人端粒酶反转录酶基因修饰人羊膜间充质干细胞移植治疗肺动脉高压

Transplantation of human telomerase reverse transcriptase-transfected human amniotic mesenchymal stem cells in the treatment of pulmonary hypertension

背景:随着干细胞移植治疗技术的发展及基因修饰技术的兴起为肺动脉高压的治疗提供了新的思路和方法。目的:观察人端粒酶反转录酶基因修饰人羊膜间充质干细胞移植对肺动脉高压大鼠的治疗效果。方法:66只成年雌性Wistar大鼠给予野百合碱腹腔注射(60 mg/kg)进行肺动脉高压模型复制,造模成功63只随机分成3组,其中肺动脉高压组于颈内静脉移植1 m L的L-DMEM培养基,人羊膜间充质干细胞组于颈内静脉移植1 m L人羊膜间充质干细胞悬液,人端粒酶反转录酶组于颈内静脉移植1 m L人端粒酶反转录酶转染人羊膜间充质干细胞悬液。移植后3周观察对比各组大鼠血流动力学、血浆内皮素1水平,以及右心室的肥大指数及心肌细胞凋亡情况。结果与结论:1治疗3周后各组间大鼠动脉血压差异无显著性意义(P>0.05);人端粒酶反转录酶组大鼠肺动脉收缩期压力和平均肺动脉压明显低于肺动脉高压组、人羊膜间充质干细胞组,差异有显著性意义(P<0.05)。2各组大鼠右心室的肥大指数差异无显著性意义(P>0.05)。3与肺动脉高压组及人羊膜间充质干细胞组相比,人端粒酶反转录酶组大鼠血浆内皮素1水平显著降低,差异有显著性意义(P<0.05)。4人端粒酶反转录酶组心肌细胞凋亡数较人羊膜间充质干细胞组和肺动脉高压组明显减少。5结果显示携带人端粒酶反转录酶基因的人羊膜间充质干细胞移植能够改善肺动脉高压大鼠的血流动力学异常状态,还可以保护机体血管内皮细胞,减少心肌细胞的凋亡。

BACKGROUND： The development of stem cell transplantation and genetic modification technology provides new ideas and methods for the treatment of pulmonary hypertension. OBJECTIVE： To investigate the therapeutic effects of transplantation of human telomerase reverse transcriptase（h TERT）-transfected human amniotic mesenchymal stem cells in pulmonary hypertension rats. METHODS： Human amniotic mesenchymal stem cells were cultured and purified in vitro, and then transfected with adenovirus-medicated h TERT. Sixty-six adult Wistar rats were enrolled to prepare pulmonary hypertension models through intraperitoneal injection of 60 mg/kg monocrotaline and then 63 model rats were randomly assigned into three groups： model group treated with transplantation of 1 mL of L-DMEM via the jugular vein, cell transplantation group treated with transplantation of 1 mL of untransfected human amniotic mesenchymal stem cell suspension, and transfection group treated with transplantation of 1 mL of human amniotic mesenchymal stem cell suspension transfected with h TERT. Hemodynamic changes, plasma endothelin-1 level, hypertrophy index of the right ventricle and myocardial cell apoptosis were compared among different groups at 3 weeks after transplantation. RESULTS AND CONCLUSION： After 3 weeks of treatment, there were no differences in the arterial blood pressure of the three groups（P〉0.05）; however, the systolic pressure of the pulmonary artery and mean pulmonary arterial pressure were significantly lower in the transfection group than the model group and cell transplantation group（P〈0.05）. Hypertrophy index of the right ventricle had no difference among the three groups（P〉0.05）. The level of plasma endothelin-1 was significantly lower in the transfection group than the model group and cell transplantation group（P〈0.05）. Apoptosis in myocardial cells was significantly reduced in the transfection group compared with the model group and cell transplantation group（P〈0.05）. Experimental findings suggest that the transplantation of h TERT-transfected human amniotic mesenchymal stem cells can improve the hemodynamic levels in pulmonary blood vessels of pulmonary hypertension rats to protect vascular endothelial cells and reduce myocardial cell apoptosis.