人参皂苷Rd通过SHH信号通路促进MCAO模型大鼠血管新生机制研究  

Mechanism of ginsenoside Rd in promotion of angiogenesis by regulaiton of SHH signaling pathway activity in a rat model of middle cerebral artery occlusion(MCAO)

作　　者：王宝祥 许俊杰 WANG Bao-xiang;XU Jun-jie(Department of Neurology,The First Hospital of Jiaxing City,Jiaxing,Zhejiang 314000,China)

机构地区：[1]浙江省嘉兴市第一医院神经内科,嘉兴314000

出　　处：《浙江中西医结合杂志》2023年第4期308-312,共5页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省医药卫生科技计划项目(No.2020RC123)。

摘　　要：目的探讨人参皂苷Rd对大脑中动脉闭塞(MCAO)模型大鼠血管新生的影响及相关机制。方法40只SD大鼠随机数字表法分为假手术组、模型组、人参皂苷Rd组(Rd组)、人参皂苷Rd+环靶明(CYC)组(Rd+CYC组),每组10只。采用线栓塞法建立MCAO模型大鼠,假手术组仅分离左侧颈总动脉、颈外动脉和颈内动脉。造模成功后,Rd组大鼠按照50 mg/kg给予人参皂苷Rd,Rd+CYC组按50μg/kg给予CYC腹腔注射,假手术组和模型组给予同等剂量的生理盐水,每天给药1次,连续给药2周。各组大鼠神经功能评分,TTC方法检测脑梗死体积,显微镜下观察脑组织毛细血管密度,蛋白质免疫印记检测血管生成相关分子和声波刺猬信号蛋白(SHH)、血管内皮生长因子(VEGF)、血管紧张素-1(ANG-1)、血管紧张素-2(ANG-2)蛋白表达水平。结果与模型组比较,Rd组大鼠神经功能评分显著降低[(1.000±0.471)分比(3.100±0.738)分,P<0.01];脑梗死体积显著减少[(18.120±3.735)%比(33.920±2.954)%,P<0.01];脑梗死区域毛细血管数量显著增加[(389.60±23.34)条比(54.37±11.21)条,P<0.01];血管生成相关分子SHH、VEGF、ANG-1、ANG-2蛋白表达水平显著升高[SHH:(1.267±0.038)比(0.533±0.043);VEGF:(0.866±0.035)比(0.596±0.055);ANG-1:(0.880±0.036)比(0.510±0.056);ANG-2:(0.870±0.040)比(0.386±0.045),P均<0.01]。与Rd组比较,Rd+CYC组大鼠神经功能评分显著升高[(2.700±0.675)分比(1.000±0.471)分,P<0.01];脑梗死体积显著增加[(28.910±4.491)%比(18.120±3.735)%,P<0.01];脑梗死区域毛细血管数量显著减少[(180.90±37.37)条比(389.60±23.34)条,P<0.01];血管生成相关分子VEGF、ANG-1、ANG-2蛋白表达水平显著下降[VEGF:(0.600±0.007)比(0.866±0.035);ANG-1:(0.570±0.027)比(0.880±0.036);ANG-2:(0.476±0.038)比(0.87±0.04),P均<0.01]。结论人参皂苷Rd可能通过SHH信号通路促进脑梗死区域血管生成,改善MCAO模型大鼠预后并发挥保护作用。Objective To assess the effects of ginsenoside Rd on regulation of angiogenesis in a rat model of the middle cerebral artery occlusion(MCAO)and then explored the underlying molecular events.Methods Forty Sprague-Dawley rats were randomly divided into sham operation,MCAO model control,model+ginsenoside Rd(Rd),and model+ginseng Saponin Rd+cyclopamine(Rd+CYC)groups(n=10 per group).The model rats were established by thread embolization and their neurological function scores and assessed and the volume of the cerebral infarction(CI)was detected by the TTC method,while the capillary density of rat brain tissues was observed in H&E stained tissue section under a microscope and expression of the angiogenesis-related molecules,sonic hedgehog(SHH),vascular endothelial growth factor(VEGF),angiotensin-1(ANG-1),and angiotensin-2(ANG-2)proteins were analyzed using Western blot.Results The Rd treatment significantly reduced the neurological function score in rats(1.000±0.471 vs.3.100±0.738;P<0.01)and the CI volume(18.120±3.735 vs.33.920±2.954;P<0.01)compared those of the model control rats,whereas the Rd treatment significantly increased the number of capillaries in the CI area(54.37±11.21 vs.389.60±23.34)compared to that of the model control mice(P<0.01).In addition,the ginsenoside Rd treatment significantly increased levels of SHH(1.267±0.038 vs.0.533±0.043),VEGF(0.866±0.035 vs.0.596±0.055),ANG-1(0.88±0.036 vs.0.51±0.056),and ANG-2(0.87±0.04 vs.0.386±0.045)proteins compared with the model controls(all P<0.01).However,the SHH-specific inhibitor CYC was able to weaken the protective and angiogenesis effect of ginsenoside Rd,but decrease levels of VEGF,ANG-1,and ANG-2 proteins in mice.Conclusion Ginsenoside Rd promotes angiogenesis in the CI region through activation of the SHH signaling and prognosis of MCAO model rats in vivo.

关 键 词：大鼠 人参皂苷 脑梗死 大脑中动脉闭塞 声波刺猬信号蛋白 

分 类 号：R285.5[医药卫生—中药学]