摘要目的 观察骨髓间充质干细胞(MSCs)静脉移植对野百合碱(MCT)诱导肺动脉高压(PAH)大鼠的内皮素-1(ET-1)表达的影响.方法 培养大鼠幼鼠骨髓MSCs,传代纯化,取第3～5代细胞进行移植.Wistar大鼠30只,成组设计(n=10)为3组:实验组、MCT组和对照组.前两组腹腔注射60 mg/kg MCT(对照组注射等量生理盐水)诱导后,实验组颈外静脉注射含1×106 MSCs的磷酸盐缓冲液(PBS),后两组注入等量PBS.4周后检测大鼠右心室收缩压(RVSP)；肺组织苏木素-伊红(HE)染色；肺组织分离mRNA,逆转录酶聚合酶链反应(RT-PCR)法检测组织表达前体原ET-1 mRNA的水平；酶联免疫吸附试验(ELISA)法测定血清ET-1的表达.统计数据进行成组t检验.结果 MSCs颈外静脉移植4周后,实验组与MCT组比较RVSP明显下降[(35.6±8.4) mmHg(1 mm Hg =0.133 kPa)比(47.2±10.5) mmHg,P＜0.05]；病理染色可见实验组与对照组比较肺小动脉壁明显变薄；肺组织前体原ET-1 mRNA半定量显示实验组较MCT组表达减少(0.251 ±0.048比0.391 ±0.035,P＜0.05),血浆ET-1结果均显示对照组ET-1表达很少(2.23 ±0.43) ng/L,与MCT组比较,MSCs移植降低了ET-1的表达[(4.05±0.82) ng/L比(5.75 ±0.75) ng/L,P＜0.05].结论 MSCs颈外静脉移植可以抑制MCT对大鼠肺损伤作用,减少ET-1的mRNA及血清ET-1的表达.

abstractsObjective To observe changes of endothelin-1 ( ET-1 ) in rats with pulmonary arterial hypertension (PAH) induced by monocrotaline (MCT) after mesenchymal stem cells (MSCs) transplantation.Methods MSCs of young Wistar rats were cultured,purified,and those at the third to fifth generation were prepared for transplantation.Thirty male Wistar rats were divided into three group ( n =10 each) randomly:experimental group,MCT group and control group.The rats were injected intraperitoneally with 60 mg/kg MCT in the former two groups,but with normal saline in control group,respectively.Meanwhile,1 × 106 MSCs in 1 ml phosphate belanced solution (PBS) were transplanted into rats through external jugular vein in experimental group,but only PBS injected in the latter two groups respectively.At the 4th week after transplantation,the right ventricular systolic pressure (RVSP) was measured and the lung tissues were stained by hematoxylin-eosin staining.The expression of prepro-ET-1 mRNA was observed by reverse transcription polymerase chain reaction (RT-PCR) in the lungs and the serum endothelin-1 levels were detcrminated by enzyme linked immunosorbent assay (ELISA).All data were analyzed statistically by independent t test.Results At 4th week after MSCs transplantation,RVSP was significantly decreased in experimental group as compared with MCT group [(35.6 ± 8.4) mm Hg (1 mm Hg=0.133 kPa) vs (47.2 ± 10.5) mm Hg,P ＜0.05].Pathological staining revealed that the media thickness of the pulmonary arterioles was thinner in experimental group than in MCT group.The prepro-ET-1 mRNA expression in the lung tissues was decreased significantly in experimental group as compared with MCT group (0.251 ±0.048 vs 0.391 ± 0.035,P ＜ 0.05).The plasma ET-1 levels were low (2.23 ± 0.43 ) ng/L in control group.MSCs transplantation could significantly decrease the expression of ET-1 as compared with MCT group [(4.05 ±0.82) ng/L vs (5.75 ±0.75) ng/L,P＜0.05].Conclusion Intravenous MSCs transplantation can prevent lungs injury induced by MCT and reduce both prepro ET-1 mRNA expression in lungs and ET-1 peptide levels in plasma.