摘要
目的探讨通窍益智颗粒对血管性痴呆大鼠血脑屏障及血管新生的影响及其可能机制。方法选取健康清洁级雄性SD大鼠130只,随机取20只作为假手术组,其余大鼠以双侧颈总动脉永久性结扎法建立血管性痴呆模型。将造模成功大鼠随机分为模型组、尼莫地平组和通窍益智颗粒高、中、低剂量组,每组20只。尼莫地平组给予尼莫地平6.25 mg/kg灌胃,通窍益智颗粒高、中、低剂量组分别给予通窍益智颗粒23.54 g/kg、11.77 g/kg、5.88 g/kg灌胃,假手术组和模型组给予等量生理盐水灌胃。灌胃28 d后,Morris水迷宫评估各组大鼠学习和记忆能力,伊文思蓝评估血脑屏障通透性,透射电镜观察血脑屏障超微结构,免疫荧光标记海马CA1区CD31蛋白观察微血管密度,Western blot法检测海马组织中主要促进因子超家族成员2a(Mfsd2a)、紧密连接相关蛋白(Occludin)、血管生成素-1(Ang-1)、血管生成素-2(Ang-2)、内皮细胞TEK酪氨酸激酶(Tie2)蛋白表达情况。结果与假手术组比较,模型组大鼠第1~5天的逃避潜伏期均明显延长(P均<0.05),第6天和第7天大鼠穿过原平台次数明显减少、目标象限停留时间明显缩短(P均<0.05);脑组织中伊文思蓝含量明显增加(P<0.05),血脑屏障超微结构明显破坏,海马微血管密度明显增高(P<0.05);海马组织中Mfsd2a、Occludin蛋白相对表达量均明显降低(P均<0.05),Ang-1、Ang-2、Tie2蛋白相对表达量均明显增高(P均<0.05)。与模型组比较,尼莫地平组和通窍益智颗粒各组大鼠第1~5天的逃避潜伏期均明显缩短(P均<0.05),第6天和第7天大鼠穿过原平台次数明显增加、目标象限停留时间明显延长(P均<0.05);脑组织中伊文思蓝含量均明显降低(P均<0.05),血脑屏障超微结构破坏有不同程度的改善;尼莫地平组和通窍益智颗粒中、高剂量组大鼠海马微血管密度和海马组织中Mfsd2a、Occludin、Ang-1、Ang-2、Tie2蛋白相对表达量均明
Objective It is to investigate the effect of Tongqiao Yizhi granule on blood-brain barrier and angiogenesis in vascular dementia rats and its possible mechanism.Methods One hundred and thirty healthy clean grade male SD rats were selected,20 of them were randomly selected as the sham-operated group,and the remaining rats were established as vascular dementia models by permanent ligation of bilateral common carotid arteries.The successful modeled rats were randomly divided into model group,nimodipine group and high,medium and low dose groups of Tongqiao Yizhi granules,with 20 rats in each group.The nimodipine group was given nimodipine 6.25 mg/kg by gavage,the high,medium and low dose groups of Tongqiao Yizhi granules were given Tongqiao Yizhi granules 23.54 g/kg,11.77 g/kg and 5.88 g/kg by gavage,respectively,and the sham-operated and model groups were given equal amounts of normal saline by gavage.After 28 d of gavage,Morris water maze was used to assess the learning and memory ability of the rats in each group,Evans blue was used to assess the blood-brain barrier permeability,transmission electron microscopy was used to observe the ultrastructure of the blood-brain barrier,immunofluorescence labeling of CD31 protein in the CA1 region of the hippocampus was used to observe the microvessel density,and Western blot was used to detect the expression of major facilitator superfamily member 2a(Mfsd2a),tight junction related protein(Occludin),angiopoietin-1(Ang-1),angiopoietin-2(Ang-2),and endothelial TEK tyrosine kinase(Tie2)protein in hippocampal tissue.Results Compared with the sham-operated group,the evasion latency of rats in the model group was significantly prolonged on days 1-5(all P<0.05),and the number of crossing the original platform was significantly reduced,the dwell time in the target quadrant was significantly shortened on days 6 and 7(all P<0.05);the content of Evans blue in brain tissue was significantly increased(P<0.05),the ultrastructure of the blood-brain barrier was significantly disrupted,and t
作者
任俊豪
徐萍
李双阳
唐红梅
王饶琼
白雪
REN Junhao;XU Ping;LI Shuangyang;TANG Hongmei;WANG Raoqiong;BAI Xue(The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000,Sichuan,China;The Third People’s Hospital of Zigong City,Zigong 643000,Sichuan,China)
出处
《现代中西医结合杂志》
CAS
2022年第23期3238-3245,3334,共9页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
四川省中医药管理局科研专项(川中医药函【2020】196号)
泸州市科技局应用基础项目(2020-JYJ-57)
西南医科大学附属中医医院联合项目(2018XYLH-006)
西南医科大学附属中医医院科研项目(西南医大中医院【2019】119号)。
关键词
血管性痴呆
玄府
通窍益智颗粒
血脑屏障
血管新生
vascular dementia
Xuan Fu
Tongqiao Yizhi granule
blood-brain barrier
angiogenesis
