激活ALDH2通过上调SIRT1/PGC-1α信号通路减轻小鼠缺氧性肺动脉高压  

Activation of ALDH2 alleviates hypoxic pulmonary hypertension in mice by upregulating the SIRT1/PGC-1αsignaling pathway

作　　者：王磊 卞芬兰 马飞扬 方舒 凌梓涵 刘梦然 孙红燕 付程文 倪诗垚 赵晓阳 冯心茹 孙正宇 卢国庆 康品方 吴士礼 WANG Lei;BIAN Fenlan;MA Feiyang;FANG Shu;LING Zihan;LIU Mengran;SUN Hongyan;FU Chengwen;NI Shiyao;ZHAO Xiaoyang;FENG Xinru;SUN Zhengyu;LU Guoqing;KANG Pinfang;WU Shili(Department of Cardiology,First Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China;Key Laboratory of Cardio-Cerebrovascular Foundation and Clinical Sciences,Bengbu Medical University,Bengbu 233000,China;Clinical College of Medicine,Bengbu Medical University,Bengbu 233000,China;Department of Anesthesiology,First Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China;Department of Otolaryngology,First Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China)

机构地区：[1]蚌埠医科大学第一附属医院心血管科,安徽蚌埠233000 [2]蚌埠医科大学心脑血管基础与临床重点实验室,安徽蚌埠233000 [3]蚌埠医科大学临床医学院,安徽蚌埠233000 [4]蚌埠医科大学第一附属医院麻醉科,安徽蚌埠233000 [5]蚌埠医科大学第一附属医院耳鼻喉科,安徽蚌埠233000

出　　处：《南方医科大学学报》2024年第10期1955-1964,共10页Journal of Southern Medical University

基　　金：国家自然科学基金(81970313);安徽省自然科学基金(2208085MH192);安徽省高校协同创新项目(GXXT-2020-019);安徽省优秀青年基金(2022AH030141);安徽省优秀创新团队(2022AH010083);蚌埠医学院研究生科研创新计划项目(Byycx23099);安徽省临床转化课题青年项目(202304295107020034)。

摘　　要：目的探讨激活线粒体乙醛脱氢酶2(ALDH2)是否可以通过SIRT1/PGC-1α信号通路减轻缺氧性肺动脉高压。方法在体水平:选取8周龄C57 BL/6小鼠40只,随机分成Control组、Hypoxia组、Hypoxia+Alda-1灌胃组及ALDH2特异性敲除组(Hypoxia+ALDH2^(-/-)),10只/组,Hypoxia组小鼠暴露于缺氧条件下(10%O_(2),90%N_(2)),维持每日12 h/12 h的暗/光循环4周,Hypoxia+Alda-1组同时予以Alda-1腹腔注射处理4周,Control组予以常氧环境并给与同等量的溶剂(DMSO+PBS)干预4周。利用超声心动图、右心室导管实验评估右心功能及压力,以右心室压代替肺动脉压;HE评估肺血管重构及右心室损伤情况,免疫荧光检测α-SMA表达,评估肺远端小动脉肌化情况,WGA染色检测右心室横截面积,评估心肌细胞肥大程度,测量右心肥厚指数。检测ALDH2、SIRT1、PGC-1α、P16^(INK4A)、P21^(CIP1)蛋白表达。体外水平:分别设置Control组、Hypoxia组上、Hypoxia+Alda-1组上、Hypoxia+Alda-1+EX527组。利用β半乳糖染色评估各分组小鼠肺动脉平滑肌细胞衰老情况,Western blotting评估各分组蛋白表达情况。结果与Control组相比,Hypoxia组右心室收缩压(RVSP)增高、右室游离壁厚度(RVFWT)增厚、P16^(INK4A)、P21^(CIP1)表达增加(P<0.05)。与Hypoxia组相比,Hypoxia+Alda-1组RVSP降低、RVFWT变薄、P16^(INK4A)、P21^(CIP1)表达降低(P<0.05)。与Hypoxia组相比,Hypoxia+ALDH2^(-/-)组RVSP增高、RVFWT增厚、P16^(INK4A)、P21^(CIP1)蛋白表达增加(P<0.01)。与Control组相比,Hypoxia组细胞衰老比例增加、P16^(INK4A)、P21^(CIP1)表达增加(P<0.01)。与Hypoxia组相比,Hypoxia+Alda-1组细胞衰老比例降低(P<0.01),P16^(INK4A)、P21^(CIP1)(P<0.05)表达降低。与Hypoxia+Alda-1组相比,Hypoxia+Alda-1+EX527组细胞衰老比例增加(P<0.01),P16^(INK4A)、P21^(CIP1)表达增高(P<0.05)。结论ALDH2通过调控SIRT1/PGC-1α信号通路,减轻小鼠肺动脉平滑肌细胞衰老,从而减轻缺氧型肺动脉高压。Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1αsignaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2^(-/-))mice,were exposed to hypoxia(10%O_(2),90%N_(2))with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardialα-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16^(INK4A) and P21^(CIP1) were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated usingβ-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16^(INK4A) and P21^(CIP1),which were effectively lowered by treatment with Alda-1 but further increased in ALDH2^(-/-)mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16^(INK4A) and P21^(CIP1),which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxiainduced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1αsignaling pathway to alleviate pulmonary hypertension in mice.

关 键 词：线粒体乙醛脱氢酶2 SIRT1 PGC-1Α 平滑肌细胞衰老 肺动脉高压 

分 类 号：R544.1[医药卫生—心血管疾病]