何志旭,汪浩文,尚峰,严虎,杨燕,崔冬冰
HE Zhi-xu,WANG Hao-wen,SHANG Feng,YAN Hu,YANG Yan,CUI Dong-bing(Research Center of Stem Cell

• 1:贵阳医学院组织工程及干细胞研究中心
• 2:贵阳医学院附院儿科

摘要(Abstract)：

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)的疗效及血流动力学指标的变化。方法:取SD大鼠的骨髓,经贴壁筛选法体外培养并纯化MMSCs,移植前采用Hoechst 33342荧光染料标记;实验动物随机分为3组:MMSCs移植组(M组),单纯肺动脉高压组(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3周,经舌下静脉移植5×106/ml的MMSCs细胞悬液1ml;H组复制PAH模型后3周,移植1mlL-DMEM细胞培养液,C组不作任何处理;饲养4周后,观察各组动物动脉血气指标、右心室收缩压、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,右心室收缩压、右心指数、管壁中膜厚度明显增高,但管腔面积与血管总面积比值及动脉血pH值、PaO2、HCO3-明显下降;M组移植MMSC28d后,较H组右心室收缩压明显降低[移植组为(32.0±2.32)mmHg,模型组为(48.30±1.56)mmHg],右心指数下降[移植组为(38.80±3.24)%,模型组为(45.10±3.43)%,两组相比P<0.05],血气分析结果显示M组pH值、PaO2、HCO3-值接近正常对照组;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枝循环,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肺动脉高压的进程,明显改善心功能及血流动力学指标。
Objective:To explore the change of hemodynamic parameter in bone marrow mesenchymal stem cells(MMSCs) transplantation to treat the experimental pulmonary arterial hypertension (PAH) in rats. Method:MMSCs cells were collected from bone marrow of SD rat's femoral and tibial bones,cultured and passaged in vitro,then purified by density gradient centrifugation and adhesive-screening method.60 healthy male Sprague-Dawleye rats(mean weighing 180±5 g) were randomly divided into 3 groups:normal control group (group C),MMSCs transplanted group(group M),PAH model group (group H ).The rats of the two latter group were given a single subcutaneous crotaline(50 mg/kg) to make the model of PAH. The rats of normal control group were injected respectively a single subcutaneous 0.9%NaCL(6 ml/kg).The three groups were fed in the same condition.After 3 weeks,5×106 MMSCs cultured in l ml phosphate-buffered saline were infused into the rats respectively in group M by sublingual vein and 1 ml L-DMEM was did in group H.The index of right ventricle systolic pressure(RVSP),right ventricle hypertrophy index,arterial blood gas analysis and the changes of small pulmonary blood vessel were observed after 28 day.Results:The administration of MMSCs 28 days after PAH the percent of viability is 100 in transplanted group,and it nearly completely prevented the increase in right ventricular systolic pressure(RVSP)with PAH alone (32.20±2.32 versus 48.30±1.56 mm Hg,respectively P<0.05),right ventricle hypertrophy index degraded too(38.80±3.24% versus 45.10±3.43 %,respectively P<0.05 ),and relative media thickness(MT) decreased obviously but the VA% raised,results of Blood gas analysis such as pH、PaO2、HCO3-degraded significantly. MMSCs labeled Hoechst 33342 were observed that they can permanent planted and differentiated into plentiful collateral circulations in lung by fluorescence microscope. The pulmonary arteries remodeling was attenuated effecfively in MSCs transplanted group. Conclusions:These results indicated that bone marrow–derived Mesenchymal stem cells (MMSCs) could reduce and even reversion the progression of Pulmonary arterial hypertension (PAH) induced by crotaline.The heart function and hemodynamic parameter had obvious improved.

关键词(KeyWords)： 高血压,肺性;骨髓间充质干细胞;血气分析;血流动力学
hypertension,pulmonary; mesenchymal stem cells; blood gas analysis; hemodynamics

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30560159);; 教育部新世纪优秀人才支持计划项目(NCET-05-0820);; 贵州省高校自然科学基金资助项目(黔科教2005-7号)

作者(Author): 何志旭,汪浩文,尚峰,严虎,杨燕,崔冬冰
HE Zhi-xu,WANG Hao-wen,SHANG Feng,YAN Hu,YANG Yan,CUI Dong-bing(Research Center of Stem Cell

DOI: 10.19367/j.cnki.1000-2707.2008.06.003

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