回医扎里奴思方对脑缺血再灌注损伤大鼠骨髓间充质干细胞移植向神经样细胞分化的影响  

Influence of Hui medicine Zhali Nusi Formula on differentiation of stem cells in bone marrow mesenchyme of rats with cerebral ischemia reperfusion injury

作　　者：刘敬霞[1] 任非非[2] 刘会贤[2] 刘洋[2] 李娟[2] 虎喜成 刘抒雯 

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏医科大学附属回医中医院,宁夏吴忠751100

出　　处：《中华中医药杂志》2016年第7期2567-2573,共7页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:观察扎里奴思方对脑缺血再灌注损伤(CIRI)大鼠骨髓间充质干细胞(BMSCs)移植向神经样细胞分化的影响。方法:SD大鼠随机分为假手术组、模型组、扎方组、移植组和联合组;线栓法制备大鼠MCAO模型,体外全骨髓贴壁筛选法培养及扩增BMSCs并于移植前48h用Brdu标记;大鼠灌胃给药(1.46g·100g^(-1)·d^(-1)),BMSCs悬浮液经颈内动脉移植入脑(2×106/200μL);移植后1、3、7、14d取材,免疫荧光双标检测Brdu、Brdu/NSE、Brdu/GFAP表达,Western blot检测NGF、GDNF蛋白表达。结果:模型各组NSE、GFAP、GDNF表达较假手术组增高(P<0.01),与本组1d比较,3、14d NGF表达升高(P<0.01);与模型组同期比较,扎方组3、7d NSE表达减低(P<0.01),14d增高(P<0.01),扎方组1、7、14d GFAP和GDNF表达增高(P<0.01),各组NGF表达增高(P<0.01),移植各组NSE、NGF、GDNF表达增高(P<0.01,P<0.05),3、7、14d组GFAP表达增高(P<0.01),联合各组以上指标均增高(P<0.01),移植及联合各组见Brdu、Brdu/NSE、Brdu/GFAP表达;与移植组比较,扎方组3、7、14d NSE和GFAP表达减低(P<0.01),扎方组1、3d NGF减低(P<0.01),各组GDNF减低(P<0.01),联合各组NSE和GFAP表达增高(P<0.01),Brdu、Brdu/NSE、Brdu/GFAP增多,各组NGF和GDNF表达增高,以1、14d组明显(P<0.01,P<0.05);扎方与联合组比较,联合各组以上指标均增高(P<0.01);同组间比较,各组NSE、GFAP和NGF均随时间增高,GFAP 7d达高峰,NGF 3d达高峰(P<0.01),模型和联合各组GDNF呈先增后减趋势,3d达高峰(P<0.01),扎方和移植各组呈递减趋势(P<0.01)。结论:扎里奴思方可显著促进CIRI后BMSCs脑内移植并向神经样细胞分化,促进损伤后神经再生修复,其机制可能与干预脑内NGF和GDNF的动态表达有关。Objective： To observe the influence of Zhali Nusi Formula（ZLNSF） on differentiation of bone marrow mesenchyme stem cells（BMSCs） of cerebral ischemia reperfusion injury（CIRI） rat models. Methods： SD rats were randomly divided into sham-operation group, model group, ZLNSF group, transplantation group, and integration group. MCAO rat models were established by using suture-occluded method. The BMSCs were cultured and proliferated by using adherence screening method in vitro, and BMSCs were marked with Brdu before 48 h of transplantation. Rats in each group received gavage administration with corresponding drugs（1.46g·100g^-1·d^-1）, and BMSCs suspension liquid（2×106/200μL） was transplanted into the brain via internal carotid. The rats were sacrificed and the tissues were obtained on the 1st, 3rd, 7th, 14 th day after transplantation. The expression of Brdu, Brdu/NSE, and Brdu/GFAP was tested by using double-labelling immunofluorescence, and the protein expression of NGF and GDNF was tested by using Western blot. Results： The expression of NSE, GFAP, GDNF, and NGF of rats in model group on the 3rd and 14 th day was higher than those of sham-operation group（P0.01）. Compared with model group, the expression of NSE of rats in ZLNSF group on the 3rd and 7th day decreased（P0.01）, while on the 14 th day the NSE expressionincreased（P0.01）, and the expression of GFAP and GDNF of rats in ZLNSF group on the 1st, 7th, 14 th day was increased（P0.01）. The expression of NGF of rats in all the groups and the expression of NSE, NGF, GDNF of rats in transplantation groups were all increased（P0.01, P0.05）. The expression of GFAP in the 3rd, 7th, 14 th groups increased significantly（P0.01）, and the above indexes of rats in all the integration groups were all increased significantly（P0.01）. The expression of Brdu, Brdu/NSE, and Brdu/GFAP was all significant in transplantation and integration groups. Compared with transplantation group, the expression of NSE and GFAP of rats in ZLN

关 键 词：脑缺血再灌注 扎里奴思方 骨髓间充质干细胞 神经元特异性烯醇化酶 胶质纤维酸性蛋白 神经营养因子 

分 类 号：R29[医药卫生—民族医学]