通过TLR4/NF-κB通路探讨白藜芦醇对急性脑出血大鼠脑损伤的作用机制  

Mechanism of Resveratrol on Brain Injury in Rats with Acute Cerebral Hemorrhage through TLR4/NF-κB Pathway

作　　者：杨龙[1] 董秀娟[1] YANG Long;DONG Xiujuan(Cangzhou Central Hospital,Cangzhou 061000,Hebei,China)

机构地区：[1]沧州市中心医院,河北沧州061000

出　　处：《中西医结合心脑血管病杂志》2023年第12期2181-2187,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河北省医学科学研究课题项目(No.20201544)。

摘　　要：目的:观察白藜芦醇(RES)对急性脑出血大鼠脑组织损伤的保护作用及其机制。方法:将240只健康雄性Wistar大鼠随机分为假手术组、模型组、RES低剂量组(30 mg/kg)、RES中剂量组(60 mg/kg)、RES高剂量组(90 mg/kg)和尼莫地平组(NMP组,10 mg/kg),每组40只。采用自体血注射法制备急性脑出血大鼠模型,造模完成后,各组分别腹腔注射给药(假手术组和模型组给予0.9%氯化钠溶液),每日1次,共7 d。采用Longa评分标准进行神经功能缺损评分,失重法检测脑组织含水量,测定脑血肿体积;采用苏木精-伊红(HE)染色法、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)分别观察脑组织病理学改变和神经细胞凋亡;酶联免疫吸附法(ELISA)检测脑组织白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量;分光光度法检测脑组织丙二醛(MDA)含量和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性;免疫组织化学(IHC)法检测脑组织离子钙结合适配分子1(Iba1)表达;蛋白免疫印迹法(Western Blot)检测脑组织Toll样受体4(TLR4)、核因子-κB p65(NF-κB p65)表达。结果:与假手术组比较,模型组大鼠神经功能缺失评分、脑含水量、血肿体积升高(P<0.01);血肿周围脑组织可见大量红细胞、脑组织水肿、神经细胞减少、细胞核固缩、炎性细胞浸润等病理改变;凋亡细胞数量增多,凋亡指数(AI)升高(P<0.01);脑组织IL-1β、IL-6、TNF-α、MDA含量升高,SOD、CAT活性降低(P<0.01);小胶质细胞形态呈肥大阿米巴虫样,Iba1表达量、脑组织TLR4、NF-κB p65表达量升高(P<0.01)。与模型组比较,RES中剂量组、RES高剂量组和NMP组神经功能缺失评分、脑含水量、脑血肿体积降低(P<0.05或P<0.01);脑组织病理学改变和神经细胞凋亡状况均改善,AI降低(P<0.01);脑组织IL-1β、IL-6、TNF-α、MDA含量降低,SOD、CAT活性升高(P<0.05或P<0.01);小胶质细胞形态呈瘦长分�Objective:To observe the protective effect of resveratrol(RES)on brain tissue injury in rats with acute cerebral hemorrhage.Methods:Two hundred and forty healthy Wistar rats were randomly divided into sham operation group,model group,RES low-dose group(30 mg/kg),RES medium-dose group(60 mg/kg),RES high-dose group(90 mg/kg)and Nimodipine group(NMP group,10 mg/kg),with 40 rats in each group.The model of rats with acute cerebral hemorrhage were established by autologous blood injection.After modeling,each group was treated with intraperitoneal injection(sham operation group and model group were treated with 0.9%sodium chloride solution),once a day for 7 days.The neurological deficit was determined according to the Longa score,the water content of brain tissue was detected by weightlessness method,and the volume of cerebral hematoma was determined.The pathological changes of brain tissue was detected by hematoxylin-eosin(HE)staining method,the nerve cell apoptosis was detected by termina-l deoxynucleotidyl transferase mediated nick end labeling(TUNEL).The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in brain tissue were detected by enzyme-linked immunosorbent assay(ELISA).The content of malondialdehyde(MDA),the activities of superoxide dismutase(SOD)and catalase(CAT)were detected by spectrophotometry.The expression of ionized calcium binding adaptor molecule 1(Iba1)in brain tissue was detected by immunohistochemistry(IHC).The expressions of Tol-l like receptor 4(TLR4)and nuclear facto-rκB p65(NF-κB p65)were detected by Western Blot.Results:Compared with sham operation group,neurological deficit score,brain water content,and hematoma volume were increased in the model group(P<0.01).Pathological changes such red blood cells,edema of brain tissue,decrease of nerve cells,nucleolysis and inflammatory cell infiltration were seen in the brain tissue around hematoma.The number of apoptotic cells,and the apoptotic index(AI)were significantly increased(P<0.01).The contents of IL-1

关 键 词：急性脑出血 白藜芦醇 炎症 氧化应激 Toll样受体4/核因子-κB通路 大鼠 实验研究 

分 类 号：R285.5[医药卫生—中药学]