补阳还五汤联合骨髓间充质干细胞移植对大脑中动脉栓塞模型整合素相关因子的影响

发布时间：2018-11-22 07:51

【摘要】：背景:前期研究发现,补阳还五汤联合骨髓间充质干细胞移植可发挥协同抗脑缺血损伤作用。目的:分析骨髓间充质干细胞移植联合补阳还五汤促进脑缺血血管新生的作用。方法:将96只SD大鼠随机分为4组,均制作大脑中动脉栓塞模型。联合组于造模前3 d灌胃给予补阳还五汤10 mL/kg,1次/d,造模成功后2 h灌胃1次,以后每12 h灌胃1次;局灶脑缺血2 h再灌注2 h后,干细胞组、联合组向脑室中注入骨髓间充质干细胞悬液,30 min后再注射抗CD34、CD45抗体;抗体组注射抗CD34、CD45抗体;模型组注射生理盐水。联合组于再灌注后12,24,36,48 h进行血清αVβ3水平及脑组织免疫组织化学、Q-PCR、免疫印迹检测,其余3组均于再灌注后36 h检测上述指标。结果与结论:(1)血清αVβ3水平:抗体组血清αVβ3水平低于模型组(P0.05),干细胞组、联合组高于抗体组(P0.05),且联合组再灌注不同时间点的血清αVβ3水平高于干细胞组(P0.01);(2)免疫组织化学:抗体组CD34阳性细胞数量少于模型组(P0.05),干细胞组、联合组高于抗体组(P0.05);(3)Q-PCR与免疫印迹检测:与模型组相比,抗体组、干细胞组p FAK蛋白表达水平降低(P0.05),FAK基因及蛋白表达变化不大;与其他组比较,联合组FAK蛋白(自再灌注后24 h开始)和p FAK蛋白(自再灌注12 h开始)显著升高(P0.05,P0.01),且随着再灌注时间延长有逐渐升高趋势;(4)结果表明:骨髓间充质干细胞移植联合补阳还五汤可逆转抗CD34+CD45抗体引起的血管内皮细胞数量减少、整合素αVβ3及其下游信号分子表达下降,促进血管新生。
[Abstract]:Background: previous studies have shown that BYHD combined with bone marrow mesenchymal stem cell transplantation can play a synergistic effect on cerebral ischemia injury. Aim: to analyze the effect of bone marrow mesenchymal stem cell transplantation combined with Buyang Huanwu decoction on cerebral ischemic angiogenesis. Methods: 96 SD rats were randomly divided into 4 groups. In the combined group, Buyang Huanwu decoction was given 10 mL/kg,1 / d 3 days before the model was made, and once every 12 hours after the successful establishment of the model. After 2 h of focal cerebral ischemia and 2 h of reperfusion, the stem cells group, combined group, injected bone marrow mesenchymal stem cell suspension into the ventricle, then 30 min later, then injected anti CD34,CD45 antibody; antibody group injected anti CD34,CD45 antibody; model group injected normal saline. The serum 伪 V 尾 3 level, brain tissue immunohistochemistry, Q-PCR and immunoblotting were detected in the combined group at 12: 24, 36 hours after reperfusion, and the above indexes were detected in the other three groups at 36 hours after reperfusion. Results and conclusion: (1) the level of serum 伪 V 尾 3 in the antibody group was lower than that in the model group (P0.05), and the level of 伪 V 尾 3 in the stem cell group was higher than that in the antibody group (P0.05). The serum 伪 V 尾 3 level in the combined group was higher than that in the stem cell group at different time points (P0.01). (2) Immunohistochemistry: the number of CD34 positive cells in the antibody group was less than that in the model group (P0.05), and the number of CD34 positive cells in the stem cell group was higher than that in the antibody group (P0.05). (3) Q-PCR and Western blotting: compared with the model group, the expression of p FAK protein in the antibody group decreased (P0.05) the expression of), FAK gene and protein did not change. Compared with other groups, FAK protein (from 24 h after reperfusion) and p FAK protein (from 12 h after reperfusion) in combined group increased significantly (P0.05 + P0.01), and gradually increased with the prolongation of reperfusion time. (4) the results showed that bone marrow mesenchymal stem cell transplantation combined with Buyang Huanwu decoction could reverse the decrease in the number of vascular endothelial cells induced by anti CD34 CD45 antibody, decrease the expression of integrin 伪 V 尾 3 and its downstream signal molecules, and promote angiogenesis.
【作者单位】：河南中医药大学;河南中医药大学第一附属医院;河南省医药科学研究院;
【基金】：国家自然科学基金课题(81072799/H2708) 河南省高校科技创新团队项目(16IRTSTHN022)~~
【分类号】：R743.3;R-332

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