doi: 10.12122/j.issn.1673-4254.2021.08.08

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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

陈韦任，杨 霞，周玉杰，马 茜，吴雪萍，沙 媛，钱 赓   

  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；中国人民解放军总医院第二医学中心心血管内科，第一医学中心心血管内科，北京 100853
  • 出版日期:2021-08-20 发布日期:2021-09-07

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

CHEN Weiren, YANG Xia, ZHOU Yujie, MA Qian, WU Xueping, SHA Yuan, QIAN Geng   

  1. 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, Department of Cardiology, First Medical Center, General Hospital of PLA, Beijing 100853, China
  • Online:2021-08-20 Published:2021-09-07

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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。

关键词: Bax inhibitor-1；血管钙化；细胞成骨型分化；钙沉积；细胞凋亡

Abstract: Objective To investigate the effect of Bax inhibitor-1 (BI-1) on calcification of vascular smooth muscle cells (VSMCs). Methods VSMCs were 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 containing β- 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.

Key words: Bax inhibitor-1; vascular calcification; osteogenic differentiation; calcium deposition; apoptosis