摘要
研究背景：干细胞移植治疗心肌梗死是近年来研究的热点,前期研究发现移植的干细胞能在受损心肌组织内存活、分化并改善心功能,可能的机制为移植细胞通过自分泌或旁分泌作用释放细胞因子,促进心肌细胞关键酶或动员内源性干细胞的归巢和分化,干细胞移植改善心肌细胞Na+-K+-ATP酶和Ca2+-ATP酶的活性已有研究,但能否促进外周血干细胞的归巢尚未见报道。目的：通过对心肌梗死大鼠模型局部注射移植同种异体骨髓间充质干细胞,观察能否改善心功能；在此基础上静脉注射同种异体外周血干细胞,观察能否进一步改善心功能,同时了解局部注射移植的骨髓间充质干细胞能否募集外周血干细胞。方法：32只体重为(180g-200g)的SD大鼠,雌雄不限,每组8只,随机分为四组：正常对照组(normal group)、假移植组(fake transplanted group)、骨髓间充质干细胞移植组(MSCs group)、骨髓间充质干细胞移植+内皮祖细胞移植组(MSCs+EPCs group)；分别于移植前、移植后5周测定大鼠心功能；免疫组化法检测梗死区标记的干细胞及骨髓间充质干细胞对外周血内皮祖细胞的募集情况。采用SPSS13.0软件进行统计学分析。结果：(1)移植前、移植后5周比较,正常组无明显差别(P>0.01),假移植组最大左室收缩末压(LVSP)、左室内压最大(最小)变化速率(LV士dp／dt max)于移植后5周均减小(P<0.01),而左室舒张末压(LVDP)上升(P<0.01),说明假移植组5周后心功能有恶化；MSCs组和MSCs+EPCs组在移植后5周最大左室收缩末压(LVSP)、左室内压最大(最小)变化速率(LV士dp／dt max)升高,有显著意义(P<0.01),而左室舒张末压(LVDP)降低(P<0.01),说明干细胞移植后心功能有一定程度的恢复；(2)干细胞移植后5周,与正常组相比,假移植组最大左室收缩末压(LVSP)、左室内压最大(最小)变化速率(LV士dp／dt max)均明显下降(P<0.01),而左室舒张末压(LVDP)明显升高(P<0.01)表明心肌梗死后心功能的恶化；MSCs组和MSCs+EPCs组大鼠的LVSP、LV士dp／dt max下降(P<0.01),而LVDP明显升高(P<0.01),表明干细胞移植后心功能有一定程度的恢复,但尚未达到正常；与假移植组相比,MSCs组和MSCs+EPCs组大鼠的LVSP.LV士dp／dt max均有明显升高(P<0.01),而LVDP明显降低(P<0.01)表明细胞移植后心功能优于假移植组；而MSCs组与MSCs+ECPs组相比,其LVSP、LV士dp／dt max、LVDP改变均不明显(P>0.01),两组比较无统计学意义,说明其改善心功能效果和骨髓间充质干细胞移植组相当。(3)免疫荧光染色结果显示移植后5周明显可见梗死区组织BrdU标记的干细胞,未见DAPI标记的干细胞,表明骨髓间充质干细胞对外周血干细胞无募集作用。结论：①大鼠心肌梗死模型的成功建立。②干细胞移植后较移植前心功能有改善。③MSCs+EPCs组与MSCs组相比,进一步改善心功能不明显。④骨髓间充质干细胞对内皮祖细胞没有募集作用。
Abstract:Background:Recent years, stem cells transplantation was one of the focus in cardiology. The study found that transplanted stem cells can survive、differentiation and improve heart function in damaged tissue, Possible mechanism was transplanted cells promote key enzymes or the nest and endogenous stem cell differentiation through the transplanted cells secrete or paracrine effect. Stem cells improved Na+-K+-ATP enzymes and Ca2+-ATP enzymes which we had researched, but it did not know if they can promote the nest. Objective:To observe whether they could improve heart function and recruit peripheral blood stem cells after local injected homogeneous bone marrow mesenchymal stem cells to myocardial infarction rats model and then intravenous peripheral blood stem cell. Methods:Thirty-two SD rats (180-200g) were randomly divided into 4 groups (8 rats each group), ie. normal group:did not made model; fake transplanted group:made myocardial infarction operation but injected DMEM; MSCs transplanted group: partly injected MSCs on myocardial infarction rats model; MSCs and EPCs transplanted group:partly injected MSCs and peripheral blood stem cell on myocardial infarction rats model. Respectively detected heart function at pre-and post-transplanted 5 weeks; Immunohistochemistry method to detect marked stem cells and whether bone marrow mesenchymal cells could raise peripheral endothelial progenitor cells. Analyzed these index through SPSS 13.0 statistical software. Result:(1) Comparison between pre-and post-transplantation of stem cell, normal group did not changed;fake transplanted groups all decreased in LVSP、LV±dp/dt max and increased in LVDP after 5 weeks (p<0.01),this indicated the worsen heart function; MSCs transplanted group and MSCs +EPCs transplanted group had improved in LVSP、LV±dp/dt max (p<0.01),but did not obviously in LVDP, this indicated the recovery heart function after 5weeks;(2) After 5weeks of stem cells transplanted, compared to normal group, a significant decreased in LVSP、LV±dp/dt max (p<0.01) and increased in LVDP (p<0.01) in fake transplanted group, this indicated myocardial got worse; MSCs transplanted group and MSCs+EPCs transplanted group also had decreased in LVSP、LV±dp/dtmax and increased in LVDP(p<0.01),this indicated the recovery of myocardial,but did not reach normal; compared to fake transplanted group, MSCs group and MSCs+EPCs transplanted group had significant increased in LVSP、LV±dp/dtmax (p<0.01) and decreased in LVDP (p<0.01), this indicated stem cells transplanted group better than fake group; compared between stem cells transplanted group, there did not obviously change in LVSP、LV±dp/dt max and LVDP (p>0.01),this indicated the small difference of them. (3)5 weeks later we detected BrdU marker at infarction areas, but did not detect DAPI marker,this indicated they do not gather. Conclusion:①Successfully established the model of myocardial infarction②There had obviously improved at 5weeks later③Did not obviously improved in MSCs+ EPCs transplanted group④Bone marrow mesenchymal cells did not raise endothelial progenitor cells.
引文
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