双特异性磷酸酶9对低氧性肺动脉高压小鼠心肺损伤的改善作用实验研究

Improvement effects of dual specificity phosphatase 9 on cardiopulmonary injuryin mice with hypoxic pulmonary hypertension

目的:探讨双特异性磷酸酶9(DUSP9)对低氧性肺动脉高压(HPH)诱导的小鼠心肺损伤中的改善作用。方法:将C57BL/6小鼠60只随机分为对照组(NC组)、HPH组、HPH+空病毒组(HPH+vector组)和HPH+过表达DUSP9组(HPH+DUSP9组),每组15只。采用低压低氧人工舱建立HPH小鼠模型。于模型建立3周前,按分组要求尾静脉注射AAV9腺相关病毒及对照病毒。检测小鼠肺组织DUSP9蛋白表达、右心功能、心肌纤维化、肺血管重塑、血管活性物质水平、肺泡灌洗液炎症因子表达以及相对肺重量。结果:与NC组比较,HPH组和HPH+vector组DUSP9蛋白表达降低,右心室收缩压(RVSP)、右心室前壁厚度(RVAW)、右心室肥厚指数(RVHI)增加,右心室内径(RVID)降低,右心室胶原容积分数(CVF)增加,一氧化氮(NO)、总一氧化氮合酶(t-NOS)和诱导型一氧化氮合酶(iNOS)水平减少,内皮素-1(ET-1)水平增加,远端肺动脉壁厚度比(WT%)和肺动脉壁面积比(WA%)增加,肺泡灌洗液白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α及相对肺重量增加(均P<0.05)。与HPH组和HPH+vector组比较,HPH+DUSP9组DUSP蛋白表达增加,RVSP、RVAW、RVHI降低,RVID增加,右心室CVF降低,NO、t-NOS和iNOS水平增加,ET-1水平降低,WT%和WA%降低,肺泡灌洗液IL-1β、IL-6、TNF-α及相对肺重量降低(均P<0.05)。结论:过表达DUSP9可降低HPH小鼠肺动脉压力,改善小鼠右心功能、心肌纤维化、肺血管重塑、血管活性物质的动态平衡和肺内炎症水平,DUSP9可能是HPH小鼠心肺损伤的重要保护性分子。

Objective:To explore the improvement effects of dual specificity phosphatase 9(DUSP9)on cardiopulmonary injury in hypoxic pulmonary hypertension(HPH)mice.Methods:Adult male C57BL/6 mice were randomly divided into control group(NC group),HPH group,HPH+vector group and HPH+DUSP9 group.Artificial low oxygen tank is adopted to establish the HPH model in mice.AAV9 adeno-associated virus and control virus were injected into the tail vein 3 weeks before the establishment of the model according to the grouping requirements.DUSP9 protein expression,right ventricular cardiac function,myocardial fibrosis,pulmonary artery remodeling,vasoactive substance level,levels of inflammatory factors in alveolar lavage fluid,and relative lung weight were evaluated.Results:Compared with the NC group,the expression of DUSP9 protein was decreased,RVSP,RVAW,RVHI were increased,RVID was decreased,CVF was increased,NO,t-NOS and iNOS levels were decreased,ET-1 level was increased,WT%and WA%were increased,and levels of IL-1β,IL-6,TNF-αin bronchoalveolar lavage fluid and relative lung weight were increased in the HPH and HPH+vector groups(all P<0.05).Compared with HPH group and HPH+vector group,the expression of DUSP9 protein was increased,RVSP,RVAW,RVHI were decreased,RVID was increased,CVF was decreased,NO,t-NOS and iNOS levels were increased,ET-1 level was decreased,WT%and WA%were decreased,and the levels of IL-1β,IL-6,TNF-αin bronchoalveolar lavage fluid and relative lung weight were decreased in the HPH+DUSP9 group(all P<0.05).Conclusion:Overexpression of DUSP9 can reduce pulmonary artery pressure,improve right heart function,myocardial fibrosis,pulmonary vascular remodeling,dynamic balance of vasoactive substances and lung inflammation in HPH mice.DUSP9 may be an important protective molecule against cardiopulmonary injury in HPH mice.