缺氧诱导BMPR2突变大鼠左心室损伤  

Hypoxia Induced Left Ventricular Injury in BMPR2 Mutant Rats

作　　者：张学佳 李天骐 李平[1] 马铭婕 吕婷婷 潘慧[1] 纪爽 叶梦婷 王晓建[1,4,5] ZHANG Xuejia;LI Tianqi;LI Ping;MA Mingjie;LYU Tingting;PAN Hui;JI Shuang;YE Mengting;WANG Xiaojian(Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,Key Laboratory of Pulmonary Vascular Medicine,National Center for Cardiovascular Diseases,Chinese Academy of Medical Sciences&Peking Union Medical College,Bejing 100037,China;The First Hospital of China Medical University,Shenyang 110000,Liaoning,China;Quzhou Kecheng District People's Hospital,Quzhou 324000,Zhejiang,China;Fuwai Yunnan Hospital,Chinese Academy of Medical Sciences,Kunming 650102,Yunnan,China;National Health Commission Key Laboratory of Cardiovascular Regenerative Medicine,Fuwai Central China Cardiovascular Hospital,Zhengzhou 451464,Henan,China)

机构地区：[1]中国医学科学院北京协和医学院肺血管医学重点实验室国家心血管病中心心血管疾病国家重点实验室阜外医院,北京100037 [2]中国医科大学附属第一医院,沈阳110000 [3]衢州市柯城区人民医院,浙江衢州324000 [4]云南省阜外心血管病医院,昆明650102 [5]国家卫生健康委员会心血管疾病再生医学重点实验室阜外华中心血管病医院,郑州451464

出　　处：《中国分子心脏病学杂志》2024年第3期6128-6135,共8页Molecular Cardiology of China

基　　金：国家自然科学基金面上项目(82370064,82170408);北京市自然科学基金面上项目(7222142);中国医学科学院阜外医院高水平医院临床科研攻关项目(2023-GSP-GG-24)。

摘　　要：目的研究骨形态发生蛋白2型受体(BMPR2)基因突变对左心室结构与功能的影响。方法利用CRISPR-Cas9技术构建Bmpr2^(+/R491W)突变型大鼠,在基线期以及单纯低氧[吸入氧浓度(FiO_(2))=10%]刺激3周后通过超声、左右心导管以及病理实验检测大鼠左心室、右心室、肺血管表型;通过定量聚合酶链反应(quantitative polymerase chain reaction,QPCR)检测心力衰竭分子标志物,通过免疫印迹检测BMPR2通路活性。结果基线情况下,Bmpr2^(+/R491W)突变型大鼠无明显表型异常。经过3周低氧刺激,Bmpr2^(+/R491W)突变型大鼠与野生型大鼠均发生了肺动脉高压,肺动脉压力与右心室重构程度相似,差异无统计学意义。低氧刺激未影响野生型大鼠的左心室结构与功能,但诱导了Bmpr2^(+/R491W)突变型大鼠左心室明显重构;Bmpr2^(+/R491W)突变型大鼠在缺氧后左室射血分数显著下降(78.7%±6.7%vs.66.4%±10.9%),P<0.05),左心室短轴缩短率下降(50.0%±7.3%vs.(38.5%±8.6%,P<0.05),收缩期左心室后壁厚度减小(3.3%±0.4%vs.2.4%±0.6%,P<0.01)。QPCR检测显示心房利尿钠肽、脑利尿钠肽等心力衰竭分子标志物表达显著上调,免疫印迹检测结果表明左心组织BMPR2表达降低,为对照组的50%,ID-1下降至对照组的60%,而p-SMAD2/3比例上调0.6倍。结论缺氧刺激诱导Bmpr2^(+/R491W)大鼠在低氧刺激下产生肺动脉高压表型,并且发生左心衰竭及功能障碍。Objective To study the effect of the bone morphogenetic protein receptor type 2(BMPR2)gene mutation on the structure and function of the left ventricle.Methods Used CRISPR-Cas9 technology to construct Bmpr2^(+/R491W)mutant rats.Evaluated the phenotype of the left and right ventricles and pulmonary vessels at baseline and after 3 weeks of hypoxia(FiO_(2)=10%)stimulation using echocardiography,cardiac catheterization,and pathological experiments.Heart failure molecular markers were assessed by quantitative polymerase chain reaction(QPCR),and BMPR2 pathway activity was measured by Western blot.Results At baseline,the Bmpr2^(+/R491W)mutant rats exhibited no significant phenotypic abnormalities.After 3 weeks of hypoxic stimulation,both Bmpr2^(+/R491W)mutant rats and wild-type rats developed pulmonary arterial hypertension,with similar levels of pulmonary artery pressure and right ventricular reconstruction,with no statistical difference.Hypoxic stimulation did not affect the left ventricular structure and function in wild-type rats but induced significant left ventricular remodeling in Bmpr2^(+/R491W)mutant rats.The Bmpr2^(+/R491W)mutant rats showed a significant decrease in left ventricular ejection fraction(78.7%±6.7%vs.66.4%±10.9%,P<0.05),left ventricular fractional shortening(50.0%±7.3%vs.38.5%±8.6%,P<0.05),,and diastolic left ventricular posterior wall thickness(3.3%±0.4%vs.2.4%±0.6%,P<0.01)post hypoxia.QPCR showed significant upregulation of heart failure molecular markers such as atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),etc.,and Western blot results indicated that left heart tissue BMPR2expression was reduced to 50%of the control group,ID-1 was reduced to 60%of the control group,and the ratio of phosphorylated SMAD2/3 increased by 0.6 times.Conclusion Hypoxic stimulation induced a pulmonary arterial hypertension phenotype and left heart failure and dysfunction inBmpr2+/R491Wrats under hypoxic conditions.

关 键 词：骨形态发生蛋白2型受体 肺动脉高压 基因突变 左心室重构 

分 类 号：R54[医药卫生—心血管疾病]