原花青素通过抑制Toll样受体4/核因子-κB信号通路对大鼠脑缺血再灌注损伤的保护作用  

Protective effect of procyanidins on cerebral ischemia-reperfusion injury in ratsby inhibiting TLR4/NF-κB signaling pathway

作　　者：任娟 覃宏伟 周冬亮 张敏杰 徐瑛 REN Juan;QING Hongwei;ZHOU Dongliang;ZHANG Minjie;XU Ying(Department of Neurology,Renhe Hospital of Baoshan District Shanghai,Shanghai 200431,China)

机构地区：[1]上海市宝山区仁和医院神经内科,上海200431

出　　处：《安徽医药》2021年第3期431-435,I0001,共6页Anhui Medical and Pharmaceutical Journal

基　　金：上海市宝山区仁和医院重点学科建设计划(YXK-2018-01A)。

摘　　要：目的探究原花青素通过抑制Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路对大鼠脑缺血再灌注(ischemia-reper⁃fusion,I/R)损伤的保护作用。方法选取60只雄性SD大鼠,采用随机数字表法分为对照组、模型组、原花青素低剂量组(80 mg/kg)和原花青素高剂量组(160 mg/kg),每组15只;除对照组外,其他各组大鼠使用改良线栓法制成I/R模型,原花青素低剂量组和原花青素高剂量组分别使用80 mg/kg和160 mg/kg的原花青素灌胃处理,连续处理1周。比较各组大鼠脑含水量;采用神经行为学评估比较各组大鼠神经功能缺损评分;使用试剂盒检测各组大鼠超氧化物歧化酶(SOD)、活性氧和谷胱甘肽过氧化物酶(GPX)水平;采用蛋白质印迹法检测TLR4/NF-κB信号通路和凋亡蛋白B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶3(Caspase-3)表达水平;采用苏木素染色观察神经细胞凋亡率。结果与对照组比较,模型组大鼠神经功能缺损评分、脑含水量、活性氧含量水平、TLR4蛋白、NF-κB蛋白、Bax蛋白和Caspase-3蛋白表达、神经细胞凋亡率升高,SOD含量水平、GPX含量水平、Bcl-2蛋白降低(P<0.05),其中TLR4蛋白和NF-κB蛋白分别为(1.80±0.20)和(1.85±0.21),显著低于对照组的(0.45±0.04)和(0.51±0.04);相比模型组,使用原花青素处理各组大鼠神经功能评分、脑含水量、活性氧含量水平、TLR4蛋白、NF-κB蛋白、Bax蛋白和Caspase-3蛋白表达、神经细胞凋亡率降低,且原花青素高剂量组低于原花青素低剂量组(P<0.05),其中原花青素高剂量组TLR4蛋白和NF-κB蛋白分别为(0.59±0.06)和(0.64±0.06),显著低于原花青素低剂量组的(1.00±0.10)和(0.98±0.09);与模型组比较,使用原花青素处理各组大鼠SOD含量水平、GPX含量水平、Bcl-2蛋白升高,且原花青素高剂量组高于原花青素低剂量组(P<0.05)。结论原花青素过抑制TLR4/NF-κB信号通路调控下游凋亡蛋白的表达Objective To explore protective effect of procyanidins(PRO)on cerebral ischemia-reperfusion(I/R)injury in rats by in⁃hibiting Toll-like receptor 4(TLR4)/NF-κB signaling pathway.Methods Sixty male SD rats were divided into control group(Control),model group(Model),low-dose PRO group(80 mg/kg)and high-dose PRO group(160 mg/kg)by random number table method,15 cases in each group.Except control group,modified suture method was applied in the other groups to prepare I/R models.The low-dose and high-dose PRO group were intragastrically administrated with 80 mg/kg and 160 mg/kg PRO,respectively,while control group and mod⁃el group were intragastrically administered with the same amount of normal saline,once/day for 1 week.The brain water content was compared among all groups.The nerve function deficits of each group were compared by neurobehavioral evaluation.Kit was applied to detect contents of superoxide dismutase(SOD),reactive oxygen species(ROS)and glutathione peroxidase(GPX)in each group.The ex⁃pression levels of TLR4/NF-κB signaling pathway related proteins and apoptosis proteins such as Bcl-2,Bax and Caspase-3 proteins in each group were detected by Western blotting.Hematoxylin staining was applied to observe apoptosis of nerve cells.Results Com⁃pared with control group,score of nerve function,water content in brain and ROS,expression of TLR4,NF-κB,Bax and Caspase-3 pro⁃teins,and apoptosis rate of nerve cells were increased in model group,levels of SOD and GPX,and Bcl-2 protein were decreased(P<0.05),among them,TLR4 protein and NF-κB protein were(1.80±0.20)and(1.85±0.21),respectively,which were significantly lower than those of the control group(0.45±0.04)and(0.51±0.04).Compared with model group,score of nerve function,water content in brain and ROS,expression of TLR4,NF-κB,Bax and Caspase-3 proteins,and apoptosis rate of nerve cells were decreased in all groups treat⁃ed with PRO,and the above indexes in high-dose PRO group were lower than those in low-dose PRO group(P<0.05),among them,the T

关 键 词：再灌注损伤 原花青素类 TOLL样受体4 核因子-ΚB 信号通路 细胞凋亡 大鼠 Sprague-Dawley 

分 类 号：R285.5[医药卫生—中药学]