主 办:北 京 中 医 药 大 学
ISSN 2095-6606 CN 10-1157/R

现代中医临床 ›› 2021, Vol. 28 ›› Issue (3): 28-33.doi: 10.3969/j.issn.2095-6606.2021.03.007

• 中医临床基础研究 • 上一篇    下一篇

清开灵注射液对Aβ42刺激下小鼠体外血脑屏障转运Aβ的影响及机制研究*

窦晋芳1, 常佩芬2, 胡超群2, 高瑜倩2, 张洪嘉2, 彭嘉冉2, 王志淼2, 黑穆荣2, 高永红1, 郭宁2, 朱海燕1,2,3#   

  1. 1 北京中医药大学东直门医院 中医内科学教育部和北京市重点实验室 北京 100700;
    2 北京中医药大学东直门医院心内科;
    3 北京中医药大学心血管病研究所
  • 收稿日期:2020-10-10 出版日期:2021-05-30 发布日期:2021-06-08
  • 通讯作者: #朱海燕,女,博士,主任医师,硕士生导师, E-mail:zhuhaiyangy@163.com
  • 作者简介:窦晋芳,女,在读硕士生
  • 基金资助:
    *国家自然科学基金项目(No.81202686, No.81874445)

Effects of Qingkailing Injection on blood-brain barrier in vitro and transport of β-amyloid in mice stimulated by Aβ42 and its mechanism*

Dou Jinfang1, Chang Peifen2, Hu Chaoqun2, Gao Yuqian2, Zhang Hongjia2, Peng Jiaran2, Wang Zhimiao2, Hei Murong2, Gao Yonghong1, Guo Ning2, Zhu Haiyan1,2,3#   

  1. 1 Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700;
    2 Department of Cardiology, Dongzhimen Hospital, Beijing University of Chinese Medicine;
    3 Institute of Cardiovascular Disease, Beijing University of Chinese Medicine
  • Received:2020-10-10 Online:2021-05-30 Published:2021-06-08

摘要: 目的 观察具有神经毒性的寡聚肽Aβ42对体外血脑屏障通透性及转运Aβ的影响以及清开灵的干预作用。方法 通过体外培养 Balb/c小鼠脑微血管内皮细胞形成血脑屏障,随机分为正常对照组(加入 DMEM)、模型组(加入 Aβ42)和清开灵组(加入 Aβ42+清开灵),测定各组TEER值、HPR 2小时通透率,用ELISA法检测各孔Aβ40浓度值,用Western blot法检测RAGE、LRP-1、Claudin-5、Occludin、PKC和ERK的表达水平。结果 与正常组比较,模型组TEER值、claudin-5的表达显著降低(P<0.05,0.01),HRP、Aβ40的透过量、RAGE、PKC蛋白的表达量明显增加(P<0.01);与模型组比较,清开灵能够增加 TEER 值(P<0.05)、claudin-5的表达(P<0.01),抑制HPR和Aβ40的透过量(P<0.05,0.01),明显减少RAGE、PKC蛋白表达(P<0.01)。结论 清开灵注射液对血脑屏障的结构和功能均有保护作用,通过明显减少RAGE表达,增加Claudine-5表达,分别从细胞内转运途径和细胞旁转运途径抑制Aβ的透过,阻断AD发病过程中的恶性循环,这主要与抑制PKC信号通路有关。

关键词: 清开灵注射液, 血脑屏障, β-淀粉样蛋白, 阿尔茨海默病

Abstract: Objective To observe the effects of neurotoxic Aβ42 on the permeability and Aβ transportation of blood-brain barrier(BBB) in vitro and the role of Qingkailing Injection. Methods The BBB in vitro was established by culturing cerebral microvasculature endothelial cells of Balb/c mice and then they were divided into the normal group, model group and Qingkailing group. The transendothelial electrical resistance (TEER) and the permeability to HRP were used as indicators of changes in BBB permeability. The concentration of Aβ40 and protein expression levels of RAGE, LRP-1, Claudin-5, Occludin, PKC and ERK were measured by enzyme-linked immuno sorbent assays and western blotting respectively. Results As compared with control group, the TEER and the expressions of claudin-5 in BBB decreased significantly in the model group(P<0.05,0.01), while the permeability to HRP, the concentration of Aβ40, the expression of RAGE and PKC increased (P<0.01). Compared with model group, Qingkailing effectively decreased the permeability of the BBB(P<0.05,0.01), promoted the expression of claudin-5 (P<0.01) and down-regulated the expressions of RAGE and PKC (P<0.01). Conclusions Qingkailing Injection has a protective effect on the structure and function of BBB. It suppresses the permeability to Aβ40 through intracellular and extracellular transport pathways respectively, by down-regulating the expression of RAGE and up-regulating the expression of claudin-5, breaking the vicious cycle of AD. This beneficial effect may be linked to the inhibition of the PKC signal pathways.

Key words: Qingkailing Injection, blood-brain barrier, β-amyloid, alzheimer disease

中图分类号: 

  • R259