大黄苷元对骨髓间充质干细胞移植脑缺血大鼠基质金属蛋白酶的影响_临床医学论文.docVIP

大黄苷元对骨髓间充质干细胞移植脑缺血大鼠基质金属蛋白酶的影响_临床医学论文 大黄苷元对骨髓间充质干细胞移植脑缺血大鼠基质金属蛋白酶的影响_临床医学论文 作者：李建生, 刘敬霞, 孙捷, 赵跃武, 苏静, 李宁, 郭晓燕 【摘要】 　　目的：观察大黄苷元对脑缺血再灌注损伤大鼠骨髓间充质干细胞(bone marrow stromal cells, BMSCs)移植术后脑组织病理改变及基质金属蛋白酶的影响。方法：体外全骨髓细胞贴壁筛选培养法扩增BMSCs；线栓法制备大脑中动脉阻塞（middle cerebral artery occlusion, MCAO）模型。190只大鼠随机分为假手术组、模型组、大黄苷元组、BMSCs移植组（简称移植组）和BMSCs移植联合大黄苷元组（简称联合组）。再灌注后24 h将BMSCs由同侧颈内动脉移植入移植组和联合组大鼠脑内；大黄苷元组和联合组予大黄苷元灌胃用药。移植后7、14和28 d，模型各组取脑组织用于检测。光镜下观察脑组织微血管病理改变；免疫组织化学法测定BMSCs免疫球蛋白抗体G(immunoglobulin G, IgG)、Ⅳ型胶原（type Ⅳ collagen, colⅣ）、基质金属蛋白酶9（matrix metalloproteinase9, MMP9）和金属蛋白酶组织抑制物1（tissue inhibitor of metalloproteinase）的表达。结果：与正常对照组比较，模型组大鼠微血管残缺、破损明显，脑组织中IgG和MMP表达增强，ColⅣ和TIMP表达减弱。与模型组比较，大黄苷元组、移植组和联合组大鼠微血管结构改善明显；大黄苷元组和联合组各时间点大鼠脑组织中IgG表达和MMP活性减弱，ColⅣ和TIMP表达增强。联合组大鼠于移植7 d时的微血管结构较移植组完整，其各时间点大鼠脑组织中ColⅣ表达较移植组增强，14 d时大鼠脑组织中MMP活性较大黄苷元组降低，TIMP表达增强，28 d时TIMP表达较移植组增强。结论：大黄苷元可使脑缺血再灌注损伤大鼠微血管基底膜ColⅣ降解减少，通透性降低，并与BMSCs有协同作用，其作用机制可能与增加TIMP表达以调节MMP平衡有关。 【关键词】 脑缺血 大鼠 大黄苷元 骨髓间充质干细胞 血脑屏障 基质金属蛋白酶 　　Methods: The BMSCs were purified and amplified by methods of adherence and selection in vitro. One hundred and ninety rats were randomly divided into shamoperated group, untreated group, rhubarb aglycone group, BMSC transplantation group (abbreviated as transplantation group) and BMSCs combined with rhubarb aglycone group (abbreviated as combination group). Middle cerebral artery occlusion (MCAO) model was duplicated with nylon thread. Rats of transplantation and combination group were transplanted with BMSCs via carotid artery after 24hour reperfusion. Rhubarb aglycone was used by intragastric administration in the rhubarb aglycone group and the combination group. The brain samples were taken at 7, 14 and 28 days after transplantation. Brain micrangium pathological changes were observed by light microscope, and immunohistochemical method was used to determine the expressions of immunoglobulin G (IgG), type Ⅳ collagen (Col Ⅳ), matrix metalloproteinase9 (MMP9), and tissue inhibitor of metalloproteinase1 (T