脑心一号胶囊联合血管内皮生长因子基因转染骨髓间充质干细胞对脑缺血组织血管生成的影响

Combinatorial effects of naomai yihao capsules and vascular endothelial growth factor gene-transfected bone marrow mesenchymal stem cells on angiogenesis in cerebral ischemic tissues in rats.

机构信息

Guangdong Provincial Hospital, Traditional Chinese Medicine, Guangdong 510120, China.

出版信息

J Tradit Chin Med. 2012 Mar;32(1):87-92. doi: 10.1016/s0254-6272(12)60038-7.

DOI:10.1016/s0254-6272(12)60038-7

PMID:22594109

Abstract

OBJECTIVE

To investigate the combinatorial effects of Naomai Yihao (NMYH) Capsules and vascular endothelial growth factor (VEGF) gene-transfected bone marrow mesenchymal stem cells (BMSCs) on angiogenesis in cerebral ischemic tissues in rats and the mechanism.

METHOD

BMSCs were isolated and cultured from bone marrow by an adherence method. Then, BMSCs were transfected with the eukaryotic expression plasmid pEGFP-VEGF165 by positive ionic liposome transfection. A rat model of middle cerebral artery occlusion (MCAO) was established. Rats were allocated to six groups: model, BMSC, VEGF gene-transfected BMSC transplantation (BMSC/VEGF), NMYH, combined NMYH and BMSC/VEGF (combined treatment group) and sham operation groups. The behavioral rating score (BRS) of rats and the expression of CD34 and VEGF in brain tissue were measured by immunohistochemistry on days 7, 14 and 21 after reperfusion. Angiogenesis was observed and evaluated with laser scanning confocal microscopy.

RESULTS

The BRS of rats in NMYH, BMSC transplantation and combined treatment groups was significantly lower than that of the model group (P < 0.001), with no significant difference between NMYH and transplantation groups (P = 0.619). The expression of CD34 and VEGF in NMYH, transplantation and combined treatment groups increased (P < 0.001), with a significant difference between NMYH and transplantation groups (P < 0.001). The blood vessel area in NMYH, transplantation and combined treatment groups was significantly increased (P < 0.05), without a significant difference between NMYH and transplantation groups (P = 0.873).

CONCLUSIONS

VEGF gene-transfected BMSCs improve angiogenesis in the cerebral ischemic area. NMYH Capsules promote angiogenesis in MCAO rats treated with BMSC transplantation, which show an improved BRS. The mechanism of angiogenesis may be related to up-regulation of VEGF expression.

摘要

目的

探讨脑心一号（NMYH）胶囊联合血管内皮生长因子（VEGF）基因转染骨髓间充质干细胞（BMSCs）对脑缺血组织血管生成的影响及作用机制。

方法

采用贴壁法分离培养大鼠骨髓间充质干细胞，脂质体转染法将真核表达质粒 pEGFP-VEGF165 转染至 BMSCs，建立大脑中动脉闭塞（MCAO）大鼠模型，随机分为模型组、BMSCs 组、VEGF 基因转染 BMSCs 移植组（BMSC/VEGF 组）、NMYH 组、NMYH 联合 BMSC/VEGF 组（联合治疗组）和假手术组。于再灌注后第 7、14、21 天观察各组大鼠行为学评分（BRS），免疫组织化学法检测脑组织 CD34、VEGF 表达，激光扫描共聚焦显微镜观察并评价血管生成情况。

结果

与模型组比较，NMYH 组、BMSC 移植组及联合治疗组大鼠 BRS 降低（P<0.001），NMYH 组与 BMSC 移植组间差异无统计学意义（P=0.619）；NMYH 组、BMSC 移植组及联合治疗组大鼠 CD34、VEGF 表达均增加（P<0.001），NMYH 组与 BMSC 移植组间差异有统计学意义（P<0.001）；NMYH 组、BMSC 移植组及联合治疗组大鼠血管面积增加（P<0.05），NMYH 组与 BMSC 移植组间差异无统计学意义（P=0.873）。