核转录因子-κB抑制剂二硫代氨基甲酸吡咯烷对急性一氧化碳中毒大鼠模型中iNOS动态变化的影响  

Effect of nuclear factor⁃κB inhibitor PDTC on the dynamic changes of iNOS in rats with acute carbon monoxide poisoning

作　　者：蒋力 雷蕊绮 彭红艳 辜刚凤 李经伦 JIANG Li;LEI Ruiqi;PENG Hongyan;GU Gangfeng;LI Jinglun(Department of Neurolo⁃gy,the Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院神经内科,四川泸州646000

出　　处：《实用医学杂志》2020年第10期1311-1317,共7页The Journal of Practical Medicine

基　　金：四川省泸州市科技计划项目(编号:2017LZXNYD⁃J36)。

摘　　要：目的通过使用二硫代氨基甲酸吡咯烷(pyrrolidine dithiocarbamate,PDTC)对核转录因子-κB的抑制,观察急性一氧化碳中毒大鼠中NF⁃κB、iNOS因子的动态变化,探讨急性一氧化碳迟发性脑病中的发病机制。方法120只成年健康雄性SD大鼠随机分为空气对照组(AC组)、一氧化碳中毒组(CO组)、一氧化碳中毒+PDTC干预组(PD组),按各组大鼠染毒后1、3、7、14、21 d时间点分为5个小组,采用Morris水迷宫检测大鼠平均逃避潜伏期、穿越平台次数、平台运动总时间的变化,评估大鼠学习记忆能力损害程度,Western Blot和免疫荧光检测大鼠海马区NF⁃κB、iNOS的表达,HE染色观察海马区细胞形态变化,TUNEL染色检测大鼠海马CA1区细胞凋亡情况。结果AC组仅可检测到少量NF⁃κB、iNOS蛋白的表达;CO组NF⁃κB表达呈现逐渐增多,其iNOS蛋白表达于第3天达到高峰后逐渐减少。PD组与CO组相比,都呈现出PDTC抑制NF⁃κB的活化后,NF⁃κB、iNOS蛋白表达相对减少,且组间各时间点表达差异具有统计学意义(P<0.05),PD组与AC组相比差异明显,差异亦具有统计学意义(P<0.05)。CO组、PD组大鼠凋亡细胞于第7天开始明显增多,PD组与CO组各时间点比较大鼠凋亡细胞相对减少,且差异有统计学意义(P<0.05)。结论NF⁃κB/iNOS信号通路参与了DEACMP的发病过程,PDTC的干预通过下调NF⁃κB的激活及减少iNOS的表达,从而减轻大鼠CO中毒症状。Objective To observe the dynamic changes of NF⁃κB and iNOS factors in rats with acute carbon monoxide poisoning by using PDTC(dithiocarbamate)to inhibit the nuclear factor⁃κB,and to explore the pathogenesis of acute carbon monoxide delayed encephalopathy.Methods A total of 120 adult healthy male SD rats were randomly divided into air control group(AC group),carbon monoxide poisoning group(CO group)and carbon monoxide poisoning+PDTC intervention group(PD group).The rats in each group were divided into 5 groups according to 1,3,7,14 and 21 days after exposure.Morris water maze was used to detect changes in mean escape latency,number of platform crossings and total platform movement time in rats,in order to evaluate the degree of learning and memory impairment.The expression of NF⁃κB,iNOS in hippocampus of rats was detected by Western Blot and Immunofluorescence assay;the morphological changes of hippocampal cells were observed by HE staining,and the apoptosis of hippocampal CA1 cells was detected by TUNEL staining.Results Only a small amount of NF⁃κB,iNOS protein expression was detected in AC group.The expression of NF⁃κB in CO group increased gradually,and the expression of iNOS protein decreased gradually after reaching the peak on the 3rd day.Compared with CO group,the expression of NF⁃κB,iNOS protein in PD group decreased relatively after PDTC inhibited the activation of NF⁃κB,and the expression of NF⁃κB,iNOS protein in PD group was significantly different from that in AC group at each time point(P<0.05).The apoptotic cells in CO group and PD group began to increase significantly on the 7th day,and the apoptotic cells in PD group were significantly lower than those in CO group at each time point,and the difference was statistically significant(P<0.05).Conclusion NF⁃κB/iNOS signaling pathway is involved in the pathogenesis of DEACMP.PDTC could relieve the symptoms of CO poisoning in rats by down⁃regulating the activation of NF⁃kB and reducing the expression of iNOS.

关 键 词：一氧化碳中毒 核转录因子-ΚB 诱导型NOS(iNOS) 一氧化氮 二硫代氨基甲酸吡咯烷(PDTC) 

分 类 号：R845[医药卫生—航空、航天与航海医学]