三氯卡班对大鼠免疫毒性的初步研究  

Preliminary Study of Triclocarban on Immunotoxicity in Rats

作　　者：李芳芳 隋鑫 闵婷玉 黄涛[1] 徐在品[1] LI Fangfang;SUI Xin;MIN Tingyu;HUANG Tao;XU Zaipin(College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵阳550025

出　　处：《中国畜牧兽医》2024年第10期4626-4634,共9页China Animal Husbandry & Veterinary Medicine

基　　金：贵州省科技计划项目(黔科合支撑[2018]2265)。

摘　　要：【目的】研究三氯卡班(triclocarban,TCC)对大鼠免疫功能及脾脏Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子κB(NF-κB)信号通路相关因子表达的影响。【方法】试验选取96只5周龄雄性SD大鼠随机分为4组,分别为玉米油对照组(C组)以及0.1、20、100 mg/kg TCC组(TL、TM和TH组),每组24只。将不同剂量TCC与玉米油充分混匀,每日灌胃给药1次,C组灌胃等量不含TCC的玉米油,连续60 d。试验结束后,采集大鼠血液和脾脏组织,并检测大鼠血清生化指标、脾淋巴细胞增殖能力、巨噬细胞吞噬指数、血清溶血素水平以及脾脏自然杀伤细胞(natural killer cell,NK)活性。同时通过ELISA法检测大鼠血清免疫球蛋白含量;利用流式细胞术检测大鼠外周血淋巴细胞亚群水平;采用实时荧光定量PCR和Western blotting分别检测TLR4/MyD88/NF-κB通路标志基因以及下游因子的mRNA和蛋白表达水平。【结果】与C组相比,TH组大鼠血清中葡萄糖(Glu)水平显著降低(P<0.05),TM组大鼠血清肌酐(CREA)水平显著升高(P<0.05);TM组大鼠脾淋巴细胞增殖能力和血清溶血素水平均显著降低(P<0.05)。ELISA检测结果显示,TH组大鼠血清中免疫球蛋白A(IgA)、IgM、IgG水平及TM组大鼠血清中IgG水平与C组相比均极显著降低(P<0.01)。流式细胞检测结果显示,不同剂量TCC组大鼠外周血CD3^(+)、CD4^(+)T细胞占比和CD4^(+)/CD8^(+)比值与C组相比均无显著差异(P>0.05)。实时荧光定量PCR结果显示,与C组相比,不同剂量TCC组大鼠脾脏中TLR4、MyD88、IL-1β的mRNA相对表达量均显著或极显著升高(P<0.05;P<0.01)。Western blotting结果显示,与C组相比,TM组大鼠脾脏中TLR4、MyD88和NF-κB蛋白表达量均极显著升高(P<0.01)。【结论】TCC对大鼠的细胞免疫和体液免疫能够产生一定的毒性作用,能显著激活大鼠脾脏TLR4/MyD88/NF-κB信号通路,且可能通过靶向该信号通路来损伤大鼠的免疫系统,诱发大鼠的免疫毒【Objective】The purpose of this study was to investigate the effects of triclocarban(TCC)on immune function and the expression of Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factorκB(NF-κB)signaling pathway in spleen of rats.【Method】A total of 965-week-old male SD rats were randomly divided into 4 groups:Corn oil control group(group C)and 0.1,20 and 100 mg/kg TCC groups(TL,TM and TH),with 24 rats in each group.Different doses of TCC were thoroughly mixed with corn oil and given intragastric administration once a day.Rats in group C were given intragastric administration of the same amount of corn oil without TCC,the experiment lasted for 60 days.After the experiment,blood and spleen tissues of rats were collected,and serum biochemical indexes,splenic lymphocyte proliferation capacity,macrophage phagocytosis index,serum hemolysin level and spleen natural killer cell(NK)activity were detected.The serum immunoglobulin content of rats was detected by ELISA.The lymphocyte subsets in peripheral blood of rats were detected by flow cytometry.Real-time quantitative PCR and Western blotting were used to detect mRNA and protein expression levels of TLR4/MyD88/NF-κB pathway marker genes and downstream factors,respectively.【Result】Compared with group C,serum glucose(Glu)level in TH group was significantly decreased(P<0.05),and serum creatinine(CREA)level in TM group was significantly increased(P<0.05).The splenic lymphocyte proliferation ability and serum hemolysin level of TM group were significantly decreased(P<0.05).ELISA results showed that the levels of immunoglobulin A(IgA),IgM and IgG in serum of rats in TH group and IgG in serum in TM group were extremely significantly decreased compared with group C(P<0.01).Flow cytometry results showed that,compared with group C,there were no significant differences in the proportion of CD3^(+),CD4^(+)T cells and the ratio of CD4^(+)and CD8^(+)in peripheral blood of rats in different doses of TCC groups(P>0.05).Real-time quantitative PCR

关 键 词：三氯卡班 大鼠 Toll样受体4(TLR4) 髓样分化因子88(MyD88) 核因子κB(NF-κB) 免疫毒性 

分 类 号：S854.43[农业科学—临床兽医学]