没食子酸靶向β-arrestin2抑制星形胶质细胞炎症的机制研究  

Mechanism Study of Gallic Acid Targetingβ-arrestin2 to Inhibit Astrocyte Inflammation

作　　者：夏行宇 王东硕 林彬燕 朱琴 XIA Xingyu;WANG Dongshuo;LIN Binyan;ZHU Qin(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;TCM Pharmacy,Langfang Traditional Chinese Medicine Hospital,Langfang 065000,China;Experiment Center for Science and Technology,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]廊坊市中医医院中药房,河北廊坊065000 [3]南京中医药大学科技实验中心,江苏南京210023

出　　处：《南京中医药大学学报》2025年第3期341-351,共11页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：江苏省高等学校基础科学(自然科学)面上项目(21KJB360015)。

摘　　要：目的探究没食子酸靶向β-arrestin2抑制星形胶质细胞炎症的机制。方法通过PharmMapper寻找没食子酸的潜在靶点并通过热迁移和分子对接实验验证。采用Western blot检测β-arrestin2、NF-κB信号通路和NLRP3炎症小体相关蛋白表达;qPCR检测IL-1β、IL-6和TNF-α等促炎细胞因子的mRNA水平。构建MPTP诱导的亚急性PD小鼠模型,通过开场实验、转棒实验和爬杆实验评估小鼠的行为学能力;通过免疫组化检测小鼠SNc区TH^(+)和GFAP^(+)细胞数目;Western blot检测小鼠中脑组织匀浆蛋白中NF-κB信号通路和NLRP3炎症小体相关蛋白的表达。结果分子对接实验和热迁移实验表明没食子酸可以直接与β-arrestin2结合。没食子酸可以降低星形胶质细胞中p-IKK和p-P65蛋白表达(P<0.001,P<0.0001),降低IL-1β、IL-6和TNF-α的mRNA水平(P<0.05,P<0.01),降低caspase-1和IL-1β蛋白表达(P<0.0001),但对β-arrestin2蛋白表达没有影响(P>0.05)。没食子酸可以改善PD模型小鼠行为能力,增加PD模型小鼠TH^(+)神经元数和GFAP^(+)细胞数(P<0.05,P<0.001),减少PD模型小鼠中脑caspase-1、IL-1β、NLRP3和p-IKK蛋白表达(P<0.05)。结论没食子酸通过结合β-arrestin2抑制NF-κB信号通路的激活和NLRP3炎症小体的活化以减轻星形胶质细胞炎症,并且对MPTP诱导的PD模型小鼠具有神经保护作用。OBJECTIVE To explore the mechanism of gallic acid targetingβ-arrestin2 to inhibit astrocyte inflammation.METHODS Potential targets of gallic acid were found by PharmMapper and verified by thermal shift and molecular docking experiments.Western blot was used to detect the expression ofβ-arrestin2,NF-κB signaling pathway and NLRP3 inflammasome-related proteins;qPCR was used to detect the mRNA levels of proinflammatory cytokines such as IL-1β,IL-6 and TNF-α.A subacute PD mouse model induced by MPTP was established,and the behavioral ability of mice was evaluated by open field test,rotating rod test and climbing pole test;the number of TH^(+)and GFAP^(+)cells in the SNc area of mice was detected by immunohistochemistry;Western blot was used to detect the expression of NF-κB signaling pathway and NLRP3 inflammasome-related proteins in the homogenate protein of mouse midbrain tissue.RESULTS Molecular docking experiments and thermal shift experiments showed that gallic acid could directly bind toβ-arrestin2.Gallic acid could reduce the expression of p-IKK and p-P65 proteins in astrocytes(P<0.001,P<0.0001),reduce the mRNA levels of IL-1β,IL-6 and TNF-α(P<0.05,P<0.01),and reduce the expression of caspase-1 and IL-1βproteins(P<0.0001),but had no effect on the expression ofβ-arrestin2 protein(P>0.05).Gallic acid could improve the behavioral ability of PD model mice,increase the number of TH^(+)neurons and GFAP^(+)cells in PD model mice(P<0.05,P<0.001),and reduce the expression of caspase-1,IL-1β,NLRP3 and p-IKK proteins in the brain of PD model mice(P<0.05).CONCLUSION Gallic acid inhibits the activation of NF-κB signaling pathway and NLRP3 inflammasome by binding toβ-arrestin2 to reduce astrocyte inflammation,and has a neuroprotective effect on MPTP-induced PD model mice.

关 键 词：β-arrestin2 没食子酸 星形胶质细胞 神经炎症 NF-ΚB信号通路 NLRP3炎症小体 

分 类 号：R285.5[医药卫生—中药学]