VitD通过影响VDR/Sirt-1相互作用拮抗低氧诱导的肺动脉内皮细胞凋亡  

Antagonistic effect of VitD on hypoxia-induced apoptosis in human pulmonary artery endothelial cells by regulating VDR/Sirt-1 expression

作　　者：王蕾[1] 黄婧 Wang Lei;Huang Jing(Department of Pulmonary and Critical Care Medicine,the Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an710004,China;Department of Rheumatism and Immunology,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an710061,China)

机构地区：[1]西安交通大学第二附属医院呼吸与危重症医学科,西安710004 [2]西安交通大学第一附属医院风湿免疫科,西安710061

出　　处：《国际呼吸杂志》2022年第23期1823-1828,共6页International Journal of Respiration

基　　金：国家自然科学基金(82270055);中国博士后科学基金会(2021M702610);中央高校基本科研业务费(xzy012020060)。

摘　　要：目的探讨肺动脉高压患者维生素D/维生素D受体(VitD/VDR)表达情况及VitD对于低氧诱导的肺动脉内皮细胞(HPAECs)凋亡的作用。方法本研究为病例对照研究,采用非随机抽样的方法选取2016年1月至2020年12月西安交通大学第一附属医院结缔组织病合并肺动脉高压(CTD-PH)患者164例,收集患者血清,化学发光法检测患者血清25(OH)D水平;随机抽取20例CTD-PH患者提取外周血单个核细胞(PBMC),Real time PCR检测VDR mRNA表达。体外培养HPAECs,分为常氧组、常氧+VitD组、低氧组、低氧+VitD组,流式细胞仪检测细胞凋亡,蛋白免疫印迹法(Western blot)检测VDR、Sirt-1胞浆及胞核蛋白的表达。结果164例CTD-PH组患者25(OH)D浓度为(33.473±10.428)nmol/L,较国际标准50 nmol/L低,提示CTD-PH患者存在VitD缺乏。20例CTD-PH患者PBMCs VDR mRNA表达量为(1.635±0.508),高于健康对照组PBMCs VDR mRNA水平(1.019±0.198),差异有统计学意义(t=3.20,P<0.001)。体外细胞培养结果显示:低氧组细胞凋亡率高于常氧组(10.127±0.179)%比(3.503±0.181)%;常氧+VitD组凋亡率(1.280±0.181)%低于常氧组;低氧+VitD组凋亡率(5.993±0.181)%低于低氧组。检测VDR、Sirt-1蛋白表达结果显示:低氧组VDR、Sirt-1胞核蛋白较常氧组增加(P<0.05);低氧+VitD组与低氧组相比VDR胞浆蛋白、胞核蛋白均增加(P<0.05),Sirt-1胞浆蛋白、胞核蛋白均增加(P<0.05)。结论CTD-PH患者存在VitD缺乏,但VDR基因表达适应性增加,VitD可通过调控VDR/Sirt-1的表达,缓解低氧诱导的HPAECs凋亡,发挥保护作用。Objective To explore the expression of vitamin D/Vitamin D receptor(VitD/VDR)in patients with pulmonary hypertension(PH)and its effect on hypoxia-induced apoptosis in human pulmonary artery endothelial cells(HPAECs).Methods This study is a case-control,non-randomized study.A chemiluminescence method was conducted to detect serum 25-hydroxyvitamin D(25(OH)D)which were collected from 164 patients with connective tissue disease-pulmonary hypertension(CTD-PH)admitted to The First Affiliated Hospital of Xi′an Jiaotong University(January 2016 to December 2020).The peripheral blood mononuclear cells(PBMCs)were randomly extracted from 20 CTD-PH patients,and the real-time polymerase chain reaction(PCR)was used to detect the expression of VDR mRNA.The in-vitro cultured HPAECs were assigned to normoxic group,normoxic+VitD group,hypoxia group and hypoxia+VitD group.Flow cytometry were used for apoptosis detection,Western blot was performed to assess the nuclear protein and cytoplasmic protein of VDR,Sirtuin 1(Sirt1).Results The 25(OH)D level of 164 CTD-PH patients(33.473+10.428 nmo/L)was significantly lower than the international standard(50 nmo/L),indicating VitD deficiency in CTD-PH patients.The expression of VDR mRNA in PBMCs of randomized CTD-PH patients was significantly higher than that in healthy controls(1.635±0.508 vs 1.019±0.198).The result of in-vitro cultured HPAECs showed:the cell apoptosis rate of hypoxia group was higher than that of normoxic group(10.127±0.179 vs 3.503±0.181)%.The apoptosis rate of normoxic+VitD group(1.280±0.181)%was lower than normoxic group.The apoptosis rate of hypoxia+VitD group(5.993±0.181)%was lower than hypoxia group.The nuclear protein expression of VDR and Sirt-1 in hypoxia group was higher than normoxic group(P<0.05).Compared with the hypoxia group,the hypoxia+VitD group had elevated cytoplasm and nuclear protein of VDR and Sirt-1(P<0.05).Conclusions CTD-PH patients have VitD deficiency,whereas the expression of VDR gene is adaptively increased.VitD can alleviate hypoxia-in

关 键 词：肺动脉高压 细胞凋亡 维生素D 低氧 

分 类 号：R544.1[医药卫生—心血管疾病]