【摘要】 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。更多还原

【Abstract】 Objective To investigate the therapeutic effect and mechanism of Caveolin-1( Cav1) mutant to F92ACav1 modified rat bone marrow mesenchymal stem cells( r BMSCs/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 r BMSCs),F92A-Cav1 group( transduced with LV-F92A-Cav1 modified r BMSCs),and the control group. Gene modified r BMSCs( 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,PI3 K 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,PI3 K 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 e NOS and promote the release of NO,then inhibit the activation of PI3K/Akt signaling pathway and thus exert its therapeutic effect on PAH rats.更多还原