补阳还五汤对一氧化碳中毒迟发性脑病大鼠氧化损伤及Nrf2/HO-1信号通路的影响  

Influence of Buyang Huanwu Tang on Oxidative Damage and Nrf2/HO-1 Signaling Pathway in Rats with Delayed Encephalophathy after Acute Carbon Monoxide Poisoning

作　　者：张谦[1] 赵俊喜[1] 景苗苗 符强 张小荣[1] ZHANG Qian;ZHAO Junxi;JING Miaomiao;FU Qiang;ZHANG Xiaorong(Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou 730050,China)

机构地区：[1]甘肃省中医院,甘肃兰州730050

出　　处：《西部中医药》2023年第3期35-40,共6页Western Journal of Traditional Chinese Medicine

基　　金：甘肃省自然科学基金(21JR11RA212)。

摘　　要：目的:探讨补阳还五汤对一氧化碳中毒迟发性脑病(delayed encephalophathy after acute carbon monoxide poisoning,DEACMP)大鼠氧化损伤的脑组织保护作用及其对核转录因子2(nuclear transcription factor 2,Nrf2)/血红素氧合酶1(heme oxygenose 1,HO-1)信号通路的影响。方法:采用改良式间隔腹腔注射法制作DEACMP大鼠模型;应用血气分析仪测定碳氧血红蛋白(carboxyhemaglobin,COHb)浓度;应用Morris水迷宫试验观察大鼠行为学表现;将造模成功的大鼠随机分为模型组(n=30)和补阳还五汤(BHD)高、中、低剂量组(n=30),以注射等量空气作为正常组(AC组);HE染色方法检测大鼠海马区病理变化;流式细胞术检测大鼠海马组织细胞凋亡情况;生化实验方法检测大鼠血液中丙二醛(malonaldehyde,MDA)、8-羟基脱氧鸟苷(8-hydroxy-2 deoxyguanosine,8-OHdG)水平;Western blot和qPCR方法检测大鼠海马组织中血红素氧合酶1(heme oxygenose 1,HO-1)、核因子红细胞系相关因子2(nuclear transcription factor 2,Nrf2)、Kelch样环氧氯丙烷相关蛋白1(Keap1)、磷酸化Nrf2(p-Nrf2)蛋白及其mRNA表达;蛋白质免疫共沉淀(Co-Immunoprecipitation,CoIP)方法检测Nrf2与Keap1的相互作用。结果:补阳还五汤可改善DEACMP模型大鼠海马组织损伤,减少炎症细胞浸润和组织水肿,降低海马组织细胞凋亡;提高海马组织HO-1、Nrf2、p-Nrf2蛋白及其mRNA表达,降低Keap1蛋白及其mRNA表达水平;提高海马组织细胞核中Nrf2蛋白与Keap1蛋白的相互作用量;降低大鼠血液中MDA和8-OHdG含量。结论:补阳还五汤可激活大鼠海马组织中Nrf2/HO-1信号通路,进而保护海马组织免遭急性一氧化碳中毒所致的氧化应激损伤,能抑制大鼠海马区神经元的持续凋亡,进而预防DEACMP。Objective:To survey the protective effects of Buyang Huanwu Tang on oxidative damage in DEACMP rats and its influence on Nrf2/HO-1 signaling pathway.Methods:DECAMP rat models were prepared by using modifying intraperitoneal injection;the concentrations of COHb were detected by applying blood gas analyzer;behavioral performance of the rats were observed by using Morris water maze;the rat models after successfully modeling were randomized into DEACMP group(n=30),high,moderate and low doses groups of Buyang Huanwu Tang(n=30),the control group was established by injecting the same amount of air(AC group);pathological changes in the hippocampus were measured by using HE staining;cellular apoptosis in the hippocampus were checked by adopting flow cytometry;the levels of MDA and 8-OHdG were detected in the blood by biochemical experimental methods;the expressions of HO-1,Nrf2,Keap1,p-Nrf2 protein and mRNA in the hippocampus were detected by Western blot and qPCR method;the interaction of Nrf2 and Keap1 were measured by colP method.Results:Buyang Huanwu Tang could improve the injury of hippocampal tissue in DEACMP rat models,reduce the infiltrating inflammatory cells and tissue edema,lower cellular apoptosis in hippocampal tissue;raise the expressions of HO-1,Nrf2,p-Nrf2 protein and mRNA in the hippocampus,lower the levels of Keap1 protein and mRNA;increase the interaction amount of Nrf2 protein and Keap1 protein in the nucleus of hippocampal tissue;reduce the contents of MDA and 8-OHdG in the blood of the rats.Conclusion:Buyang Huanwu Tang could promote the activation of Nrf2/HO-1 signaling pathway in hippocampal tissue,and then further prevent hippocampal tissue from oxidative stress damage induced by acute carbon monoxide poisoning,and it could inhibit the sustained apoptosis of neurons in the rat hippocampus,thereby preventing DEACMP.

关 键 词：迟发性脑病 一氧化碳中毒 Nrf2/HO-1信号通路 抗氧化 补阳还五汤 大鼠 

分 类 号：R259[医药卫生—中西医结合]