rBMSCs/F92A-Cav1对PAH大鼠的治疗作用及其机制

Therapeutic effect and mechanism of r BMSCs/F92A-Cav1 on PAH rats

目的探讨过表达F92A-丙氨酸替代窖蛋白-1(Cav1)的大鼠骨髓间充质干细胞(r BMSCs/F92A-Cav1)对肺动脉高压(PAH)大鼠的治疗作用及其机制。方法予成年雄性Wistar大鼠腹腔注射1%野百合碱(MCT,60 mg/kg)建立PAH模型,建模后2周随机分为3组(10只/组):PAH组(仅给予MCT)、Cav1组(转导LV-Cav1的r BMSCs)、F92A-Cav1组(转导LV-F92A-Cav1的r BMSCs),同时设置正常组。基因修饰的r BMSCs(1×106/m L)于尾静脉移植入各组PAH大鼠,移植后3周,采用Image-Pro Plus 6 software评估肺动脉中膜厚度指数(MT%)、右心肥大指数(RVHI),采用Griess法检测血清NO水平,采用Western blotting法检测肺组织PI3K、AKT蛋白的相对表达量。结果PAH组MT%、RVHI及肺组织PI3K、AKT蛋白相对表达量均较正常组升高,但血清NO水平降低(P<0.05或<0.01);与PAH组相比,Cav1组、F92A-Cav1组中MT%、RVHI及肺组织PI3K、AKT蛋白相对表达量均降低,而血清NO水平增加,以F92A-Cav1组为著(P<0.05或<0.01)。结论 r BMSCs/F92A-Cav1可通过解除野生型Cav1对e NOS的抑制,促进NO释放,从而抑制PAH大鼠肺组织PI3K/AKT通路激活,进而发挥对PAH大鼠的治疗作用。

To investigate the therapeutic effect and mechanism of Caveolin-1 （Cav1） mutant to F92A-Cav1 modified rat bone marrow mesenchymal stem cells （rBMSCs/F92A-Cav1） on rats with pulmonary hypertension （PAH）.Methods PAH was induced by intraperitoneal injection of 1% monocrotaline （MCT, 60 mg/kg） in adult male Wistar rats.The PAH rats were randomly divided into four groups （10 rats/group） after 2 weeks of MCT injection： PAH group （only MCT）, Cav1 group （transduced with LV-Cav1 modified rBMSCs）, F92A-Cav1 group （transduced with LV-F92A-Cav1modified rBMSCs）, and the control group.Gene modified rBMSCs （1×106/ml） were transplanted into PAH rats in each group by tail vein injection.After 3 weeks of transplantation, the percentage of media wall thickness （MT%） and right ventricular hypertrophy index （RVHI） were evaluated by Image-Pro Plus 6 software, serum NO concentration was checked by Griess method, and the expression of phosphoinositide 3-kinase（PI3K） and protein kinase B （AKT） was detected by Western blotting.Results MT%, RVHI, PI3K and AKT was all increased but NO was decreased in the PAH group as compared with that of the control group（P〈0.05 or P〈0.01）.Compared with the PAH group, MT%, RVHI, PI3K and AKT was decreased but NO was increased in the Cav1 and F92A-Cav1 groups, especially in the F92A-Cav1 group （P〈0.01）.Conclusion F92A-Cav1 can abrogate the inhibitory effect of wild-type Cav1 on eNOS and promote the release of NO, then inhibit the activation of PI3K/Akt signaling pathway and thus exert its therapeutic effect on PAH rats.