【摘要】 目的:探讨通心络胶囊对大鼠脑梗死皮层血管新生与神经元凋亡的影响。方法:M C A o大鼠分成通心络干预组与对照组,于梗死后第4天处死大鼠,免疫组化检测脑缺血皮质V E G F、V III因子及凋亡蛋白酶caspase-3表达,W estern blot半定量检测V E G F、caspase-3蛋白表达情况,TU N E L法检测梗死皮层神经元凋亡。结果:通心络干预组缺血皮质血管密度和V E G F阳性细胞数量均较对照组为高(P均<0.01)。W estern blot显示脑缺血皮层同侧V E G F121、V E G F165有明显表达,而缺血对侧表达很少;与对照组相比,通心络组缺血侧V E G F121、V E G F165蛋白表达均较对照组显著增高(P<0.01)。caspase-3免疫组化显示通心络组缺血侧阳性细胞数量较对照组为低;两组大鼠缺血侧与对侧皮层W estern blot均可显示出pre-caspase-3和活化的caspase-3两条带,以活化的caspase-3分析,对照组缺血侧caspase-3含量明显增高,而通心络组caspase-3含量明显降低,与对侧等其他各组间没有明显差异;TU N E L进一步显示通心络组缺血侧皮质细胞凋亡明显低于对照组(P<0.05)。结论:通心络胶囊通过增加V E G F表达而诱导脑梗死大鼠皮层血管增生,同时阻滞caspase-3活化而减轻神经元凋亡。更多还原

【Abstract】 Objective To explore the effect of Tongxinluo capsules on cortex angiogenesis and neuron apoptosis within focal cerebral ischemic rat brain. Methods Wistar rats were divided in two groups:control group and Tongxinluo capsule group and both subjected to MCAo surgery model. The immunohistochemistry was used to determine the VEGF,VIII factors and caspase-3 positive cells, Western blot to evaluate the expression level of VEGF and caspase-3 protein and TUNEL to quantify the apoptosis neuron cells in ischemic cortex. Results Compared with the control groups,Tongxinluo capsules significantly increased the vascular density and VEGF positive cell numbers(P< 0.01). Western blots demonstrated prominence of two isoforms of VEGF protein, VEGF121 and VEGF165, in ipsilateral ischemic cortex but scarcity in contralateral cortex in both groups. Semi-quality analysis demonstrated that both levels of VEGF121 and VEGF165 were much higher in Tongxinluo groups than those in control groups(P< 0.01). Fewer caspase-3 positive cells within ischemic cortex in Tongxinluo group were counted than those in the control group(P< 0.01).In two groups, caspase-3 Western blot detected two bands corresponding to Pre-caspase-3 and activated caspase-3 within both ischemic ipsilateral and contrateral cortex. The untreated groups obtained the most expression of activated caspase-3 within ischemic cortex among all subgroups(P< 0.01), but the expression of activated caspase-3 in Tongxinluo group failed to achieve the statistical significance either in its own control or compared with ischemic contralateral cortex of control groups(P >0.05). Moreover, the cortex TUNEL-positive cells in Tongxinluo group was much fewer than those in the control group(P< 0.05). Conclusions Tongxinluo capsule can enhance angiogenesis within ischemic cortex by up-regulating the expression of VEGF protein, while can decrease neuron apoptosis, induced by ischemia and reperfusion injury, through preventing caspase-3 from being activated.更多还原