G-SCF联合HGF-MSCs治疗肺动脉高压大鼠的疗效研究

目的探讨重组粒细胞集落刺激因子(G-SCF)联合携带肝细胞生长因子(HGF)基因的骨髓间充质干细胞(MSCs)移植对野百合碱诱导的实验性肺动脉高压(PAH)大鼠的治疗效果。方法取3周龄雄性SD大鼠骨髓,经贴壁筛选法体外培养并纯化MSCs,利用腺病毒(Ad)介导的基因转染方法,将HGF基因导入MSCs。将50只SD大鼠随机分为5组(每组10只):PAH组,采用腹腔注射野百合碱(60mg/kg)复制PAH模型后,经颈内静脉移植1ml低糖伊戈尔培养基(L-DMEM);MSCs组,复制PAH模型后,移植5×106/L的空腺病毒载体转染的MSCs细胞悬液1ml;G-CSF组,经腹腔注射G-SCF[100μg/(kg.d)],连续5d;HGF组,移植5×106/L携带HGF基因的MSCs细胞悬液1ml;HGF+G-CSF组,移植5×106/L携带HGF基因的MSCs细胞悬液1ml并且经腹腔注射G-SCF[100μg/(kg.d),连续5d。21d后,观察各组血流动力学指标、右心室(RV)与左心室(LV)重量比值(RV/LV比值)、血管密度及血浆中内皮素-1(ET-1)的表达情况。结果 PAH大鼠治疗21d后,实验各组间全身动脉血压无明显差异;HGF+G-SCF组及HGF组中平均肺动脉压较其他各组显著下降(P<0.05);HGF组及HGF+G-SCF组RV/LV比值显著低于其他3组(P<0.05);HGF+G-SCF组血管密度高于其他各组,但差异无统计学意义(P>0.05);HGF组及HGF+G-SCF组血浆中ET-1的含量显著低于PAH组(P<0.05)。结论 G-SCF联合HGF-MSCs移植可有效减轻野百合碱诱导的大鼠PAH的进程,明显改善心功能及血流动力学指标,这些效应可能是通过增加肺血管数量及减少缩血管因子来实现的。

The effects of the rats with pulmonary arterial hypertension treated with HGF-MSCs and granulocyte colony-stimulating factor

Objective To study the effects of the combined therapy on monocrotaline-induced pulmonary artery hypertension in rat model.Methods MSCs obtained from bone marrow of three weeks Sprague-Dawley（SD） rats were cultured,purified and propagated in vitro.Then MSCs were transfected with Ad-HGF.The rats were randomly divided into 5 groups（n=10） ：PAH group：injection 1ml of L-DMEM into jugular veins after monocrotaline（MCT）,60 mg/kg,was injected by intraperitoneal injection to establish the models of PAH;MSCs group ：transplantation of 5 ×106 MSCs transfected with an empty adenovirus vector;HGF group：transplantation of 5×106MSCs transfected with Ad-HGF into jugular veins;G-CSF group：intraperitoneal injection G-CSF,100μg/kg daily for 5d;HGF＋G-CSF group：transplantation of 5×106 MSCs transfected with Ad-HGF into jugular veins and intraperitoneal injection G-CSF,100μg/kg daily for 5d.Three weeks later（21d）,the changes of pulmonary vasculature were detected.Results After the PAH rats were treated for 3 weeks the mPAP of HGF＋G-SCF and HGF group obviously descreased（P0.05）,but the BP wasn’t affected in the blood rheologic indexes.The weight ratio of RV/LV of HGF and HGF＋G-SCF group obviously declined（P0.05）.The number of capillary of HGF＋G-SCF group was the maximum,however the difference wasn’t significant.The ET-1 of HGF and HGF＋G-SCF group in plasma was evidently lower than PAH group（P0.05）.Conclusions The combined therapy of G-SCF and HGF-MSCs could effectively reduce the progression of PAH and obviously improve the heart function and the blood rheologic indexes by increas ing the number of capillary and decreasing the concentration of vasoconstrictor factor.