肌肉注射脐带间充质干细胞对扩张型心肌病大鼠相关细胞因子的影响

Human umbilical cord mesenchymal stem cells via intramuscular injection influence the expression of cytokines related to dilated cardiomyopathy in rats

背景:已有研究表明,在一定的剂量范围内,健康大鼠肌肉注射异种脐带间充质干细胞安全可靠,也证实了这一途径对扩张型心肌病大鼠心力衰竭同样安全有效。目的:探讨肌肉注射脐带间充质干细胞对扩张型心肌病大鼠相关细胞因子表达的影响。方法:雄性SD大鼠160只,随机抽取20只作为正常组,余大鼠通过阿霉素腹腔注射方式建立扩张型心肌病模型。造模成功大鼠随机分为模型组、上清液祖、低剂量组、中剂量组、高剂量组,分别经四肢肌肉注射人脐带间充质干细胞培养液、上清液及不同剂量的人脐带间充质干细胞,注射后观察大鼠一般情况,并于首次注射后4周再次给予相同剂量的培养液、上清液或脐带间充质干细胞。结果与结论:ELISA检测结果显示,肌注前后模型组大鼠血清肝细胞生长因子、白血病抑制因子、粒细胞集落刺激生物因子及血管内皮生长因子水平均较正常组增高(P<0.05),肌注后低剂量组肝细胞生长因子、白血病抑制因子、血管内皮生长因子和粒细胞集落刺激生物因子较肌注前显著升高(P<0.05),且较模型组明显升高(P<0.05),中剂量组白血病抑制因子较肌注前明显升高(P<0.05),高剂量组肝细胞生长因子、血管内皮生长因子和粒细胞集落刺激生物因子较肌注前无明显差异(P>0.05)。免疫组化及RT-PCR结果均显示,各肌肉注射组大鼠心肌细胞胰岛素样生长因子1、血管内皮生长因子、肝细胞生长因子均较正常组表达增强,中剂量组呈强阳性表达,较其他各组增强明显。以上结果表明中、低剂量人脐带间充质干细胞肌肉注射可使扩张型心肌病大鼠血清肝细胞生长因子、白血病抑制因子、粒细胞集落刺激生物因子及血管内皮生长因子水平增高,并增加心肌组织胰岛素样生长因子1、肝细胞生长因子、血管内皮生长因子表达。

BACKGROUND：Studies have shown that intramuscular transplantation of xenogeneic umbilical cord mesenchymal stem cel s in a certain dose range is safe and reliable, and it also confirm that this approach is equal y safe and effective for heart failure in rats with dilated cardiomyopathy. OBJECTIVE：To explore the effect of human umbilical cord mesenchymal stem cel s through intramuscular injection on the cytokine expression in adriamycin-induced dilated cardiomyopathy （DCM） rats. METHODS：Total y 160 rats were randomly divided into control group （n=20） and DCM group （n=140）. Rats in the DCM group were administered adriamycin intraperitoneal y to establish DCM model. The DCM rats were randomly subdivided into model control group （served as model group）, cel supernatant group, the low-dose mesenchymal stem cel group （served as low-dose group）, the middle-dose mesenchymal stem cel group （served as middle-dose group）, and the high-dose mesenchymal stem cel s group （served as high-dose group）. Secondary injection was performed at 4 weeks after first injection. RESULTS AND CONCLUSION：The ELISA test showed that the serum levels of vascular endothelial growth factor （VEGF）, hepatocyte growth factor （HGF）, leukemia inhibitor factor （LIF） and granulocyte macrophage colony stimulating factor （GM-CSF） were higher in the model group than the control group before and after intramuscular injection （P〈0.05）. After intramuscular injection, the levels of HGF, LIF, GM-CSF and VEGF in the low-dose group were increased significantly （P〈0.05）, which were significantly higher than those in the model group （P〈0.05）. The level of LIF in the middle-dose group was significantly elevated after injection （P〈0.05）, while there were no significant differences in HGF, VEGF and GM-CSF levels in the high-dose group before and after intramuscular injection （P〉0.05）. Both the immunohistochemical and RT-PCR results showed that the expressions of insulin-like growth factor-1, VEGF and HGF were increased in al the DCM rats as compared with the control group, which were increased most in the middle-dose group. These findings indicate that low-dose and middle-dose human umbilical cord mesenchymal stem cel s intramuscular injection can increase the serum levels of HGF, LIF, GM-CSF, VEGF and the expressions of IGF-1, HGF and VEGF in the myocardium of DCM rats.