摘要
OBJECTIVE:To investigate the combinatorial effects of Naomai Yihao(NMYH) Capsules(脑脉一号胶囊) 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-VEGF 165 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 rat and the expression of CD34 and VEGF in brain tis sue were measured by immunohistochemistry on days 7,14 and 21 after reperfusion.Angiogenesi was observed and evaluated with laser scanning confocal microscopy.RESULTS:The BRS of rats in NMYH,BMSC transplan tation 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 andVEGF in NMYH,transplanta tion 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 com bined 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 im prove 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 angio genesis may be related to up-regulation ofVEGF ex pression.
OBJECTIVE：To investigate the combinatorial effects of Naomai Yihao（NMYH） Capsules（脑脉一号 胶 囊） 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-VEGF 165 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 rat and the expression of CD34 and VEGF in brain tis sue were measured by immunohistochemistry on days 7,14 and 21 after reperfusion.Angiogenesi was observed and evaluated with laser scanning confocal microscopy.RESULTS：The BRS of rats in NMYH,BMSC transplan tation 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 andVEGF in NMYH,transplanta tion and combined treatment groups increased（P 0.001）,with a significant difference between NMYH and transplantation groups（P0.001）.The blood vessel area in NMYH,transplantation and com bined treatment groups was significantly increased（P0.05）,without a significant difference between NMYH and transplantation groups（P=0.873）.CONCLUSIONS：VEGF gene-transfected BMSCs im prove 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 angio genesis may be related to up-regulation ofVEGF ex pression.
基金
Supported by the Research Fund for the Doctoral Program of Higher Education of China (No.20070572004,20104425120009)
Guangdong Natural Science Fund(No.06301402)
Traditional Chinese Medicine Master Education Program of Tongji University (Sponsored by State Ad-ministration of Traditional Chinese Medicine of the People'sRepublic of China,No:[2008]185)
