基于网络药理学的白芍总苷改善大鼠脑出血后神经炎症的作用及机制研究  

Effect and Mechanism of Total Glucosides of Paeony in Ameliorating Neuroinflammation after Cerebral Hemorrhage in Rats Based on Network Pharmacology

作　　者：朱军 洪明奕 石程吉 刘玉洲 蔡昱 邢晋山 徐厚平 顾应江 ZHU Jun;HONG Mingyi;SHI Chengji;LIU Yuzhou;CAI Yu;XING Jinshan;XU Houping;GU Yingjiang(Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000;The Affiliated Stomatological Hospital of Southwest Medical University,Luzhou 646000)

机构地区：[1]西南医科大学附属中医医院,泸州646000 [2]西南医科大学附属口腔医院,泸州646000

出　　处：《中药药理与临床》2023年第12期70-78,共9页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川省科技厅项目(编号:2019YFS0543)。

摘　　要：目的:本研究通过网络药理学和动物实验探究白芍总苷对脑出血(Intracerebral hemorrhage,ICH)后神经炎症的脑保护作用及可能的作用机制。方法:通过网络药理学分析获得白芍总苷治疗脑出血的可能靶点和信号通路。动物实验将60只大鼠随机分为正常对照组、模型对照组、白芍总苷0.05、0.1、0.2 g/kg组。采用胶原酶诱导大鼠脑出血模型,大鼠进行改良神经功能缺损评分(Modified neurological severity score,mNSS);HE染色观察血肿周围脑组织结构病理形态学改变;免疫荧光(IF)检测小胶质细胞M1型标记物CD16^(+)、M2型标记物CD206^(+)的表达变化;Western blot法检测大鼠脑组织肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-10、P38、p-P38蛋白的表达。结果:网络药理学分析筛选出TNF、IL-6、MMP9、VEGFA、EGFR、MAPK1是白芍总苷治疗ICH的核心作用靶点,通过GO和KEGG通路富集分析,MAPK信号通路是其中较为重要的途径;分子对接显示芍药苷、羟基芍药苷、芍药内酯苷、苯甲酰芍药苷与IL-6及TNF-α等关键靶点有较强亲和能力。动物实验结果表明,与正常对照组相比,模型对照组大鼠改良神经功能缺损评分明显升高(P<0.05),HE染色显示脑组织病理损伤明显加重,CD16^(+)、CD206^(+)的比例明显增加(P<0.05),大鼠脑组织IL-6、TNF-α、IL-10、p-P38/P38蛋白表达明显上调(P<0.05);与模型对照组相比,给药后,大鼠改良神经功能缺损评分明显降低(P<0.05),HE染色显示脑组织病理状况得以改善,CD16^(+)的比例明显降低,CD206^(+)的比例明显增加(P<0.05),除白芍总苷0.05 g/kg组中磷酸化P38蛋白表达无变化外,其余各给药组大鼠脑组织IL-6、TNF-α、p-P38/P38蛋白表达明显下调,IL-10蛋白表达明显上调(P<0.05)。结论:白芍总苷能减轻胶原酶诱导的脑出血大鼠神经炎症反应,抑制M1型小胶质细胞过度激活,同时促进其向M2型小胶质细胞转化,其机制可能与抑制P38MAPK�Objective:To investigate the neuroprotective effect of total glucosides of paeony(TGP)on post-intracerebral hemorrhage(ICH)neuroinflammation and explore potential mechanism based on network pharmacology and animal experiments.Methods:Network pharmacology was used to identify potential targets and signaling pathways of TGP in treating ICH.Sixty rats were randomly divided into a normal control group,a model control group,and TGP groups of different doses(0.05,0.1,and 0.2 g/kg).A collagenase-induced ICH model was established in rats,and the rats underwent modified neurological severity score(mNSS)evaluation.Hematoxylin-eosin(HE)staining was performed to observe pathological changes in brain tissues around the hematoma.Immunofluorescence(IF)was used to detect the expression of M1-type marker CD16^(+)and M2-type marker CD206^(+)in microglia.Western blot was used to measure the expression of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-10,P38,and phosphorylated P38(p-P38)proteins in rat brain tissues.Results:Network pharmacology identified TNF,IL-6,MMP9,VEGFA,EGFR,and MAPK1 as core targets of TGP in treating ICH.The MAPK signaling pathway was shown to be an important pathway through GO and KEGG pathway enrichment analysis.Molecular docking revealed strong binding affinity of paeoniflorin,hydroxypaeoniflorin,paeoniflorin lactone glycoside,and benzoylpaeoniflorin to key targets such as IL-6 and TNF-α.In animal experiments,compared with the normal control group,the model control group showed a significant increase in mNSS scores(P<0.05),exacerbated pathological damage in brain tissues based on HE staining,increased proportions of CD16^(+)microglia,increased proportions of CD206^(+)microglia(P<0.05),and upregulated protein expression of IL-6,TNF-α,IL-10,and p-P38/P38 in rat brain tissues(P<0.05).After TGP administration,the TGP groups showed decreased mNSS scores(P<0.05),improved brain tissue pathological conditions by HE staining,decreased CD16^(+)microglia proportions,and increased CD206^(+)microglia proport

关 键 词：白芍总苷 网络药理学 脑出血 神经炎症 小胶质细胞 

分 类 号：R285.5[医药卫生—中药学]