芪丹通脉片对移植后骨髓间充质干细胞分化为内皮细胞的协同作用

Synergistic effect of Qidantongmai Tablet on differentiation of transplanted MSCs into endothelial cells

目的探讨芪丹通脉片对大鼠骨髓间充质干细胞(MSCs)移植后在缺血心肌组织中分化为内皮细胞的协同作用。方法采用密度梯度离心法分离和纯化大鼠MSCs,培养扩增至第3代后,应用5-溴脱氧尿嘧啶(BrdU)标记细胞。将16只SD大鼠随机分为两组:芪丹通脉片组(8只)和生理盐水对照组(8只),分别灌服芪丹通脉片浸膏干粉溶液2g生药/kg及等体积的生理盐水,每天1次,连续14天。结扎大鼠冠状动脉左前降支,制备急性心肌梗死模型,将BrdU标记的异体MSCs以微量注射的方法移植入缺血心肌内。4周后,用免疫荧光法鉴定移植后细胞表面血管生成素受体2(Tie2)的表达,并以CD31阳性细胞计算和比较两组的微血管密度(MVD)。结果芪丹通脉片组和生理盐水对照组大鼠左心室均发现BrdU阳性细胞(即植入细胞),而芪丹通脉片组BrdU阳性细胞中Tie2阳性信号表达程度均多于生理盐水对照组,芪丹通脉片组MVD大于生理盐水对照组。结论芪丹通脉片具有促进MSCs在缺血心肌中转化为内皮细胞,增加毛细血管密度的作用。

Objective To investigate the synergistic effect of Qidantongmai Tablet on differentiation of transplanted marrow mesenchymal stem cells（MSCs） into endothelial cells in the ischemic myocardium of rat. Methods Rat MSCs were isolated by density gradient method, cultured and amplified through 3 passages, then the cells were labeled with BrdU. Sixteen SD rats were divided into two groups randomly, the Qidantongmai Tablet group and NS control group. The suspension of Qidantongmai Tablet （2 g/kg） and NS were administered by gastrogavage for 14 days. The left anterior descending coronary artery of rat was ligated to establish animal model of acute myocardial infarction. The MSCs labeled with BrdU were implanted intramyocardially into the infarct area. After 4 weeks, the expression of Tie2 on transplanted MSCs was assessed. Microvessel density（MVD） was calculated and compared by CD31 positive cells. Results The BrdU positive cells were found in both groups. But the proportion of Tie2 positive cells was higher in Qidantongmai Tablet group than in NS control group, so as the MVD. Conclusion Qidantongmai Tablet has the synergistic effect on differentiation of transplanted MSCs into endothelial cells in the ischemic myocardium and enhances the MVD.