Page 42 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第43卷第8期
·1076 · Journal of Nanjing Medical University（Natural Sciences） 2023年8月

·基础研究·

穹窿主体蛋白通过上调干扰素调节因子2抑制动脉内皮细胞凋亡

薛佳佳 ，王艺颖 ，胡成秀 ，孙崇秀 1*
1
1，2
1
南京医科大学心血管疾病转化医学协同创新中心，江苏 南京 211166；江苏卫生健康职业学院药学院基础化学教研室，江
1 2
苏 南京 211800

［摘 要］ 目的：探究穹窿主体蛋白（major vault protein，MVP）对动脉内皮细胞（endothelial cell，EC）增殖的作用，揭示MVP对
动脉EC发挥保护作用的潜在机制。方法：以人主动脉EC（human aortic EC，HAEC）为细胞模型，感染慢病毒以抑制或过表达
MVP。使用CCK⁃8实验和流式细胞术检测细胞增殖活性和死亡。使用凋亡抑制剂Z⁃VAD和坏死性凋亡抑制剂Nec⁃1确定细
胞死亡方式。以流式细胞术检测Annexin V结合阳性率和Caspase 3活性，以Western blot检测Caspase剪切体蛋白表达以评价
细胞凋亡。以荧光定量 PCR 和 Western blot 技术鉴定靶分子，并明确 MVP 与靶分子之间的调控关系。结果：敲降 MVP 抑制
HAEC增殖，促进HAEC死亡，过表达MVP结果则相反。Z⁃VAD逆转MVP敲降引起的死亡，而Nec⁃1无此作用。过表达MVP抑
制TNF⁃α诱导的HAEC凋亡，敲降MVP时作用相反。MVP通过上调干扰素调节因子2（interferon regulatory factor 2，IRF2）促进
凋亡抑制蛋白1（cellular inhibitor of apoptosis protein 1，cIAP1）的转录。敲降IRF2逆转MVP过表达引起的cIAP1表达增多和凋亡
抑制。结论：MVP通过上调IRF2蛋白促进cIAP1转录表达从而抑制TNF⁃α诱导的HAEC凋亡，发挥对EC的保护作用。
［关键词］ 穹窿主体蛋白；动脉内皮细胞；细胞增殖；细胞凋亡；凋亡抑制蛋白1
［中图分类号］ R329.25 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）08⁃1076⁃09
doi：10.7655/NYDXBNS20230806

Major vault protein inhibits apoptosis of aortic endothelial cells through upregulating
interferon regulatory factor 2

1 1，2 1 1*
XUE Jiajia ，WANG Yiying ，HU Chengxiu ，SUN Chongxiu
1 Key Laboratory of Targeted Intervention of Cardiovascular Disease，Collaborative Innovation Center for
Cardiovascular Disease Translational Medicine，Nanjing Medical University，Nanjing 211166；Department of
2
General Chemistry，School of Pharmacy，Jiangsu Health Vocational College，Nanjing 211800，China

［Abstract］ Objective：To investigate the effects and the underlying mechanism of major vault protein（MVP）on the proliferation of
arterial endothelial cells. Methods：Human aortic endothelial cell（HAEC）were infected with lentivirus to inhibit or overexpress MVP.
Cell proliferation and death were detected with CCK⁃8 assay and flow cytometry. Apoptosis inhibitor Z⁃VAD and necroptosis inhibitor
Nec⁃1 were used to distinguish the mode of cell death. Annexin V binding and Caspase 3 activity were detected by flow cytometry，and
cleaved Caspase was examined by Western blot. Real time PCR and Western blot were performed to investigate target molecules and
the regulatory relationship. Results：Knockdown of MVP inhibited the proliferation activity of HAEC and promoted the HAEC cell
death. Overexpression of MVP resulted in the opposite results. Treatment with Z ⁃ VAD reversed HAEC death caused by MVP
knockdown，while Nec⁃1 did not. Consistently，TNF⁃α⁃induced HAEC apoptosis was inhibited by MVP overexpression and exaggerated
by MVP knocked down. MVP promoted the transcriptional expression of cellular inhibitor of apoptosis proteins1（cIAP1）by up ⁃
regulating interferon regulatory factor 2（IRF2）protein expression. IRF2 knockdown reversed the increase in cIAP1 expression and the
decrease in apoptosis caused by MVP overexpression. Conclusion：MVP promoted cIAP1 transcription by up⁃regulating IRF2 protein
expression，thereby inhibiting TNF⁃α⁃induced apoptosis and promoting the proliferation of arterial EC.
［Key words］ major vault protein；aortic endothelial cells；cell proliferation；cell apoptosis；cIAP1
［J Nanjing Med Univ，2023，43（08）：1076⁃1084］

［基金项目］ 国家自然科学基金（81870355，82170465）
∗
通信作者（Corresponding author），E⁃mail：cxsun@njmu.edu.cn