丹龙醒脑方对脑缺血再灌注大鼠血管新生的影响及机理探讨
发布时间：2018-10-23 18:40
【摘要】：目的：通过动物实验,观察丹龙醒脑方对脑缺血再灌注损伤大鼠大脑血管新生及其因子的影响,探讨其对缺血性中风病理机制的干预作用。 方法：本实验采用MCAO法建立SD雄性大鼠脑缺血再灌注损伤模型,将造模成功的存活大鼠按随机数字表法随机分成6组：丹龙醒脑方大剂量组、丹龙醒脑方中剂量组、丹龙醒脑方小剂量组、尼莫地平阳性对照组、模型组、假手术组,每组8只,除假手术组外,其余各组均采用MCAO法造模,术后2h恢复再灌注,假手术组只结扎颈外动脉,不进行缺血再灌注。术后12h进行神经功能学评分。从术后第一天开始灌胃进行药物干预,每天一次,连续七天,假手术组及模型组给予等体积蒸馏水,七天后麻醉处死,迅速断头取脑,梗死面积测定采用TTC染色法,病例形态学观察采用石蜡包埋切片,在高倍显微镜下观察,通过标记脑缺血组织局部CD34表达来测定内皮细胞数和微血管密度,VEGF、Flk-1、Ang-1、bFGF等生化指标的测定采用免疫组化方法。 结果：(1)神经功能学评分：模型组与假手术组相比,神经功能学评分明显升高(P0.01);各用药组与模型组相比神经功能学评分降低,有统计学意义(P0.05)；丹龙醒脑方各组与尼莫地平组比较无差异(P0.05)；丹龙醒脑方各组间比较无明显差异(p0.05)。(2)脑梗死面积：模型组与假手术组相比,脑梗死面积明显增大(P0.01);各用药组组与模型组相比脑梗死面积降低,有统计学意义(P0.05)；丹龙醒脑方大、中剂量组和尼莫地平组有差异(P0.05),丹龙醒脑方小剂量组与尼莫地平组比较无差异(P0.05)；丹龙醒脑方各组间比较无明显差异(P0.05)。(3)缺血海马区内皮细胞数及微血管密度：模型组与假手术组相比,内皮细胞数和微血管密度均增加(P0.05);各用药组与模型组相比内皮细胞数和微血管密度增加,有统计学意义(P0.05);丹龙醒脑方各组和尼莫地平组无明显差异(P0.05)；丹龙醒脑方各组间比较无明显差异(p0.05)。(4)缺血海马区血管新生因子VEGF、Flk-1、Ang-1、bFGF蛋白表达：模型组与假手术组相比,VEGF、Flk-1、Ang-1、bFGF蛋白表达均增强(P0.05)；各用药组与模型组相比表达增强,有统计学意义(P0.05);丹龙醒脑方各组和尼莫地平组相比无明显差异(P0.05)；丹龙醒脑方各组间比较无明显差异(P0.05)。 结论：丹龙醒脑方能改善脑缺血再灌注后神经功能,缩小梗死面积。脑缺血再灌注损伤后,血管新生因子启动,丹龙醒脑方能增加内皮细胞数及微血管密度,增强血管新生因子VEGF、Flk-1、Ang-1、 bFGF的表达,从而能够促进新生血管的生长及侧枝循环的再建立,这可能是其能改善神经功能的作用机制之一。
[Abstract]:Aim: to observe the effects of Danlong Xingnao recipe on cerebral angiogenesis and its factors in rats with cerebral ischemia-reperfusion injury and to explore the effect of Danlong Xingnao recipe on the pathological mechanism of ischemic stroke. Methods: the MCAO method was used to establish the model of cerebral ischemia-reperfusion injury in SD male rats. The surviving rats were randomly divided into 6 groups according to random digital table: large dose group of Danlong Xingnao recipe, middle dose group of Danlong Xingnao prescription, and middle dose group of Danlong Xingnao recipe. Small dose of Danlong Xingnao Fang group, nimodipine positive control group, model group, sham-operation group, 8 rats in each group, except sham-operation group, all the other groups were made model by MCAO method, and the reperfusion was resumed 2 hours after operation, and the external carotid artery was only ligated in the sham-operation group. No ischemia reperfusion. Neurological function score was performed 12 hours after operation. From the first day after operation, drug intervention was performed by intragastric administration, once a day for seven days. Sham-operation group and model group were given distilled water of the same volume. After seven days, the rats were anesthetized and killed, the head was cut off quickly, and the infarct area was measured by TTC staining. The morphology of the patients was observed by paraffin embedded sections, observed under high power microscope, the number of endothelial cells and microvessel density were measured by labeling the local CD34 expression in ischemic brain tissue, and the biochemical indexes such as VEGF,Flk-1,Ang-1,bFGF were determined by immunohistochemical method. Results: (1) the neurological functional score in the model group was significantly higher than that in the sham operation group (P0.01), and the neurological functional score in each medication group was significantly lower than that in the model group (P0.05). There was no difference between Danlong Xingnaofang group and nimodipine group (P0.05), but there was no significant difference between Danlong Xingnao recipe group (p0.05). (2) in cerebral infarction area: model group and sham operation group, The area of cerebral infarction increased significantly (P0.01); compared with the model group, the area of cerebral infarction decreased significantly in each medication group (P0.05); Danlong Xingnao prescription was large, There was significant difference between middle dose group and nimodipine group (P0.05), but there was no difference between small dose group and nimodipine group (P0.05). There was no significant difference in the number of endothelial cells and microvessel density between the three groups (P0.05). (3): compared with the sham operation group, the number of endothelial cells and microvessel density in the ischemic hippocampus in the model group was higher than that in the sham operation group. The number of endothelial cells and microvessel density increased (P0.05); compared with the model group, the number of endothelial cells and microvessel density increased significantly (P0.05); there was no significant difference between Danlong Xingnaofang group and nimodipine group (P0.05). There was no significant difference in the expression of angiogenic factor VEGF,Flk-1,Ang-1,bFGF in ischemic hippocampus between the three groups (p0.05). (4): the expression of VEGF,Flk-1,Ang-1,bFGF protein in model group was higher than that in sham operation group (P0.05); the expression of VEGF,Flk-1,Ang-1,bFGF protein in each medication group was higher than that in model group. There was statistical significance (P0.05); there was no significant difference between the three groups (P0.05); there was no significant difference between the three groups (P0.05). Conclusion: Danlong Xingnao prescription can improve the nerve function and reduce the infarct area after cerebral ischemia and reperfusion. After cerebral ischemia-reperfusion injury, angiogenic factors were activated. Danlong Xingnao Fang could increase the number of endothelial cells and microvessel density, enhance the expression of angiogenic factor VEGF,Flk-1,Ang-1, bFGF, and thus promote the growth of neovascularization and the re-establishment of collateral circulation. This may be one of its mechanisms for improving neural function.
【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5
本文编号：2290143
[Abstract]:Aim: to observe the effects of Danlong Xingnao recipe on cerebral angiogenesis and its factors in rats with cerebral ischemia-reperfusion injury and to explore the effect of Danlong Xingnao recipe on the pathological mechanism of ischemic stroke. Methods: the MCAO method was used to establish the model of cerebral ischemia-reperfusion injury in SD male rats. The surviving rats were randomly divided into 6 groups according to random digital table: large dose group of Danlong Xingnao recipe, middle dose group of Danlong Xingnao prescription, and middle dose group of Danlong Xingnao recipe. Small dose of Danlong Xingnao Fang group, nimodipine positive control group, model group, sham-operation group, 8 rats in each group, except sham-operation group, all the other groups were made model by MCAO method, and the reperfusion was resumed 2 hours after operation, and the external carotid artery was only ligated in the sham-operation group. No ischemia reperfusion. Neurological function score was performed 12 hours after operation. From the first day after operation, drug intervention was performed by intragastric administration, once a day for seven days. Sham-operation group and model group were given distilled water of the same volume. After seven days, the rats were anesthetized and killed, the head was cut off quickly, and the infarct area was measured by TTC staining. The morphology of the patients was observed by paraffin embedded sections, observed under high power microscope, the number of endothelial cells and microvessel density were measured by labeling the local CD34 expression in ischemic brain tissue, and the biochemical indexes such as VEGF,Flk-1,Ang-1,bFGF were determined by immunohistochemical method. Results: (1) the neurological functional score in the model group was significantly higher than that in the sham operation group (P0.01), and the neurological functional score in each medication group was significantly lower than that in the model group (P0.05). There was no difference between Danlong Xingnaofang group and nimodipine group (P0.05), but there was no significant difference between Danlong Xingnao recipe group (p0.05). (2) in cerebral infarction area: model group and sham operation group, The area of cerebral infarction increased significantly (P0.01); compared with the model group, the area of cerebral infarction decreased significantly in each medication group (P0.05); Danlong Xingnao prescription was large, There was significant difference between middle dose group and nimodipine group (P0.05), but there was no difference between small dose group and nimodipine group (P0.05). There was no significant difference in the number of endothelial cells and microvessel density between the three groups (P0.05). (3): compared with the sham operation group, the number of endothelial cells and microvessel density in the ischemic hippocampus in the model group was higher than that in the sham operation group. The number of endothelial cells and microvessel density increased (P0.05); compared with the model group, the number of endothelial cells and microvessel density increased significantly (P0.05); there was no significant difference between Danlong Xingnaofang group and nimodipine group (P0.05). There was no significant difference in the expression of angiogenic factor VEGF,Flk-1,Ang-1,bFGF in ischemic hippocampus between the three groups (p0.05). (4): the expression of VEGF,Flk-1,Ang-1,bFGF protein in model group was higher than that in sham operation group (P0.05); the expression of VEGF,Flk-1,Ang-1,bFGF protein in each medication group was higher than that in model group. There was statistical significance (P0.05); there was no significant difference between the three groups (P0.05); there was no significant difference between the three groups (P0.05). Conclusion: Danlong Xingnao prescription can improve the nerve function and reduce the infarct area after cerebral ischemia and reperfusion. After cerebral ischemia-reperfusion injury, angiogenic factors were activated. Danlong Xingnao Fang could increase the number of endothelial cells and microvessel density, enhance the expression of angiogenic factor VEGF,Flk-1,Ang-1, bFGF, and thus promote the growth of neovascularization and the re-establishment of collateral circulation. This may be one of its mechanisms for improving neural function.
【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5
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