回族药扎里奴思方联合骨髓间充质干细胞移植对脑缺血再灌注大鼠神经元及P-糖蛋白的影响  

Effect of Hui Medicine Zhali Nusi Fang Combined Bone Marrow Mesenchymal Stem Cells Transplantation on Neurons and P-glycoprotein Expression in Rats After Cerebral Ischemia Reperfusion Injury

作　　者：任非非[1] 刘敬霞[1] 朱万平[2] 虎喜成 刘会贤[3] 刘洋[3] 李娟[3] 

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏医科大学基础医学院,银川750004 [3]宁夏医科大学附属回医中医院,宁夏吴忠751100

出　　处：《中国实验方剂学杂志》2015年第8期125-131,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81260569)

摘　　要：目的:观察扎里奴思方联合骨髓间充质干细胞(BMSCs)移植对脑缺血再灌注(MCAO)模型大鼠神经元及P-糖蛋白(P-gp)表达的影响。方法:250只SD大鼠随机分为假手术组、模型组、扎方组(扎里奴思方组)、移植组和联合组(扎里奴思方和移植组),除假手术组10只外,其余各组再分为1,3,7,14 d组,分别为15只。大鼠术前4 d开始ig给药(14.6 g·kg-1·d-1,假手术组、模型组和移植组给予等体积的生理盐水),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,术后24 h,BMSCs悬浮液经颈内动脉移植入脑(2×106个/200μL);移植后1,3,7,14 d取材,观察海马CA1区神经元密度(ND)和脑组织病理损伤、免疫组化法测P-gp表达变化。结果:模型大鼠海马CA1区ND较假手术组明显降低(P<0.01),病理损伤明显(P<0.01,P<0.05),P-gp表达明显增高(P<0.01);与模型组比较,扎方、移植组3,7,14 d及联合各组ND增加(P<0.01,P<0.05),各组病理损伤减轻,以扎方、移植、联合组7,14 d明显(P<0.01,P<0.05),扎方、移植、联合各组P-gp表达降低(P<0.01);与移植组比较,扎方组1 d ND增高(P<0.01),14 d降低(P<0.05),联合各组ND均增高(P<0.01),联合组7,14 d病理损伤减轻(P<0.05),扎方组1,3,7 d P-gp表达降低,以1 d降低明显(P<0.05),14 d P-gp表达增高(P<0.05),联合各组P-gp表达均降低(P<0.01);扎方与联合组比较,联合各组上述指标均改善(P<0.01,P<0.05);同组间比较,均以7 d变化显著,14 d有明显改善(P<0.01)。结论:脑缺血后脑组织海马CA1区神经元密度及组织病理学均出现不同程度损伤;扎里奴思方和BMSCs移植均可不同程度改善脑缺血后脑组织神经元存活数量及状态,以二者联合作用显著,其机制可能与干预P-gp动态表达有关。Objective： To observe the effect of Hui medicine Zhali Nusi Fang （ZLNS） combined bone marrow mesenchymal stem cells （BMSCs） transplantation on neurons and P-glycoprotein （P-gp） expression in rats after cerebral ischemia reperfusion injury （CIRI）. Method： Two hundred and fifty SD rats were randomly divided into the sham-operated group, the model group, the ZLNS group, the BMSCs transplantation group and the ZLNS combined BMSCs transplantation group. The rats except the 10 rats in the sham-operated group were subdivided into day 1, 3, 7 and 14 groups of 15 each. Middle cerebral artery occlusion （MCAO） model was duplicated by inserting a nylon thread. BMSCs were cultured and amplified by a whole bone marrow adherence method. Drugs were given to the rats by intragastric administration （14.6 g· kg^-1· d^-1）, BMSCs suspension solution were transplanted into brain through carotid artery （2 × 10^6/200 μL）. Rat brain was taken out at 1, 3, 7, 14 days after transplantation. The neuronal density （ND） in hippocampal CA1 area and brain pathological damage （BPD） were observed and determined. The expression of P-gp was measured by using an immunohistochemical method. Result： Compared with the sham-operated group, ND decreased （P 〈 0.01 ）, BPD increased （P 〈 0.01, P 〈 0.05 ）, and the P-gp expression increased significantly （P 〈 0.01 ） in the model group. Compared with the model group, the ND increased at day 3, 7 and 14 in the ZLNS, transplantation and combination groups （P 〈 0.01, P 〈 0.05 ） , the P-gp expression decreased significantly in the ZLNS, transplantation and combination groups （P 〈 0.01 ） , the BPD decreased in all groups, and the results were obvious at day 7 and 14 （P 〈0.01, P 〈0.05）. Compared with the transplantation group, the ND increased at day 1 in ZLNS group （P 〈 0.01 ） , decreased at day 14 in ZLNS group, and increased in combination groups （P 〈0.01 ） , the BPD alleviated at day 7, 14 in the combination group

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 P-糖蛋白 

分 类 号：R285.5[医药卫生—中药学]