VEGF基因修饰MSCs移植对脑梗死大鼠血管源性机制的实验研究

目的观察经尾静脉注射携带VEGF基因的MSCs移植对脑梗死模型大鼠血管生成方面的影响。方法采用直接贴壁全骨髓法分离纯化MSCs,通过脂质体转染技术将pIRES2-EGFP-VEGF导入MSCs,另采用改良Zea Longa线栓法将40只SD大鼠制成左侧大脑中动脉栓塞/再灌注模型,并随机分4组:VEGF-MSCs组、MSCs组、PBS组、Model组;造模24 h后Model组不做处理,前3组大鼠分别经尾静脉注射1 ml VEGF-MSCs、MSCs、PBS悬液;细胞移植7d后用免疫组织化学法测定脑梗死周围Ang-2、CD34的表达水平。结果 VEGF-MSCs组和MSCs组Ang-2、CD34阳性表达水平较PBS组、Model组高(P<0.05),且VEGF-MSCs组较MSCs组更高(P<0.05);Model组与PBS组差异不明显(P>0.05)。结论经尾静脉注射携带VEGF基因的MSCs移植入MCAO模型大鼠可以促进缺血脑组织新生血管的形成,其机制可能为VEGF上调缺血周边区域Ang-2、CD34的表达水平。     

电针结合安脑丸对脑梗死大鼠模型的神经保护作用

目的 在短暂性大脑中动脉栓塞缺血再灌注(transient middle cerebral artery occlusion infarction/reperfusion,tMCAO I/R)模型大鼠中证实电针结合安脑丸治疗对其具有神经保护作用,并对脑梗死后神经保护的机制进行初步探索.方法 SD大鼠随机分为假手术组、模型组、安脑丸组和电针结合安脑丸组.线栓法制作SD大鼠左侧短暂性大脑中动脉栓塞模型.mNSS评分法对各组大鼠各个时间点进行行为学评分.实时荧光定量PCR技术对各组大鼠各个时间点Bcl-2 mRNA转录水平进行分析.免疫组化法观察脑梗死后10 d各组大鼠脑组织Caspase-3阳性细胞表达,Western blot分析各组大鼠脑组织Caspase-3蛋白表达水平.TUNEL原位细胞凋亡检测脑梗死后10 d各组大鼠梗死区边缘神经元细胞凋亡数量.结果 假手术组(正常)mNSS评分为0,未见脑梗死,偶见生理性凋亡细胞.安脑丸组和电针结合安脑丸组在4、7、10 d mNSS评分明显低于模型组(均P<0.05),其中,电针结合安脑丸组更低(P<0.05).实时荧光定量PCR的检测结果 显示脑梗死后Bcl-2 mRNA转录水平升高,安脑丸组和电针结合安脑丸组与模型组相比升高更明显(均P<0.05),其中,电针结合安脑丸组较安脑丸组更高(P<0.05).免疫组化及Western blot的检测结果 都显示,脑梗死后10 d安脑丸组和电针结合安脑丸组大鼠脑组织Caspase-3阳性细胞表达数及蛋白表达水平要明显低于模型组(均P<0.05),其中电针结合安脑丸组较安脑丸组更低(P<0.05).TUNEL检测结果 显示梗死后10 d安脑丸组和电针结合安脑丸组的神经元细胞凋亡数量明显低于模型组(均P<0.05),电针结合安脑丸组较安脑丸组更低(P<0.05).结论 中药安脑丸治疗 tMCAO I/R模型大鼠,可减少大鼠脑梗死区边缘神经元细胞凋亡数量,改善神经功能,结合电针治疗效果更为显著.其神经保护作用机制可能与针药结合治疗后上调了Bcl-2 基因表达,抑制Caspase-3的表达有关.     

急性前循环串联闭塞血管开通治疗临床研究

目的 评价不同血管内介入方法开通不同病因所致急性前循环串联闭塞患者的有效性、安全性及预后.方法 回顾性分析2017年2月至2020年10月在厦门大学附属中山医院接受介入治疗的44例急性前循环串联闭塞性脑梗死住院患者.根据中国缺血性卒中亚分类(CISS)病因分型分为大动脉粥样硬化斑块破裂继发原位血栓形成组(LAA组,n=...     

Over-expression of VEGF in marrow stromal cells promotes angiogenesis in rats with cerebral infarction via the synergistic effects of VEGF and Ang-2

This study explored whether the transplantation of modified marrow stromal cells (MSCs) has angiogenic effects in a left middle cerebral artery occlusion infarction/reperfusion (MCAO I/R) rat model and preliminarily examined the mechanism of angiogenesis following cerebral infarction.MSCs were isolated by using a direct adherent method and cultured.Vascular endothelial growth factor (VEGF) was transfected into MSCs by employing the liposome transfection.The transfection efficiency was measured by the optical density method.The protein expression of VEGF gene before and after transfection was measured by Western blotting.SD rat model of transient occlusion of the left middle cerebral artery was established by using an approach of intra-luminal occlusion.Tetrazolium (TTC) and HE staining were performed to observe the cerebral infarction.ELISAs were used to measure the levels of VEGF in the rat cerebral tissues.The expression patterns of angiopoietin-2 (Ang-2) and CD34 in cells surrounding the area of infarction were immunohistochemistrically oserved.Ang-2 protein expression in the tissue surrounding the area of infarction was measured by Western blotting.VEGF expression in the MSCs increased after transfection at a rate of approximately 28%±3.4%.ELISA showed that the expression of VEGF in the cerebral tissue was significantly increased after induction of infarction,peaking on the 4th day and decreasing to the levels of the sham surgery group (normal) within 7 to 10 days.The VEGF level was significantly higher at each time point in the VEGF-MSC and MSC groups compared to the model group.Moreover,the VEGF level was higher in the VEGF-MSC group than in the MSC group and stayed relatively high until the 10th day.The immunohistochemical results showed that 10 days after the infarction,the number of Ang-2 and CD34-expressing cells in the area surrounding the infarction was significantly higher in the VEGF-MSC group and the MSC group compared to the model group.Moreover,the VEGF level was higher in the VEGF-MSC group th