心肌1号对大鼠异丙肾上腺素心肌损伤的骨髓干细胞修复作用研究

目的 探讨临床治疗扩张型心肌病的中药经验复方心肌1号对大鼠异丙肾上腺素心肌损伤的骨髓干细胞修复作用.方法 采用Wister大鼠30只,随机分为3组,健康对照组10只,每天同一时间皮下注射生理盐水6 ml,连续18 d;异丙肾上腺素模型组10只,连续3 d腹部皮下注射异丙肾上腺素15 mg/kg,后每天同一时间皮下注射生理盐水6 ml,连续15 d;心肌1号干预组10只,连续3 d腹部皮下注射异丙肾上腺素15 mg/kg,后予心肌1号20.4 g·kg-1·d-1灌胃,连续15 d.3组大鼠在第4天尾静脉取血1 ml,心肌1号干预组在心肌1号灌胃第12天再次尾静脉取血1 ml,分别加入CD34和CD44单克隆抗体,用流式细胞仪检测外周血CD34和CD44细胞占单个核细胞的比例.同时3组大鼠均在取血完成后腹腔注射Brdu(50 mg/kg)2次,间隔2 h.第18天之后再观察2周,在第32天用戊巴比妥麻醉处死3组大鼠,取出大鼠心脏,分别行HE染色与Brdu免疫组化染色,在Brdu染色阳性的基础上再行Brdu、TNI免疫组化双标染色,Brdu染色阴性的不再行双标染色.结果 心肌1号干预组外周血CD34和CD44细胞占单个核细胞的比例显著高于健康对照组和异丙模型组及用药前,同时心肌组织免疫组化显示心肌1号干预组有新生细胞生成,并且新生细胞表达心肌特异性蛋白肌钙蛋白I(TnI).结论 (1)心肌1号能提高实验大鼠外周血CD34、CD44细胞占单个核细胞的比例;(2)心肌1号有促进实验大鼠新生细胞生成的作用;(3)新生细胞有心肌特异性蛋白TnI表达.心肌1号可能有动员骨髓干细胞的增殖并释放入外周血中的作用,且动员的骨髓干细胞有归巢至受损心肌并定向分化为心肌细胞可能.

Effects of mobilization of bone marrow stem cells by traditional chinese medicine on regeneration of myocardial cells after myocardial injury induced by isoproterenol in rats

Objective To study the myocardium regenerative potential of bone marrow stem cells mobilized by traditional chinese medicine after myocardial injury in rats,which is very effective in treatment of patients with dilated cardiomyopathy in clinical.Methods Thirty Wistar rats were randomly divided into 3 groups：（1）placebo group（n=10）：each rat was injected normal saline through subcutaneous way for 18 days.（2）isoproterenol group（n=10）：15 mg·kg-1·d-1 isoproterenol was injected through subcutaneous way for 3 days,then inject normal saline for 15 days.（3）traditional chinese medicine group：15 mg·kg-1·d-1 isoproterenol was injected through subcutaneous way for 3 days,then traditional chinese medicine（20.4 g·kg-1·d-1）was fed for 15 days.Blood was taken from tail vein for each group to measure the proportion of CD34 and CD44 cells in mononuclear cells in peripheral blood.All rats were killed after 32 days,and the hearts were taken out to make HE stain and Brdu immunohistochemical stain.Results The comparison of proportion of CD34 and CD44 cells in mononuclear cells in peripheral blood between post and pre-traditional chinese medicine group,placebo group,isoproterenol group had statistical difference（P0.01）.Myocardium immunohistochemical stain showed many Brdu stained cells in traditional chinese medicine group,and the Brdu stained cells showed TnI expression.Conclusions Traditional chinese medicine can improve the proportion of CD34 and CD44 cells in mononuclear cells in peripheral blood,promote the generation of new cells,and the new cells can express TnI（cardiac-specific protein）.