邻苯二甲酸二丁酯和苯并(a)芘染毒对大鼠T淋巴细胞亚群比例及Foxp3表达水平的影响  

Effects of dibutyl phthalate and benzo(a)pyrene on the proportion of T lymphocytes and the expression of Foxp3 in rats

作　　者：李冬琴 陈静[1] 聂雪 罗密 王敏[1] 李宽 杨光红 LI Dongqin;CHEN Jing;NIE Xue;LUO Mi;WANG Min;LI Kuan;YANG Guanghong(School of Public Health,the Key Laboratory of Environmental Pollution Monitoring and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang 550025,China;Guizhou Provincial Center for Disease Control and Prevention,Guiyang 550004,China)

机构地区：[1]贵州医科大学公共卫生与健康学院,环境污染与疾病监控教育部重点实验室,贵州贵阳550025 [2]贵州省疾病预防控制中心中心办公室,贵州贵阳550004

出　　处：《社区医学杂志》2023年第9期448-455,共8页Journal Of Community Medicine

基　　金：国家自然科学基金(81760578);2019年国家级大学生创新创业训练计划项目(201910660038)。

摘　　要：目的探讨分析邻苯二甲酸二丁酯(DBP)和苯并(a)芘(BaP)的免疫损伤机制,通过DBP和BaP亚慢性染毒大鼠,分析T淋巴细胞亚群比例以及叉头螺旋翼状转录因子3(Foxp3)表达水平的变化情况,为DBP、BaP所致的免疫损伤机制研究及预防提供研究线索。方法将80只健康的SD大鼠随机分为空白对照组、溶剂对照组(给予玉米油)、DBP 50 mg/kg组(标为DBP_(50)组)、DBP 250 mg/kg组(标为DBP_(250)组)、BaP 1 mg/kg组(标为BaP_(1)组)、BaP 5 mg/kg组(标为BaP_(5)组)、DBP 50 mg/kg+BaP 1 mg/kg组(标为DBP_(50)+BaP_(1)组)、DBP 250 mg/kg+BaP 5 mg/kg组(标为DBP_(250)+BaP_(5)组),每组10只大鼠,采取灌胃方式进行染毒,连续染毒90 d,其中每周连续染毒6 d。染毒结束后提取及纯化大鼠淋巴细胞,采用流式细胞术检测淋巴细胞内各亚群比例;反转录实时荧光聚合酶链式反应法(RT-qPCR)检测Foxp3 mRNA的表达水平;蛋白质免疫印迹(WB)检测淋巴细胞内Foxp3蛋白的表达水平;实验结果用x±s表示,组间比较用方差分析,进行两两比较,若方差齐性采用Tukey’s检验,若方差不齐性采用Games-Howell检验,并采用Pearson相关分析Treg淋巴细胞比例与Foxp3蛋白表达水平、CD4^(+)T、CD8^(+)T淋巴细胞比例的相关性。结果DBP、BaP染毒后,CD4^(+)T淋巴细胞比例溶剂对照组(18.433±1.718)%与DBP_(50)组(6.873±0.645)%、DBP_(250)组(3.747±0.153)%、BaP_(1)组(6.303±0.746)%、BaP_(5)组(6.257±0.471)%、DBP_(50)+BaP_(1)组(6.933±0.628)%、DBP_(250)+BaP_(5)组(3.637±0.137)%比较,差异有统计学意义,P值分别为0.018、0.019、0.013、0.020、0.019和0.019;BaP_(5)组(6.257±0.471)%与DBP_(250)+BaP_(5)组(3.637±0.137)%比较,差异有统计学意义,F=194.80,P=0.034。CD8^(+)T淋巴细胞比例溶剂对照组(32.870±1.035)%与DBP_(50)组(29.703±0.319)%、DBP_(250)组(18.160±1.185)%、BaP_(1)组(30.707±1.320)%、BaP_(5)组(19.393±0.833)%、DBP_(50)+BaP_(1)组(30.247±1.665)%、DBP_(250)+BaP_(5)组(11.397±1.108)%比较,差�Objective To investigate the changes of T lymphocytes ratio and the expression of the forkhead/winged helix transcription factor 3(Foxp3) in rats exposed to dibutyl phthalate(DBP) and benzo(a)pyrene(BaP),to provide clues for the mechanism of DBP and BaP-induced immune injury.Methods Totally 80 SD rats purchased from Changsha Tianqin Biotechnology Co, Ltd were randomly divided into eight groups, including blank control group, solvent control group(corn oil),DBP_(50) mg/kg group(DBP_(50)),DBP_(250) mg/kg group(DBP_(250)),BaP_(1) mg/kg group(BaP_(1)),BaP_(5) mg/kg group(BaP_(5)),DBP_(50) mg/kg+BaP_(1) mg/kg group(DBP_(50)+BaP_(1)),and DBP_(250) mg/kg+BaP_(5) mg/kg group(DBP_(250)+BaP_(5)),with 10 rats in each group.The drug was administered by gavage for 90 days, with continuous exposure for 6 days a week.After exposure for 90 days, the lymphocytes of rats were extracted and purified, and the changes in the proportion of lymphocytes were detected by Flow cytometry;In addition, reverse transcription-real-time polymerase chain reaction(RT-qPCR) and western blot(WB) were used to detect the mRNA and protein expressions of Foxp3.Pearson correlation was used to analyze the correlation between the proportion of Treg lymphocytes and the protein expression of Foxp3,and the proportion of CD4^(+)T and CD8^(+)T lymphocytes.Results After DBP and BAP dyeing, the solvent control group of CD4^(+)T lymphocytes(18.433±1.718) % compared with DBP_(50) group(6.873±0.645)%,DBP_(250) group(3.747±0.153)%,BAP1 group(6.303±0.746)%,BaP_(5) group(6.257±0.471)%,DBP_(50)+BaP_(1) group(6.933±0.628)%,DBP_(250)+BaP_(5) group(3.637±0.137)%,the difference was statistically significant, the P-value were respectively 0.018,0.019,0.013,0.02,0.019 and 0.019;BaP_(5) group(6.257±0.471) % compared with DBP_(250)+BaP_(5) group(3.637±0.137) %,the difference was statistically significant, F=194.80,P=0.034.The solvent control group of CD8^(+)T lymphocytes(32.870±1.035)% compared with DBP_(50) group(29.703±0.319)%,DBP_(250) group(18.160±1.185)%,BaP_(

关 键 词：邻苯二甲酸二丁酯 苯并(A)芘 淋巴细胞 叉头螺旋翼状转录因子3 

分 类 号：R322.2[医药卫生—人体解剖和组织胚胎学] R992[医药卫生—基础医学]