骨髓间充质干细胞移植治疗实验性肺动脉高压大鼠血流动力学指标的变化

Hemodynamic Parameter Changes in Pulmonary Arterial Hypertension Treated by Transplantation of Bone Marrow Mesenchymal Stem Cells in Experimental Rats

目的探讨骨髓间充质干细胞(MMSCs)移植治疗大鼠实验性肺动脉高压(PAH)后血流动力学指标的变化。方法取SD大鼠骨髓,经贴壁筛选法体外培养并纯化MMSCs,移植前采用Hoechst33342荧光染料标记。SD大鼠90只随机分为3组(n=30):MMSCs移植组(M组),单纯PAH组(H组)和正常对照组(N组)。M组采用野百合碱(50mg/kg)注射复制PAH模型成功后21d经舌下静脉移植5×109L-1的MMSCs细胞悬液1mL;H组复制PAH模型后21d移植1mL低糖伊格尔培养基(L-DMEM)细胞培养液;N组注射9g/L盐水(6mL/kg)。饲养28d,观察各组动物动脉血气指标、右室收缩压(RVSP)、右心指数及肺小动脉血管的变化。结果H组大鼠注射野百合碱21d后RVSP、右心指数、管壁中膜厚度(MT)显著增高,但管腔面积(VA)与血管总面积(TAA)比值(VA%)及动脉血pH值、pa(O2)、HCO3-均显著下降;M组移植MMSCs28d后与H组比较,RVSP显著降低[(32.20±2.32)mmHgvs(48.30±1.56)mmHgP<0.05],右心指数下降[(38.80±3.24)%vs(45.10±3.43)%P<0.05],M组血气分析结果显示pH值、pa(O2)、HCO3-值接近N组。荧光显微镜观察到M组荧光标记的MMSCs在肺内定植,且分化成大量的侧支循环,肺动脉重构得到有效逆转。结论MMSCs移植可有效减轻甚至逆转野百合碱诱导的大鼠PAH进程,明显改善其心功能及血流动力学指标。

Objective To explore hemodynamic parameter changes in pulmonary arterial hypertension（PAH） treated by transplantation of bone marrow mesenchymal stem cells（MMSCs） in the experimental rats. Methods MMSCs cells were collected from bone marrow of Spra- gue -Dawleye（SD） rat＇s femoral and tibial bones,cultured and passaged in vitro,then stained by Hoechst 33342 fluorescence dye stuff. Nine- ty healthy male SD rats were randomly divided into 3 groups （ n = 30 ） ： normal control group （ group N ）, MMSCs transplanted group （ group M） ,PAH model group （group H）. The rats in the two latter groups were given a single subcutaneous erotaline（50 mg/kg） to establish the model of PAH. The rats of group N were injected respectively a single subcutaneous 9 g/L saline water （6 mL/kg）. After 21 days,5 ×10^9 L-1 MMSCs cultured in 1 mL phosphate - buffered saline were infused into the rats respectively in group M by sublingual vein and 1 mL L - DMEM was given in group H; The indexalso of right ventricle systolic pressure（RVSP） ,right ventricle hypertrophy index ,arterial blood gas a- nalysis and the changes of small pulmonary blood vessel were observed after 28 days. Results The administration of MMSCs 28 days after PAH nearly completely prevented the increase in RVSP with PAH alone [ （32.20 ± 2.32） mmHg vs （48.30 ± 1.56） mmHg P 〈 0.05 ], right ventricle hypertrophy indexalso degraded[ （38.80 ±3.24）% vs （45.10±3.43） % P〈0.05] ,and relative media thickness raised obviously but the percentage of vessel area（ VA% ） decreased, results of blood gas analysis such as pH ,pa（O2 ）, HCO3 -degraded significant- ly. MMSCs labeled Hoechst 33342 were observed and were permanently planted and differentiated into plentiful collateral circulations in lung by fluorescence microscope. The pulmonary arteries remodeling was attenuated effectively in MMSCs group M. Conclusions These findings indicate that MMSCs might reduce and even reversion the progression of PAH induced by crotaline. And the heart function and hemodynamic parameter can be improved obviously.