南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (3): 330-337.doi: 10.12122/j.issn.1673-4254.2022.03.03

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Bax抑制因子1通过促进视神经萎缩蛋白1表达抑制小鼠动脉血管钙化

陈韦任，杜 辉，钱 赓，周玉杰，陈韵岱，马 茜，吴雪萍，沙 媛   

  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，首都医科大学冠心病临床诊疗与研究中心，北京 100029；中国人民解放军总医院第二医学中心心血管内科，国家老年疾病临床医学研究中心，北京 100853；中国人民解放军总医院第一医学中心心血管内科，北京 100853
  • 出版日期:2022-03-20 发布日期:2022-04-11

Bax inhibitor 1 inhibits vascular calcification in mice by activating optic atrophy 1 expression

CHEN Weiren, DU Hui, QIAN Geng, ZHOU Yujie, CHEN Yundai, MA Qian, WU Xueping, SHA Yuan   

  1. Department of Cardiology, Beijing Anzhen Hospital of Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Disease; Department of Cardiology, Second Medical Center and National Clinical Research Center for Geriatric Diseases, Chinese PLA General Hospital; Department of Cardiology, First Medical Center, Chinese PLA General Hospital
  • Online:2022-03-20 Published:2022-04-11

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摘要： 目的 探讨Bax抑制因子1（BI-1）和视神经萎缩蛋白1（OPA1）蛋白对血管钙化的影响。方法 ApoE-/-糖尿病小鼠高脂喂养12周后予以腹腔注射Nε（- 1-羧甲基）-L-赖氨酸16周建立血管钙化模型，实验将小鼠分为4组，6只/组：对照组（ApoE-/-小鼠普通饲料喂养）、钙化组（ApoE-/-小鼠建立钙化模型）、钙化+BI-1TG组（血管特异性过表达BI-1的ApoE-/-小鼠建立钙化模型）和钙化＋BI-1TG＋OPA1-/-组（血管特异性过表达BI-1和基因敲除OPA1的ApoE-/-小鼠建立钙化模型）。另β磷酸甘油诱导血管平滑肌细胞建立细胞钙化模型。使用von Kossa染色检测血管钙化程度，使用ELISA检测主动脉钙含量，使用免疫组织化学方法检测Runt相关转录因子2和骨形态发生蛋白2表达、使用TUNEL检测细胞凋亡率，使用Western blot检测BI-1、OPA1、Runt相关转录因子2、骨形态发生蛋白2和活化的半胱氨酸天冬氨酸蛋白酶3水平的表达。结果 血管钙化后，BI-1和OPA1蛋白表达均下降（P=0.0044），钙沉积、Runt相关转录因子2、骨形态发生蛋白2及活化的半胱氨酸天冬氨酸蛋白酶3表达增加（P=0.0041）。过表达BI-1促进OPA1蛋白表达，钙沉积、Runt相关转录因子2、骨形态发生蛋白2及活化的半胱氨酸天冬氨酸蛋白酶3表达均减少（P=0.0006）。基因敲除OPA1蛋白后，钙沉积、Runt相关转录因子2、骨形态发生蛋白2及活化的半胱氨酸天冬氨酸蛋白酶3又明显增多（P=0.0007）。结论 BI-1可能通过促进OPA1表达，减轻钙沉积、细胞骨型分化和细胞凋亡，继而抑制血管钙化。

关键词: Bax抑制因子1；视神经萎缩蛋白1；血管钙化；细胞骨型分化；细胞凋亡

Abstract: Objective To investigate the effects of Bax inhibitor 1 (BI- 1) and optic atrophy protein 1 (OPA1) on vascular calcification (VC). Methods Mouse models of VC were established in ApoE-deficient (ApoE-/-) diabetic mice by high-fat diet feeding for 12 weeks followed by intraperitoneal injections with Nε-carboxymethyl-lysine for 16 weeks. ApoE-/- mice (control group), ApoE-/- diabetic mice (VC group), ApoE-/- diabetic mice with BI-1 overexpression (VC + BI-1TG group), and ApoE-/-diabetic mice with BI-1 overexpression and OPA1 knockout (VC+BI-1TG+OPA1-/- group) were obtained for examination of the degree of aortic calcification using von Kossa staining. The changes in calcium content in the aorta were analyzed using ELISA. The expressions of Runt-related transcription factor 2 (RUNX2) and bone morphogenetic protein 2 (BMP-2) were detected using immunohistochemistry, and the expression of cleaved caspase-3 was determined using Western blotting. Cultured mouse aortic smooth muscle cells were treated with 10 mmol/L β-glycerophosphate for 14 days to induce calcification, and the changes in BI-1 and OPA1 protein expressions were examined using Western blotting and cell apoptosis was detected using TUNEL staining. Results ApoE-/- mice with VC showed significantly decreased expressions of BI-1 and OPA1 proteins in the aorta (P=0.0044) with obviously increased calcium deposition and expressions of RUNX2, BMP-2 and cleaved caspase-3 (P=0.0041). Overexpression of BI-1 significantly promoted OPA1 protein expression and reduced calcium deposition and expressions of RUNX2, BMP-2 and cleaved caspase-3 (P=0.0006). OPA1 knockdown significantly increased calcium deposition and expressions of RUNX2, BMP-2 and cleaved caspase-3 in the aorta (P=0.0007). Conclusion BI-1 inhibits VC possibly by promoting the expression of OPA1, reducing calcium deposition and inhibiting osteogenic differentiation and apoptosis of the vascular smooth muscle cells.

Key words: Bax inhibitor 1; optic atrophy protein 1; vascular calcification; osteogenic differentiation; apoptosis