丹蒌片对Aβ1-40致脑微血管内皮细胞损伤保护作用的机制研究  

Protective Mechanism of Dan Lou Tablets on Aβ1-40-induced Brain Microvascular Endothelial Cell Damage

作　　者：高爱社[1] 杨洒 张振强[2] 宋军营 张鹏飞[1] 史占江 GAO Aishe;YANG Sa;ZHANG Zhenqiang;SONG Junying;ZHANG Pengfei;SHI Zhanjiang(School of Basic Medical Sciences of Henan University of Chinese Medicine,Zhengzhou 450018 Henan,China;Academy of Chinese Medicine Sciences of Henan University of Chinese Medicine,Zhengzhou 450018 Henan,China)

机构地区：[1]河南中医药大学基础医学院,河南郑州450018 [2]河南中医药大学中医药科学院,河南郑州450018

出　　处：《中药新药与临床药理》2021年第2期234-239,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然基金联合项目(U1504829);中原科技领军人才项目(204200510022);河南省高等学校重点科研项目(20A310009);河南中医药大学博士基金项目(BSJJ2018-01);河南省科技攻关项目(202102310508)。

摘　　要：目的研究丹蒌片对β淀粉样蛋白1-40(Aβ1-40)诱导脑微血管内皮细胞(BMEC)损伤的保护作用。方法(1)将大耳白兔BMEC培养在含10%胎牛血清和1%青霉素/链霉素的完全培养基中,用Aβ1-40建立BMEC损伤模型。实验分正常组(C)、模型组(M)和晚期糖基化终末产物受体(RAGE)中和抗体组(RAGE-Ab),24 h后使用Western Blot法检测各组血管细胞黏附分子-1(VCAM-1)、巨噬细胞炎性蛋白-1β(MIP-1β)、趋化因子受体-5(CCR5)的表达。(2)用丹蒌片制备含药血清,实验分为正常组(10%空白血清培养)、模型组、低浓度用药组(10%丹蒌片低浓度含药血清+Aβ1-40)、高浓度用药组(10%丹蒌片高浓度含药血清+Aβ1-40),24 h后用Western Blot法测定各组细胞上述因子的表达水平。结果(1)与正常组相比,模型组RAGE、VCAM-1、MIP-1β、CCR5表达显著增加(P<0.01),RAGE-Ab组各因子表达水平显著降低(P<0.01);(2)与空白血清组相比,模型组RAGE、VCAM-1、MIP-1β、CCR5表达增加(P<0.001),与模型组相比,丹蒌片含药血清组各因子的表达显著降低(P<0.01)。结论丹蒌片含药血清具有保护Aβ1-40诱导的BMEC损伤的作用,其机制可能与抑制Aβ和RAGE诱发的BMEC趋化反应有关。Objective To explore the protective effect of Dan Lou tablets on Aβ1-40)-induced brain microvascular endothelial cells(BMEC)injury.Methods Rabbit BMEC were cultured in a complete medium containing 10%fetal bovine serum and 1%penicillin/streptomycin,and divided into normal control group(C),model group(M Aβ1-40model)and advanced glycosyl End product receptor(RAGE)neutralizing antibody group(RAGE-Ab),24 hours later,Western Blot was used to detect vascular cell adhesion molecule-1(VCAM-1),macrophage inflammatory protein-1(3(MIP-1β),and the expression of chemokine receptor-5(CCR5).BMEC was cultured with Dan Lou tablets medicated serum and divided into normal group(10%blank serum culture),model group(Aβ1-40 to establish BMEC injury model),and low concentration drug group(10%Dan Lou tablets low concentration medicated serum+Aβ1-40),high-concentration drug group(10%Dan Lou tablets high-concentration medicated serum+Aβ1-40,24 hours later,the expression levels of the above factors in the cells of each group were determined by Western Blot.Results Compared with the normal group,the expression of RAGE,VCAM-1,MIP-1βand CCR5 in the model group increased significantly(P<0.01),and the expression levels of various factors in the RAGE-Ab group decreased significantly(P<0.01).Compared with the blank serum group,the expression of RAGE,VCAM-1,MIP-1βand CCR5 in the model group increased significantly(P<0.01).Compared with the model group,the expression of RAGE,VCAM-1,MIP-1βand CCR5 increased(P<0.001).Compared with the model group,the expression of each factor in the serum group containing Dan Lou tablets was significantly reduced(P<0.01).Conclusion The serum containing Dan Lou tablets can protect BMEC from damage induced by Aβ1-40.The mechanism may be related to the inhibition of Aβ/RAGE-induced BMEC chemotaxis.

关 键 词：丹蒌片 脑微血管内皮细胞 Β淀粉样蛋白1-40 阿尔茨海默病 兔 

分 类 号：K285.5[历史地理—中国史]