| Objective: To explore the efficacy and mechanism of Butylphthalide on ischemicstroke, effect of different doses of Butylphthalide (NBP) on the neurological function,circulating endothelial progenitor cells (EPCs) and neurogenesis in the ischemia regionwere observed in middle cerebral artery occlusion (MCAO)/reperfusion rat.Method:48male Sprague-Dawley rats weighted (250±20)g were selected in thisstudy, which were randomly divided into four groups, ie.sham group, model control group,low dose NBP group and high dose NBP group, and12animals in each group. Neckvessels were isolated but thread embolism was not inserted in sham group. On the otherthree groups, thread embolism method was used to prepare middle cerebral arteryocclusion (MCAO)/reperfusion animal model. Two rats in both sham group and modelcontrol group were performed TTC staining to observe morphological changes in ischemicarea. The rest rats were respectively gavaged with two doses of NBP or control placebo(soybean oil)(2ml/d) for5days. Neurological function of all rats were assessed every day.Peripheral blood was collected before and after treatment to investigate the level ofcirculating EPCs with flow cytometry. Immunochemical staining of CD31and vWF(markers of vascular endothelial cells) was performed to observe angiogenesis after5daysof treatment. All the data were analyzed by GraphPad5.0software analysis system.Results：1）Hemisphere on the side of carotid artery inserted with thread embolism displayedpale color and hemisphere on the other side displayed red color.2）On day1after treatment, neurological function score reduced in low dose NBPgroup (P<0.05), and it significantly reduced in high dose group (P<0.01). The score wassignificantly decreased on day3after treatment in model control group.3）On day1after treatment, neurological function score in low dose NBP group were lower than the control group (P<0.05), the score in high dose group was significantly lowerthan the control group (P<0.01); On day2-5after treatment, the score of different doseNBP groups were significantly lower than the control group (P<0.01);4）On day3after treatment, neurological function score of high dose NBP group wassignificantly lower than the low dose group (P<0.01);5）Compared with ischemia2h, the number of circulating EPCs in sham-operatedgroup rat had no significant change on day5after treatment(P=0.205), other three groupsall increased significantly (all P<0.01);6）On day5after treatment, the number of circulating EPCs in different dose NBPtreatment groups rats were significantly higher than the control group(P<0.01); the numberof circulating EPCs in high dose NBP treatment group was significantly higher than thelow dose group (P<0.01);7）Under the microscope,sham-operated group brain slices anti-CD31-positive cellsand anti-vWF-positive cells were within the normal brain tissue vascular staining.While incontrol group brain slices almost no anti-CD31-positive cells or anti-vWF-positive cellswere visible,suggested vascular damage in cerebral ischemia. In NBP treatment groupsbrain slices,the population of anti-CD31-positive cells and anti-vWF-positive cells werehighly expression, wherein the expression of positive cells were more significant in highdose group.Conclusions:1) MCAO/reperfusion injury can facilitate the mobilization of EPCs;2) NBP can be further mobilized the circulating EPCs, effectively promoteangiogenesis in ischemia areas, improve score of neurological function;the effect ofhigh-dose was more significant than low-dose... |
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