银杏蜜环口服溶液对化学光栓塞型局灶性脑缺血小鼠模型微血管生成与重构的影响  

Effect of Yinxing Mihuan Oral Solution on Neovascularization and Vascular Remodeling in Chemical Photothrombosis-induced Focal Cerebral Ischemia Mouse Model

作　　者：姚明江[1] 张伟[1] 杨斌[1] 宋文婷[1] 王光蕊[1] 王益民 王勇 赫少清 刘建勋[1] YAO Ming-jiang;ZHANG Wei;YANG Bin;SONG Wen-ting;WANG Guang-rui;WANG Yi-min;WANG Yong;HE Shao-qing;LIU Jian-xun(Institute of Basic Medical Sciences of Xiyuan Hospital,Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Beijing 100091,China;Xi'an Buchang Traditional Chinese Cardio-Cerebrovascular Disease Hospital,Xi’an 710082,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,中药药理北京市重点实验室,北京100091 [2]西安步长中医心脑病医院,西安710082

出　　处：《中国实验方剂学杂志》2022年第23期38-43,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国中医科学院科技创新工程重大攻关项目(C12021A01305):中医药治疗缺血性脑卒中“拟临床”药效学评价体系的构建项目;国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202007):中医药防治缺血性心脑血管病传承创新团队项目。

摘　　要：目的:观察银杏蜜环口服溶液对化学光栓塞型局灶性脑缺血小鼠模型的影响,并探讨其在微血管生成与重构方面的作用机制。方法:将50只SPF级雄性C57BL/6J小鼠,随机分为假手术组、模型组、金纳多组(12.5 mg·kg^(-1))、银杏蜜环口服溶液(银蜜)低、高剂量组(412、824 mg·kg^(-1)),每组10只。采用化学光栓阻塞微血管法建立小鼠局灶性脑梗死模型,术后连续灌胃给药14 d。采用改良神经功能缺损评分(mNSS)及前肢抓力测定评价小鼠神经功能;异硫氰酸荧光素葡聚糖(FITCdextran)染色法观察缺血灶周围区域毛细血管密度;苏木素-伊红(HE)染色观察缺血灶周围区域组织病理形态变化;蛋白免疫印迹法(Western blot)检测小鼠缺血灶周边脑组织微血管生成与重构相关指标血管内皮生长因子(VEGF)、血小板-内皮细胞黏附分子(CD31)、缺氧诱导因子-1α(HIF-1α)、血管性血友病因子(vWF)、血管生成素(ANG)蛋白表达。结果:与假手术组比较,模型组小鼠存在显著神经功能缺损(P<0.01),前肢抓力显著下降(P<0.01),缺血灶周围区域毛细血管密度显著增加(P<0.01),存在明显神经元坏死、胶质细胞增生等病理形态改变;给药后14 d,与模型组比较,银蜜高剂量组小鼠神经功能缺损情况显著改善(P<0.01),银蜜低、高剂量组小鼠抓力均显著提升(P<0.01),银蜜高剂量组小鼠缺血灶周围区域毛细血管密度进一步增加(P<0.05),脑组织病理性损伤明显减轻,CD31、ANG、HIF-1α、vWF蛋白表达不同程度上调(P<0.05,P<0.01)。结论:银杏蜜环口服溶液可改善化学光栓塞型局灶性脑缺血小鼠模型运动功能及病理形态,其作用机制可能与促进缺血灶周围区域微血管生成与重构有关。Objective:To investigate the effects and underlying mechanism of Yinxing Mihuan oral solution(YM) on neovascularization and vascular remodeling in chemical photothrombosis-induced focal cerebral ischemia model in mice.Method:Fifty SPF C57BL/6J mice were randomly divided into sham group,model group,ginaton group(12.5 mg·kg^(-1)),and low-(YM-L,412 mg·kg^(-1))and high-dose(YM-H,824 mg·kg^(-1))YM groups,with 10 mice in each group.The focal cerebral ischemia model was established by chemical photothrombosis method.Drugs in each group were administered by gavage for 14 consecutive days after operation.The modified neurological severity score(mNSS)and measurement of forelimb grasping were used to evaluate the neurologic impairment of mice.The vascular density of infarct border-zone(IBZ)was measured by fluorescein labelled dextran(FITC-dextran)method.The morphology of IBZ was evaluated and observed by hematoxylin-eosin(HE) staining.The expression of proteins related to neovascularization and vascular remodeling in brain tissues,including vascular endothelial growth factor(VEGF),platelet and endothelial cell adhesion molecule 1(CD31),hypoxia-inducible factor-1α(HIF-1α),von Willebrand factor(vWF),and angiogenin(ANG),was detected by Western blot.Result:Compared with the sham group,the model group showed manifest neurological deficits(P<0.01),weakened forelimb grasping(P<0.01),increased vascular density of IBZ(P<0.01),and obvious pathological changes,such as neuronal necrosis and gliocyte proliferation.After treatment for 14 days,compared with the model group,the YM-H group showed improved neurological deficits(P<0.01),and the YM-L group and the YM-H group showed strengthened forelimb grasping(P<0.01).Moreover,the YM-L group displayed increased vascular density of IBZ(P<0.05),reduced pathological damage,and up-regulated protein expression of CD31,ANG,HIF-1α,and vWF(P<0.05,P<0.01).Conclusion:YM could improve motor function and pathological morphological impairment in chemical photothrombosis-induced focal cerebral ische

关 键 词：银杏蜜环口服溶液 化学光栓塞 局灶性脑缺血 微血管生成与重构 

分 类 号：R2-0[医药卫生—中医学] R33R289R743.31