甲基莲心碱通过调节CCL2-CCR2信号通路对缺血性脑卒中大鼠血脑屏障的影响  

Effect of neferine on blood-brain barrier in rats with ischemic stroke by regulating CCL2-CCR2 signal pathway

作　　者：翟春晓 田平[2] 李豪 孟贝[1] ZHAI Chunxiao;TIAN Ping;LI Hao;MENG Bei(School of Nursing,Xinyang Vocational and Technical College,Xinyang 464000,China;Department of Scientific Research,Xinyang Polytechnic,Xinyang 464000,China;Clinical Medicine,Xinyang Maternal and Child Health Hospital,Xinyang 464000,China)

机构地区：[1]信阳职业技术学院护理学院,信阳464000 [2]信阳职业技术学院科研处,信阳464000 [3]信阳市妇幼保健院临床医学,信阳464000

出　　处：《中国免疫学杂志》2024年第6期1166-1171,共6页Chinese Journal of Immunology

基　　金：河南省科技攻关项目(222102310520)。

摘　　要：目的:探究甲基莲心碱(NEF)调节CC趋化因子配体2(CCL2)-趋化因子受体2(CCR2)信号通路对缺血性脑卒中(IS)大鼠血脑屏障的影响。方法:将雄性SD大鼠随机分为模型组(Model组)、低剂量NEF组(NEF-L组,25 mg/kg NEF)、高剂量NEF组(NEF-H组,50mg/kgNEF)和高剂量NEF+CCR2拮抗剂组(NEF-H+RS102895组,50mg/kgNEF+10mg/kg RS102895)和假手术组(Sham组),每组15只。比较各组大鼠神经功能评分。伊文思蓝(EB)渗透法评价各组大鼠血脑屏障通透性。采用干湿重法测定各组大鼠脑组织含水量。实时荧光定量PCR(RT-qPCR)测定大鼠脑组织CCL2、CCR2 mRNA表达。Western blot检测大鼠脑组织CCL2、CCR2、ZO-1、Claudin-5蛋白表达。结果:与Sham组相比,Model组大鼠神经功能评分、右脑组织EB含量、含水量、脑组织CCL2、CCR2 mRNA和蛋白表达显著增加(P<0.05),ZO-1、Claudin-5水平显著降低(P<0.05)。相较于Model组,NEF-H组神经功能评分、脑含水量、EB含量、CCL2、CCR2 mRNA和蛋白水平显著降低(P<0.05),ZO-1、Claudin-5表达水平显著升高(P<0.05)。RS102895增强了NEF对IS大鼠血脑屏障的保护作用。结论:NEF可能通过抑制CCL2-CCR2信号通路对IS大鼠血脑屏障起保护作用。Objective:To investigate the impact of neferine(NEF)on the blood-brain barrier in ischemic stroke(IS)rats by regulating the CC chemokine ligand 2(CCL2)-CC chemokine receptor 2(CCR2)signal pathway.Methods:SD rats were randomly divided into model group,low-dose NEF group(NEF-L group,25 mg/kg NEF),high-dose NEF group(NEF-H group,50 mg/kg NEF),and high-dose NEF+CCR2 antagonist group(NEF-H+RS102895 group,50 mg/kg NEF+10 mg/kg RS102895),sham opera-tion group(Sham group),with 15 rats in each group.The neurological function scores of rats in each group were compared.The perme-ability of blood-brain barrier of rats in each group was evaluated by Evans blue(EB)permeation method.The water content of brain tis-sue of rats in each group was determined by dry and wet weight method.The expression levels of CCL2 and CCR2 mRNA in rat brain tissue were measured by real-time fluorescence quantitative PCR(RT-qPCR).The expression levels of CCL2,CCR2,ZO-1 and Clau-din-5 proteins in rat brain tissue were detected by Western blot.Results:Compared with Sham group,the neural function score,EB content,water content,the expression levels of CCL2,CCR2 mRNA and protein in the right brain tissue of the model group were obvi-ously higher(P<0.05),the expression levels of ZO-1 and Claudin-5 were lower(P<0.05).Compared with the model group,the neuro-logical function score,water content,EB content,the expression levels of CCL2,CCR2 mRNA and protein in the right brain tissue of rats in the NEF-H group were obviously lower(P<0.05),the expression levels of ZO-1 and Claudin-5 were obviously higher(P<0.05).RS102895 enhanced the protective effect of NEF on the blood-brain barrier in IS rats.Conclusion:NEF may protect the blood-brain barrier of IS rats by inhibiting CCL2-CCR2 signal pathway.

关 键 词：甲基莲心碱 CC趋化因子配体2-趋化因子受体2信号通路 缺血性脑卒中 血脑屏障 

分 类 号：R285.5[医药卫生—中药学] R743.3[医药卫生—中医学]