华通胶间充质干细胞移植改善大鼠肺动脉高压的疗效观察

Transplantation ofWharton’s jelly derived-mesenchymal stem cells Attenuates Pulmonary Arterial Hypertension in Rats

目的研究华通胶间充质干细胞(wharton’s jelly derived-mesenchymal stem cells,WJ-MSCs)移植对野百合碱(monocrotaline,MCT)诱导的大鼠肺动脉高压(pulmonary arterial hypertension,PAH)模型的治疗效果。方法在无血清培养条件下体外培养WJ-MSCs并进行细胞表型鉴定。30只雄性SD大鼠随机分为对照组、MCT组和WJMSC移植组。通过腹腔注射野百合碱制备大鼠PAH模型,2周后WJMSC移植组大鼠从尾静脉注射WJ-MSCs细胞悬液,其余组注射等量生理盐水。3周后对比三组大鼠生存状态、右心室收缩压(right ventricular systolic pressure,RVSP)、右心肥大指数(right ventricle hypertrophy index,RVHI),肺组织病理观察肺小动脉形态,分析比较中膜厚度占血管外径百分比、管腔狭窄率。结果无血清培养体系获得的WJ-MSCs表达CD44、CD73、CD90、CD105,而不表达CD45、CD34。与对照组及WJMSC移植组相比,MCT组大鼠病死率高,体重增长缓慢;RVSP、RVHI、中膜厚度占血管外径百分比、管腔狭窄率明显升高(P<0.01);与MCT组相比,WJMSC移植组大鼠RVSP(P<0.01)、RVHI(P<0.05)、中膜厚度占血管外径百分比(P<0.01)、管腔狭窄率(P<0.01)显著改善。结论无血清体系培养的WJ-MSCs符合间充质干细胞的表型特点。WJ-MSCs移植可显著降低PAH大鼠肺动脉压力,改善右心肥厚,缓解肺血管重构。

Objective To investigate the therapy effect of Wharton’s jelly derived-mesenchymal stem cells(WJMSCs)in pulmonary arterial hypertensive rats induced by monocrotaline.Methods WJ-MSCs were cultured in serumfree culture medium in vitro and the cell phenotypes were identified.Thirty adult male Sprague-Dawley rats were divided into control group,MCT group and WJMSC group randomly.Sprague-Dawley rats were intraperitoneally injected with monocrotaline to induce pulmonary arterial hypertension(PAH).After two weeks,rats in the WJMSC group were injected with WJMSCs via tail vein and rats in the other two groups were injected with saline.Three weeks after transplantation,right ventricular systolic pressure(RVSP),right ventricle hypertrophy index(RVHI),pathological morphology of pulmonary arterioles,percentage of medial thickness,and percentage of membrane area of pulmonary arterioles were evaluated.Results WJ-MSCs cultured in serum-free medium were positive for CD44,CD73,CD90,CD105 and negative for CD45,CD34.Compared with the control group and WJMSC group,the mortality of rats in MCT group was higher and the body weight increased slowly.Meanwhile,RVSP,RVHI,percentage of medial thickness,and percentage of membrane area of pulmonary arterioles were significantly increased(P<0.01).Compared with the MCT group,RVSP(P<0.01),RVHI(P<0.05),percentage of medial thickness(P<0.01),and percentage of membrane area of pulmonary arterioles(P<0.01)in the rats of WJMSC group were notably ameliorated.Conclusions The phenotypes of WJ-MSCs cultured in serum-free medium were consistent with characteristics of mesenchymal stem cells.Transplantation of WJMSCs could significantly decrease pulmonary artery pressure,ameliorate right heart hypertrophy and pulmonary vascular remodeling in PAH rats.