基于NF-κB通路研究平肝活血合剂对脑缺血再灌注损伤的作用机制  

Mechanism of Pinggan Huoxue mixture on cerebral ischemia reperfusion injury based on the NF-κB pathway

作　　者：刘宗梅 王晓玲[2] 宋发萍 姚硕 唐勇[2] LIU Zong-mei;WANG Xiao-ling;SONG Fa-ping;YAO Shuo;TANG Yong(The First Clinical Medical College,Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046;Ankang Hospital of Traditional Chinese Medicine,Ankang Shaanxi 725000)

机构地区：[1]陕西中医药大学第一临床医学院,陕西咸阳712046 [2]安康市中医医院,陕西安康725000

出　　处：《中南药学》2024年第4期902-907,共6页Central South Pharmacy

基　　金：陕西省中医药管理局课题(No.2021-GJ-JC011);陕西省2021年度中医药传承创新暨“秦药”开发重点科学研究项目(No.2021-01-GJ-019);第七批全国老中医药专家学术经验继承工作项目。

摘　　要：目的 探讨平肝活血合剂对脑缺血再灌注损伤(CIRI)的作用机制。方法 采用线栓法制备CIRI模型。SD大鼠随机分为假手术组,模型组,平肝活血合剂低、中、高剂量组(2.08、4.16、8.32 mL·kg^(-1)),阳性对照组(丁苯酞软胶囊6.25 mg·kg^(-1)),每组10只。灌胃治疗7d,每日1次。造模后第1、3、7日观察大鼠神经功能评分。给药7日后观察大鼠脑缺血组织病理改变,测定脑梗死体积百分比,检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)的水平,脑缺血组织中的TNF-α、IL-1β、IL-8 mRNA表达,脑缺血组织中磷酸化的NF-κB抑制蛋白α(p-IκBα)、核转录因子-κBp65(NF-κBp65)、磷酸化的核转录因子-κBp65(p-NF-κBp65)蛋白的表达。结果 与假手术组比较,模型组大鼠给药7日后Longa评分显著升高,右脑出现鲜明的苍白色梗死灶(P <0.01),TNF-α、IL-1β、IL-8及TNF-α、IL-1β、IL-8 mRNA表达均显著上升,p-IκBα、NF-κBp65、p-NF-κBp65蛋白表达均明显上调(P <0.05或P <0.01)。HE染色显示脑缺血组织区网状结构疏松,细胞错乱、结构缺损甚至萎缩变形、数量变少,核固缩及碎裂。与模型组比较,给药7日后平肝活血合剂各剂量组Longa评分显著降低,其余检测指标亦明显下降(P <0.05或P <0.01)。结论 平肝活血合剂可显著改善CIRI的炎症因子水平,减轻脑损伤,其机制可能与调控NF-κB信号通路有关。Objective To determine the mechanism of Pinggan Huoxue mixture on cerebral ischemia-reperfusion injury(CIRI).Methods CIRI model was induced by suture-occlusion method.SD rats were randomly divided into a sham operation group,a model group,Pinggan Huoxue mixture low-dose,-medium-dose,and high-dose groups(2.08,4.16,and 8.32 mL·kg^(-1)),and a positive control group-(butylphthalein soft capsules at 6.25 mg·kg^(-1)),with 10 rats in each group.Each group was given water or drug intragastrically,once a day,for consecutive 7 days.The neurological function scores was observed on the 1st,3rd,and 7th day after the modeling.After 7 days of administration,the pathological changes in rats cerebral ischemic tissue were observed,and the percentage of cerebral infarction volume was measured.The serum tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-8 levels and TNF-α,IL-1β,IL-8 mRNA expression,and NF-κB inhibitory proteinα(p-IκBα),nuclear transcription factor-κBp65(NF-κBp65),phosphorylated nuclear transcription factors-κBp65(p-NF-κBp65)protein expression in the ischemic brain tissue were also detected.Results Compared with the sham operation group,the Longa score of the model group significantly increased after 7 days of administration,and a distinct pale infarct lesion appeared in the right brain(P<0.01).TNF-α,IL-1β,IL-8 and TNF-α,IL-1β,IL-8 mRNA expressions were significantly increased(P<0.01),while the protein expressions of p-IκBα,NF-κBp65 and p-NF-κBp65 were significantly upregulated(P<0.05 or P<0.01).HE staining showed a loose network structure in the ischemic tissue area of the brain,with disordered cells,structural defects,and even atrophy and deformation,reduced quantity,nuclear pyknosis and fragmentation.Compared with the model group,after 7 days of administration,the Longa scores of rats in each dose groups on the 7th day of intervention were significantly decreased,and other detection indicators were also decreased(P<0.05 or P<0.01).Conclusion Pinggan Huoxue mixture can greatly improve t

关 键 词：平肝活血合剂 脑缺血再灌注损伤 炎症抑制 NF-ΚB信号通路 

分 类 号：R285[医药卫生—中药学]