【摘要】 目的观察移植不同细胞数量的脂肪间充质干细胞(ADMSC)及移植后不同时间内ADMSC在野百合碱诱导的肺动脉高压(PAH)大鼠肺血管内的定植情况。方法实验分两部分:(1)不同时间细胞定植(时效):160只SD大鼠随机分为正常肺动脉压对照组1(Ctrl1)、肺动脉高压组1(PAH1)、正常肺动脉压移植组1(Ctrl1-ADMSC)和肺动脉高压移植组1(PAH1-ADMSC)。PAH1和PAH1-ADMSC腹腔注射野百合碱。Ctrl1-ADMSC和PAH1-ADMSC于野百合碱注射2周后移植1×10~6/mL ADMSC。各组分别于移植1、3、7、14和21d后测定肺动脉平均压(mPAP)。取肺组织切片并予滂胺天空蓝和二脒基苯基吲哚(DAPI)染色,观察ADMSC在肺血管的定植。(2)不同数量细胞移植与定植的关系(量效):80只SD大鼠随机分为正常肺动脉压移植组2(Ctrl2-A)、肺动脉高压移植组2(PAH2-A)。PAH2-A腹腔注射野百合碱。2周后各移植5个不同细胞数量(A0:0、A1:5×10~5、A2:1×10~6、A3:5×10~6、A4:1×10~7/mL)的ADMSC,21d后观察mPAP及ADMSC的定植。结果(1)PAH1-ADMSC中肺血管中定植的ADMSC呈时间依赖性减少[1~21d分别为(12.00±3.29)、(7.00±1.69)、(5.50±1.41)、(4.13±1.25)、(1.75±0.71)个]。与PAH1比较,PAH1-ADMSC的mPAP在移植7d后降低[(23.36±2.47)比(27.37±2.18)mm Hg,P<0.05]。(2)PAH2-A中肺血管定植的细胞数呈细胞数量依赖性增加[0、(3.13±1.25)、(7.00±2.00)、(13.25±1.91)、(22.00±8.05)个];对比移植细胞数为0的PAH大鼠,移植(1~5)×10~6时,PAH大鼠的mPAP显著降低[(28.6±3.0)、(26.0±1.7)比(39.8±1.8)mm Hg,P<0.05]。结论ADMSC可定植于肺血管内,并降低野百合碱诱导的PAH大鼠的mPAP;最佳移植数为(1~5)×10~6/mL。更多还原

【Abstract】 Objective To observe the colonization of adipose-derived mesenchymal stem cells(ADMSC)in the pulmonary vasculature of rats with monocrotaline(MCT)-induced pulmonary arterial hypertension(PAH)when using various numbers of transplanted cells,as well as at different times after the transplantation.Methods This study consisted of two parts:(1)160 Sprague-Dawley(SD)rats were randomly divided into the normal pulmonary arterial pressure control group(Ctrl1 group),pulmonary arterial hypertension group 1(PAH1 group),Ctrl1-ADMSC group and PAH1-ADMSC group.The rats in the PAH1 and PAH1-ADMSC groups were administered MCT by intraperitoneal injection.1×10~6/mL of ADMSC were transplanted to the rats in the Ctrl1-ADMSC and PAH1-ADMSC groups after 2 weeks of the injection of MCT.Mean pulmonary arterial pressure(mPAP)was measured in each group after 1,3,7,14,and 21 days of the transplantation,respectively.The lung tissue slices were stained with pontamine sky blue and diamidino-phenylindole(DAPI),and then the colonization of ADMSC in the pulmonary vessels was observed.(2)80SD rats were randomly divided into the Ctrl2-A group(normal pulmonary arterial pressure-transplantation group 2)and PAH2-A group(PAH-transplantation group 2).The rats in the PAH2-A group were administered MCT by intraperitoneal injection.Five different numbers of ADMSC(A0:0,A1:5×10~5,A2:1×10~6,A3:5×10~6,A4:1×10~7/mL)were transplanted to the two groups after 2 weeks.The mPAP was measured and the colonization of ADMSC was observed after 21 days of the transplantation.Results The colonization numbers of ADMSC in the pulmonary vessels of the PAH1-ADMSC group were time-dependently reduced(12.00±3.29,7.00±1.69,5.50±1.41,4.13±1.25,1.75±0.71after 1,3,7,14,and 21days of the transplantation,respectively).Compared with the PAH1group,the mPAP in the PAH1-ADMSC group was decreased after 7 days of the transplantation[(23.36±2.47)vs(27.37±2.18)mm Hg,P<0.05].Compared with the Ctrl2-A group,the colonization numbers of ADMSC in the pulmonary vessels of the PAH2-A group were increased with an increase in the number of transplanted cells(0,3.13±1.25,7.00±2.00,13.25±1.91 and 22.00±8.05,respectively).Compared with the PAH group with no cell transplantation,the mPAP in the PAH groups transplanted(1-5)×10~6/mL of ADMSC was significantly decreased[(28.6±3.0),(26.0±1.7)vs(39.8±1.8)mm Hg,respectively,all P<0.05].Conclusion The transplanted ADMSC could colonize in the pulmonary vasculature and could reduce mPAP in the rats with MCT-induced PAH,and the optimum concentration of ADMSC was(1-5)×10~6/mL.更多还原