骨髓间充质干细胞脑室内移植联合银杏内酯K腹腔注射治疗大鼠脑缺血再灌损伤  

Bone marrow mesenchymal stem cells intro-ventricular transplantation combined with intraperitoneal injection of ginkgolide K for focal cerebral ischemia/reperfusion injury in rats

作　　者：郝怀勇 贺敏敏 李永涛[1] 吴超[1] 马洪鑫 魏增华[1] 黄保胜 Hao Huaiyong;He Minmin;Li Yongtao;Wu Chao;Ma Hongxin;Wei Zenghua;Huang Baosheng(Department of Neurosurgery, Tengzhou Central People’s Hospital Affiliated to Jining Medical College, Tengzhou 277599, China;Department of Paediatrics, Tengzhou Central People’s Hospital Affiliated to Jining Medical College, Tengzhou 277599, China;Department of Neurosurgery, Sir Run Run Hospital, Nanjing Medical University, Nanjing 210006, China)

机构地区：[1]济宁医学院附属滕州市中心人民医院神经外科,山东滕州277599 [2]济宁医学院附属滕州市中心人民医院儿科,山东滕州277599 [3]南京医科大学附属逸夫医院神经外科,210006

出　　处：《中华实验外科杂志》2019年第8期1377-1380,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金项目(81501059);江苏省“科教强卫工程”青年医学人才项目(QNRC2016858);江苏省自然科学基金青年基金项目(BK20171064);滕州市科技发展计划(201510012).

摘　　要：目的观察大鼠骨髓间充质干细胞(BMSCs)脑室内移植联合银杏内酯K(GK)腹腔注射对大鼠局灶性脑缺血/再灌注损伤的神经保护作用.方法体外培养BMSCs并诱导向神经细胞样分化.制备大鼠大脑中动脉栓塞脑缺血再灌注(MCAO)模型.实验动物按事后机率均等单盲随机分为5组,分别设为Sham组,生理盐水腹腔注射组、GK腹腔注射组、BMSCs移植+生理盐水腹腔注射组、BMSCs移植+GK腹腔注射组.于脑缺血再灌注后1h腹腔给药,于24 h后移植BMSCs,于72、120、168 h进行神经功能测定;然后分别处死动物,取脑染色,观察脑梗死面积,测定脑损伤程度,脑水肿程度等.蛋白质印迹法(Western blot)检测各组脑组织中血管内皮生长因子(VEGF)蛋白含量,以评价GK是否通过增加脑组织中VEGF的含量从而提高脑缺血灶处血管再生.结果 GK可以明显增加大鼠脑缺血脑组织内VEGF的蛋白含量.BMSCs脑室内移植联合GK腹腔注射组中大鼠脑缺血区细胞凋亡数[(18.8±3.6)个/高倍镜视野,F=167.531,P<0.01]、脑梗死体积[(17.31±5.24)%,F=34.910,P<0.01]及脑含水量[(73.55±0.53)%,F=65.128,P<0.01]较其他各组明显降低,治疗后大鼠神经缺陷症状评分[120 h:(3.51 ±0.46)分,F=15.473,P<0.01;168 h:(2.03 ±0.35)分,F=21.711,P<0.01]较对照组下降明显.结论 BMSCs脑室内移植联合GK腹腔注射对大鼠脑缺血再灌注损伤有明显协同神经保护作用.GK的作用是通过增加脑缺血灶VEGF的含量来实现的.Objective To investigate the neuroprotective effect of combined application of bone marrow mesenchymal stem cells (BMSCs) and Ginkgolide K (GK) intraperitoneal injection for rat focal cerebral ischemia. Methods In all rats, the middle cerebral artery occlusion (MCAO) was produced by occlusion of right internal carotid artery with a nylon monofilament. Fifty male Sprague-Dawley (SD) rats were divided into five groups by the random digits table, 10 rats in each group, including Sham group, saline solution intraperitoneal injection group, GK intraperitoneal injection group, BMSCs+ saline solution intraperitoneal group, and BMSCs+ GK intraperitoneal injection group. The neurological defective scores were assessed at 72, 120, 168 h after reperfusion respectively in each experiment and all the animals were then decapitated to determine the brain infarct volume and brain water content after 168 h. The expression of vascular endothelial growth factor (VEGF) was detected by Western blotting. Results The expression of VEGF was increased significantly after intraperitoneal injection of GK. Combined treatment of BMSCs and GK could significantly decrease the number of apoptosis cells [(18.8±3.6)/HP, F=167.531, P<0.01] and infarct volumes [(17.31±5.24)%, F=34.910, P<0.01], brain water content in ischemia hemisphere [(73.55±0.53)%, F=65.128, P<0.01]. The values of neurological defective scores in BMSCs+ GK injection group were significantly lower than the rest groups [120 h:(3.51±0.46), F=15.473, P<0.01;168 h:(2.03±0.35), F=21.711, P<0.01]. Conclusion Combined treatment of BMSCs intro-ventricular transplantation and GK intraperitoneal injection produces a synergistic neuroprotective effect.

关 键 词：血管内皮生长因子 脑缺血 骨髓间充质干细胞 银杏内酯K 

分 类 号：R743.3[医药卫生—神经病学与精神病学]