摘要
目的探讨脂氧素A4(LXA4)对脓毒症大鼠脑损伤的作用以及对Toll样受体4(TLR4)/NADPH氧化酶4(NOX4)/核转录因子-κB(NF-κB)信号通路的影响。方法随机将大鼠分为4组(n=20):对照组(S组):仅进行开关腹处理;脓毒症模型组(CLP组):进行盲肠结扎穿孔处理;脂氧素A4治疗组(LXA4组):进行盲肠结扎穿刺后予脂氧素A410μg/kg腹腔注射;TLR4受体激动剂干预组Pam3Cys(Pam组):进行盲肠结扎穿刺后予LXA450μg/kg和2 mg/kg的Pam3Cys腹腔注射;同时造模成功后24 h,对大鼠进行神经功能评分和脑组织含水量检测,采集大鼠下腔静脉血液和脑组织。采用苏木精伊红(HE)观察脑组织病理变化,酶联免疫吸附试验(ELISA)检测肿瘤坏死因子α(TNF-α)、白介素1(IL-1)、白介素6(IL-6)和髓过氧化物酶活性(MPO),通过定量逆转录聚合酶链反应(q-RT-PCR)和蛋白质印迹(western blot)检测脑组织中TLR4、NOX4和NF-κB信号通路mRNA和蛋白的表达量。结果与S组相比,其余3组脑组织病理显示脑损伤改变,神经功能评分和脑组织含水量增高,TNF-α、IL-1、IL-6,MPO均显著升高,TLR4、NOX4和NF-κB信号通路mRNA和蛋白的表达量升高(P<0.05);与CLP组和Pam组相比,LXA4组的脑组织病理损伤减轻,神经功能评分和脑组织含水量降低,TNFα、IL-6、MPO显著下降,TLR4、NOX4和NF-κB信号通路基因和蛋白的表达量明显降低(P<0.05),CLP组和Pam组结果之间无显著差异(P>0.05)。结论脂氧素A4可通过抑制TLR4/NOX4/NF-κB通路,降低TNFα、IL-1、IL-6,MPO水平,抑制氧化应激反应从而减轻脓毒症引起的脑损伤。
Objective To investigate the effects of lipoxin A4(LXA4)on brain injury in rats with sepsis and on Toll-like receptor 4(TLR4)/NADPH oxidase 4(NOX4)/nuclear transcription factor-kappa B(NF-κB)signaling pathway.Methods A total of 80 rats were divided into four groups according to random number table method,with 20 rats in each group.The rats in control group were treated by opening and closing abdomen only,the rats in CLP group were treated by cecum ligation and perforation,and the rats in LXA4 group were treated by peritoneal injection with Lipoxin A410μg/kg after cecal ligation and puncture,and the rats in Pam group were treated by peritoneal injection with LXA450μg/kg and Pam3Cys 2mg/kg after cecal ligation and puncture.At 24 hours after the successful modeling,the neural function score and brain tissue water content were measured,and the blood specimens and brain tissue of the rats were collected.Hematoxylin eosin staining(HE staining)was used to observe the pathological changes of brain tissue.Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factorα(TNF-α),interleukin 1(IL-1),interleukin 6(IL-6)and myeloperoxidase activity(MPO).Quantitative reverse transcription polymerase chain reaction(q-RT-PCR)and Western Blot were used to detect the expression levels of TLR4,NOX4 and NF-κB signaling pathway mRNA and protein in brain tissues.Results As compared with control group,the other three groups showed the changes of brain tissue injury,and the neurological function scores and water content in brain tissue were significantly increased,and the levels of TNF-α,IL-1,IL-6,MPO,and the expression levels of TLR4,NOX4 and NF-κB signaling pathway mRNA and protein were significantly increased(P<0.05).As compared with CLP group and Pam group,LXA4 group showed that the pathological damage of brain tissue were relieved,and neurological function scores,water content of brain tissue,the levels of TNF-α,IL-6,MPO and the expression levels of TLR4,NOX4 and NF-κB signaling pathway genes
作者
朱国辉
魏锋
ZHU Guohui;WEI Feng(Department of ICU,Chifeng College Affiliated Hospita of Chifeng College,Inner Mongolia Autonomous Region,Chifeng 024000,China)
出处
《河北医药》
CAS
2021年第8期1156-1160,共5页
Hebei Medical Journal
基金
内蒙古自治区高等学校科学研究项目(编号:NJZY16249)。
