特异性敲减平滑肌细胞XBP-1对低氧性肺动脉高压小鼠心肺损伤的抑制作用  

Inhibitory effect of specific knockdown of XBP-1 in smooth muscle cells on cardiopulmonary injury in hypoxia-induced pulmonary hypertension mice

作　　者：吴宾 杨自更 李彩玲 梁羽薇 宁海虹 王嘉玮 黄海 WU Bin;YANG Zigeng;LI Cailing;LIANG Yuwei;NING Haihong;WANG Jiawei;HUANG Hai(Department of Nuclear Medicine,General Hospital of Xinjiang Military Command of the Chinese People's Liberation Army,Urumqi 830000,China;不详)

机构地区：[1]中国人民解放军新疆军区总医院核医学科,乌鲁木齐830000 [2]中国人民解放军新疆军区总医院卫勤训练中心 [3]中国人民解放军新疆军区总医院肾内科

出　　处：《山东医药》2024年第26期47-52,共6页Shandong Medical Journal

基　　金：新疆维吾尔自治区自然科学基金面上项目(2022D01C644);新疆军区总医院喀喇昆仑人才基金项目(2022JC002)。

摘　　要：目的探讨特异性敲减平滑肌细胞X盒结合蛋白1(XBP-1)对低氧性肺动脉高压(HPH)小鼠心肺损伤的抑制作用。方法取8周龄雄性C57BL/6小鼠60只,随机分为对照组、HPH组、HPH+对照病毒(HPH+shCtrl)组、HPH+敲减XBP-1(HPH+shXBP-1)组。对照组置于常压常氧环境饲养,HPH组、HPH+shCtrl组、HPH+shXBP-1组置于低压低氧环境进行HPH造模;于造模前3周,HPH+shXBP-1组尾静脉注射平滑肌特异性XBP-1敲减的AAV9病毒,HPH+shCtrl组尾静脉注射对照病毒。将小鼠麻醉后,采用心导管检测右心室收缩压(RVSP);小动物超声仪检测右心室内径(RVID)、右心室前壁厚度(RVAW);取心脏,测算右心室肥厚指数(RVHI);取右心室组织,进行Masson染色,计算胶原容积分数(CVF);眶周取血,采用ELISA法检测血浆内皮素1(ET-1),比色法检测一氧化氮、总一氧化氮合酶(NOS)、诱导型一氧化氮合酶(iNOS);对左肺进行肺泡灌洗,收集肺泡灌洗液,采用ELISA法检测白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α);取右肺中叶及后叶肺组织进行HE染色,评估远端肺动脉壁厚度比(WT%)和肺动脉壁面积比(WA%);取右肺前叶肺组织,提取组织总蛋白,采用Western blotting法检测XBP-1蛋白表达。结果HPH组和HPH+shCtrl组RVSP较对照组增加,HPH+shXBP-1组RVSP较HPH组和HPH+shCtrl组降低(P均<0.05)。与对照组比较,HPH组和HPH+shCtrl组RVID降低,RVAW、RVHI增加;与HPH组和HPH+shCtrl组比较,HPH+shXBP-1组RVID增加,RVAW、RVHI降低(P均<0.05)。HPH组和HPH+shCtrl组CVF较对照组升高,HPH+shXBP-1组CVF较HPH组和HPH+shCtrl组降低(P均<0.05)。HPH组和HPH+shCtrl组血浆ET-1水平较对照组升高,NO、总NOS和iNOS水平较对照组降低(P均<0.05);HPH+shXBP-1组血浆ET-1水平较HPH组和HPH+shCtrl组降低,NO、总NOS和iNOS水平较HPH组和HPH+shCtrl组升高(P均<0.05)。HPH组和HPH+shCtrl组肺泡灌洗液IL-1β、IL-6、TNF-α水平较对照组升高,HPH+shXBP-1组IL-1β、IL-6、TNF-α水平较HPHObjective To investigate the preventive effect of specific knockdown of X box-binding protein 1(XBP-1)in smooth muscle cells on cardiopulmonary injury in hypoxia-induced pulmonary hypertension(HPH)mice.Methods Sixty 8-week-old male C57BL/6 mice were selected and randomly divided into control group(NC),HPH group,HPH+shCtrl group,and HPH+shXBP-1 group,respectively.Mice in the control group was placed in normal pressure and oxygen environment,and mice in the HPH group,HPH+shCtrl group,and HPH+shXBP-1 group were placed in low pressure and low oxygen environment for HPH modeling.At 3 weeks prior to model construction,mice in the HPH+shXBP-1 group were injected with smooth muscle specific XBP-1 knockdown adeno-associated virus serotype 9(AAV9)virus through the tail vein,and mice in the HPH+shCtrl group were injected with control virus.After anesthesia,right ventricular systolic pressure(RVSP)was measured by cardiac catheter.Right ventricular internal diameter(RVID)and thickness of right ventricle anterior wall(RVAW)were measured with small animal ultrasound instrument.The heart was isolated and right ventricular hypertrophy index(RVHI)was evaluated.Right ventricular myocardium was collected and masson was performed to calculate collagen volume fraction(CVF).The blood of mice was collected from periorbital area,and endothelin-1(ET-1)was detected by enzyme-linked immunosorbent assay(ELISA)and the levels of nitric oxide(NO),nitric oxide synthase(NOS)and induced nitric oxide synthase(iNOS)were detected by colorimetry.Alveolar lavage was performed on the left lung,and the bronchoalveolar lavage fluid(BALF)was collected.Interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)were detected by ELISA.HE staining was performed on the middle and posterior lobe of the right lung,and the distal pulmonary artery wall thickness ratio(WT%)and pulmonary artery wall area ratio(WA%)were evaluated.The total protein of the right anterior lobe lung was extracted,and the expression of XBP-1 protein was detected by Western

关 键 词：低氧性肺动脉高压 X盒结合蛋白1 右心室收缩压 心肌纤维化 炎症反应 内质网应激 

分 类 号：R543.2[医药卫生—心血管疾病]