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目的探讨骨髓间充质干细胞(MSCs)移植对野百合碱诱导的肺动脉高压(PAH)大鼠肺血管重构的影响。方法体外分离、培养并纯化SD大鼠骨髓MSCs。健康雄性SD大鼠随机分为3组:正常对照组(n=15)、PAH组(n=20)和MSCs移植组(n=20)。PAH组和MSCs移植组大鼠一次性项背部皮下注射野百合碱(50 mg.kg-1),复制PAH肺血管重建动物模型。3组大鼠在同等条件下饲养。3周后,MSCs移植组大鼠经舌下静脉注射5×109 L-1的经Hoechst 33342标记的MSCs细胞悬液1 mL,PAH组大鼠予等量低糖-Dulbecco改良Eagle’s培养液(L-DMEM)注射,正常对照组不做任何处理。移植28 d,观察3组大鼠肺小动脉的显微结构及超微结构的改变。结果 PAH大鼠用MSCs移植28 d后,肺小动脉管壁厚度指标(WD/TD%、VA%)、放射性肺泡计数、肺非肌性小动脉肌化程度均较PAH组显著改善(P<0.01);其肺小动脉的重构及超微结构的血气屏障、线粒体、板层小体等改变均有明显改善。荧光显微镜观察到Hoechst 33342标记的MSCs在肺内定植,且分化成大量新生血管并形成侧支循环,肺动脉重构得到有效逆转。结论 MSCs移植可有效减轻并逆转肺动脉重构的进程,其作用机制是MSCs分化形成新生血管,建立了侧支循环,从而修复野百合碱诱导的肺损伤。
Objective To investigate the effects of bone marrow mesenchymal stem cells (MSCs) transplantation on pulmonary vascular remodeling induced by monocrotaline-induced pulmonary hypertension (PAH) in rats. Methods The bone marrow MSCs of SD rats were isolated, cultured and purified. Healthy male Sprague-Dawley rats were randomly divided into 3 groups: normal control group (n = 15), PAH group (n = 20) and MSCs transplantation group (n = 20). Rats in PAH group and MSCs transplantation group were injected subcutaneously with monocrotaline (50 mg · kg-1) on the back of the rats to replicate the pulmonary vascular reconstruction model of PAH. Three groups of rats were housed under the same conditions. Three weeks later, 1 mL of 5 × 109 L-1 Hoechst 33342-labeled MSCs was injected into the subconjunctival vein of MSCs transplanted group. The rats in PAH group were treated with the same amount of low-glucose-Dulbecco’s modified Eagle’s medium (L- DMEM) injection, the normal control group without any treatment. Transplanted for 28 days, the microstructure and ultrastructure of the pulmonary arterioles in the three groups were observed. Results After the MSCs were transplanted for 28 days in PAH rats, the thickness of pulmonary arterioles (WD / TD%, VA%), radioactive alveolar count and muscular pulmonary arterial noninvasive pulmonary artery were significantly improved (P < 0.01). The remodeling of pulmonary arterioles and ultrastructure of blood gas barrier, mitochondria, lamellar bodies and other changes were significantly improved. Fluorescence microscopy showed that Hoechst 33342-labeled MSCs colonized the lungs and differentiated into a large number of neovascular vessels and formed collateral circulation. Pulmonary artery remodeling was effectively reversed. Conclusion MSCs transplantation can effectively reduce and reverse the process of pulmonary artery remodeling. The mechanism is that MSCs differentiate to form new blood vessels and establish collateral circulation to repair monocrotaline-induced lung injury.