肌肉注射异种脐带间充质干细胞大鼠心肌肌钙蛋白I及血清相关因子的表达

Effects of intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells on myocardial cardiac troponin and serum-related factors in rats

背景:目前关于肌肉注射异种脐带间充质干细胞是否安全缺乏理论依据。目的:观察异种脐带间充质干细胞肌肉注射对大鼠心肌血管内皮生长因子、肌钙蛋白I及血清血管内皮生长因子、肝细胞生长因子、胰岛素样生长因子1和粒-巨噬细胞集落刺激因子表达的影响。方法:正常Wistar大鼠分为6组,分别于大鼠四肢肌肉肌注磷酸盐缓冲液、DMEM液、培养人脐带间充质干细胞上清液、0.25×105,1.0×105和4.0×105人脐带间充质干细胞。4周后重复注射,8周后取血和心肌组织,ELISA法检测血清血管内皮生长因子、肝细胞生长因子、胰岛素样生长因子1和粒-巨噬细胞集落刺激因子水平,免疫组化法检测大鼠心肌肌钙蛋白I及血管内皮生长因子表达情况。结果与结论:肌注前后各组间大鼠血清血管内皮生长因子、肝细胞生长因子、胰岛素样生长因子1和粒-巨噬细胞集落刺激因子水平差异无显著性意义(P>0.05);同组大鼠肌注前后血清血管内皮生长因子、肝细胞生长因子、胰岛素样生长因子1和粒-巨噬细胞集落刺激因子水平差异亦无显著性意义(P>0.05)。肌注后8周,各组大鼠肌钙蛋白I在心肌细胞胞浆中均匀分布且呈强阳性表达;血管内皮生长因子亦在心肌细胞胞浆中分布,但呈弱阳性表达;各组间大鼠心肌组织肌钙蛋白I及血管内皮生长因子水平比较差异无显著性意义(P>0.05)。提示肌注人脐带间充质干细胞或其培养上清液对正常大鼠血清血管内皮生长因子、肝细胞生长因子、胰岛素样生长因子1、粒-巨噬细胞集落刺激因子水平及心肌组织肌钙蛋白I及血管内皮生长因子的表达无影响。

BACKGROUND：Current researches on whether the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells （UC-MSCs） is safe or not lack theoretical basis. OBJECTIVE：To explore the effects of intramuscular injection of heterogeneous UC-MSCs on expression of myocardial vascular endothelial growth factor （VEGF）, cardiac troponin I （cTnI） and serum VEGF, hepatocyte growth factor （HGF）, insulin-like growth factor 1 （IGF-1） and granulocyte macrophage colony stimulating factor （GM-CSF） in normal Wistar rats. METHODS：A total of 60 Wistar rats were divided into six groups randomly. Rats in the six groups were respectively administrated with intramuscular injection of PBS, Dulbecco’s modified Eagle’s medium, hUC-MSC supernatant, 0.25×10^5;105, 1.0×10^5;105, 4.0×10^5;105 hUC-MSCs. Rats received a second intramuscular injection 4 weeks after first injection. Eight weeks later, the blood sample and myocardial tissue were taken. The serum concentration of VEGF, HGF, IGF-1 and GM-CSF were measured with enzyme-linked immunosorbent assay kit. The myocardial VEGF and cTnI expression were detected by immunohistochemical technique. RESULTS AND CONCLUSION：There was no significant difference in the serum concentration of the VEGF, HGF, IGF-1 and GM-CSF among the groups before and after injection （P〉0.05）. In the same group, no statistical significant＆amp;nbsp;changes in the serum concentration of the VEGF, HGF, IGF-1 and GM-CSF were found among the rats before and after injection （P〉0.05）. Eight weeks after injection, weak positive expression of the VEGF in the cytoplasm of cardiocytes in the six groups was observed, and strong positive expression of the cTnI in the cytoplasm of cardiocytes in the six groups was observed. There was no significant difference in the VEGF and cTnI content in the myocardium among the groups （P〉0.05）. Intramuscular injection of hUC-MSCs or the supernatant of hUC-MSCs had no effects on the serum concentration of the VEGF, HGF, IGF-1, GM-CSF and the myocardial VEGF and cTnI expression in normal Wistar rats.