杜鹃花总黄酮通过抑制TNF-α/caspase-8/caspase-3信号通路减轻大鼠脑缺血再灌注损伤的作用  

The total flavonoids of rhododendron alleviate rat brain ischemia-reperfusion injury by inhibiting the TNF-α/caspase-8/caspase-3 signaling pathway

作　　者：余孝海[1] 金宇 孙敏琼 郭欠影 从辉 Yu Xiaohai;Jin Yu;Sun Minqiong;Guo Qianying;Cong Hui(School of Basic Medicine,Anhui Medical University,Hefei 230032;Dept of Pathology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Neurology,East Campus,The First Affiliated Hospital of Anhui Medical University,Hefei 231600)

机构地区：[1]安徽医科大学基础医学院,合肥230032 [2]安徽医科大学第一附属医院病理科,合肥230022 [3]安徽医科大学第一附属医院东城院区神经内科,合肥231600

出　　处：《安徽医科大学学报》2024年第6期1047-1052,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高校自然科学研究项目(编号:2023AH053310);安徽省研究生教育质量工程项目(编号:2022xscx053);安徽医科大学国家级大学生创新创业训练计划项目(编号:202310366037)。

摘　　要：目的探索杜鹃花总黄酮(TFR)通过抑制TNF-α/caspase-8/caspase-3信号通路保护脑缺血再灌注(I/R)损伤的机制。方法采用大脑中动脉闭塞术(MCAO)建立大鼠I/R模型,将造模成功大鼠随机分为假手术(Sham)、脑缺血再灌注(MCAO)和脑缺血再灌注术后TFR 200 mg/kg干预(TFR 200 mg/kg)组,制备MCAO大鼠模型,在脑缺血再灌注损伤手术后,TFR 200 mg/kg组连续14 d给予TFR(200 mg/kg)药物溶液。术后14 d,依据大鼠神经功能评分,脑血流图观察脑血流情况,处死大鼠,苏木精-伊红(HE)染色观察脑组织细胞形态学变化,试剂盒检测大鼠血清中乳酸脱氢酶(LDH或LD)和神经特异性烯醇化酶(NSE)两种酶活性;ELISA试剂盒检测白细胞介素-1(IL-1)和IL-6水平,Western blot和免疫组化同时检测大鼠脑组织中裂解型胱天蛋白酶3(cleaved caspase-3)、caspase-8、肿瘤坏死因子(TNF)-α蛋白的表达水平。结果脑缺血再灌注处理后,MCAO导致了大鼠神经功能异常,神经功能评分指数显著升高,脑组织病理形态学和脑血流量变化明显,脑组织中cleaved caspase-3、caspase-8、TNF-α蛋白的表达水平显著增加,血清中LDH、NSE、IL-1、IL-6水平明显升高;TFR 200 mg/kg组大鼠神经功能评分明显降低,脑组织病理损伤显著改善,脑组织中cleaved caspase-3、caspase-8、TNF-α蛋白的表达以及血清中LDH、NSE、IL-1、IL-6水平降低。结论TFR可能通过抑制TNF-α/caspase-8/caspase-3信号通路减轻脑缺血缺氧性损伤。Objective To explore the mechanism by which total flavones of rhododendron(TFR)protect against cerebral ischemia-reperfusion(I/R)injury by inhibiting the TNF-α/caspase-8/caspase-3 signaling pathway.Methods The middle cerebral artery occlusion(MCAO)method was used to establish the rat I/R model.Rats were randomly divided into Sham surgery,MCAO,and post-I/R intervention with TFR 200 mg/kg(TFR 200 mg/kg)groups.After establishing the MCAO rat model,rats in the TFR 200 mg/kg group were administered TFR(200 mg/kg)solution for 14 consecutive days following I/R injury surgery.Hematoxylin-Eosin(HE)staining was used to observe neurological function scoring,cerebral blood flow assessment,histological examination of brain tissue,assay kits were used to detect lactate dehydrogenase(LDH)and neuron-specific enolase(NSE)activities in rat serum.ELISA assay kits was used to measure interleukin-1(IL-1)and interleukin-6(IL-6)levels,and Western blot and immunohistochemistry were conducted to detect the expression levels of cleaved caspase-3,caspase-8,and TNF-αproteins in rat brain tissue 14 days post-surgery.Results After cerebral ischemia-reperfusion treatment,MCAO resulted in abnormal neurological function in rats,significantly increased neurological function scoring index,obvious changes in cerebral tissue histomorphology and cerebral blood flow,significant upregulation of cleaved caspase-3,caspase-8,and TNF-αprotein expression levels in brain tissue,and significant elevation of LDH,NSE,IL-1,and IL-6 levels in serum.Rats in the TFR 200 mg/kg group showed significantly reduced neurological function scoring,significant improvement in cerebral tissue pathological damage,decreased expression levels of cleaved caspase-3,caspase-8,and TNF-αproteins in brain tissue,as well as decreased levels of LDH,NSE,IL-1,and IL-6 in serum.Conclusion TFR may alleviate cerebral ischemic hypoxic injury by inhibiting the TNF-α/caspase-8/caspase-3 signaling pathway.

关 键 词：杜鹃花总黄酮 脑缺血再灌注 脑缺血缺氧性损伤 凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]