围生期低剂量4-壬基酚暴露对亲代Wistar雌性大鼠的免疫毒性研究  

Study on the immunotoxicity of low-dose 4-nonylphenol exposure during perinatal period on parental Wistar female rats

作　　者：谢静华 闫九明[1] 程树品 杨溢 王贺 陈锦瑶[1] 张立实[1] XIE Jing-hua;YAN Jiu-ming;CHENG Shu-pin;YANG Yi;WANG He;CHEN Jin-yao;ZHANG Li-shi(West China School of Public Health and West China Fourth Hospital,Sichuan University,Chengdu Sichuan 610041,China)

机构地区：[1]四川大学华西公共卫生学院/四川大学华西第四医院,四川成都610041

出　　处：《毒理学杂志》2022年第1期43-48,54,共7页Journal of Toxicology

基　　金：国家自然科学基金青年科学基金(81703258)。

摘　　要：目的探讨围生期低剂量4-壬基酚(4-NP)暴露对亲代雌性Wistar大鼠的免疫毒性及相关作用机制。方法按照受孕时体重将母鼠分为空白对照组和低、中和高剂量组,从妊娠(GD)第6天至产后(PND)第21天分别给予正常和4-壬基酚拌饲量依次为10、100和500 mg/kg的NIH-31改良饲料。PND21后,将母鼠随机分为两部分:一部分进行空斑实验和血清凝血素测定;另一部分进行NK细胞活性测定和脾淋巴细胞转化试验,并测定血清细胞因子含量。此外,每组随机抽取3只母鼠测量外周血和脾淋巴细胞亚群以及脾中叉头状转录因子3(Foxp3)和孤独核受体γt(RORγt)mRNA表达量、磷酸化信号转导和转录激活因子3(p-STAT3)、磷酸化信号转导和转录激活因子5(p-STAT5)和磷酸化酪氨酸激酶3(p-JAK3)的蛋白水平。结果与空白组相比,高剂量组GD21和GD22母鼠体重明显降低(P<0.01),高剂量组PND4、PND7、PND11体重明显低于空白组(P<0.05);胸腺和脾高剂量组脏器系数明显低于空白组(P<0.05);高剂量组血清凝集程度和空斑数明显低于空白组(P<0.05),同时3个剂量组的NK细胞活性均明显降低(P<0.05);外周血中高剂量组B细胞占白细胞的比例高于空白组(P<0.05),外周血和脾中、高剂量组辅助性T细胞17(Th17)/调节性T细胞(Treg)比值均明显高于空白组(P<0.01);与空白组相比,高剂量组血清转化生长因子-β(TGF-β)、白细胞介素-17A(IL-17A)含量及3个剂量组血清白细胞介素-10(IL-10)含量、TGF-β/IL-17A比值均明显降低(P<0.05);各剂量组脾Foxp3/RORγt mRNA比值均低于空白组(P<0.05);低和中剂量组脾p-STAT3、p-STAT5及高剂量组p-JAK3蛋白水平明显升高(P<0.05)。结论围生期低剂量4-壬基酚暴露对亲代雌性大鼠具有一定的免疫毒性,其可能机制与4-NP可通过激活JAK-STAT信号通路,导致p-STAT3和p-STAT5持续激活,诱导RORγt表达,促进Th17细胞分化和IL-17的释放,使Th17/Treg平衡左移有关。Objective To explore the immunotoxicity and related mechanism of perinatal low-dose 4-nonylphenol exposure on parental female Wistar rats.Methods Wistar females were divided into blank control group and low-dose,medium-dose and high-dose groups according to their weight at conception.From GD6 to PND21,the NIH-31 modified diets were supplemented with 0,10,100 and 500 mg/kg 4-nonylphenol,respectively.After PND21,the female rats were randomly divided into two parts,half of which were tested for plaque and serum thrombin.The other half were tested for NK cell activity and splenic lymphocyte transformation test,and serum cytokine content was determined.In addition,3 female rats were randomly selected from each group to measure the lymphocyte subsets in peripheral blood and spleen,as well as the mRNA expression levels of Forkhead box P3(FoxP3)and Orphan Nuclear ReceptorγT(RORγT)in spleen Protein levels of phosphorylated signal transduction and transcriptional activator 3(p-STAT3),phosphorylated signal transduction and transcriptional activator 5(p-STAT5)and phosphorylated tyrosine kinase 3(p-JAK3).Results Compared with blank group,the body weight of GD21 and GD22 in high-dose group was significantly decreased(P<0.01),and the body weight of PND4,PND7 and PND11 in high-dose group was significantly lower than that in blank group(P<0.05).The organ coefficient of thymus and spleen high-dose group was significantly lower than that of blank group(P<0.05).Compared with blank group,serum agglutination degree and plaque number in high dose group were significantly decreased(P<0.05),and NK cell activity in three dose groups were significantly decreased(P<0.05).The proportion of B cells to WBC in peripheral blood of high dose group was higher than that of blank group(P<0.05),and the ratio of helper T cells 17(Th17)/regulatory T cells(Treg)in peripheral blood,spleen and medium and high dose groups was significantly higher than that of blank group(P<0.01).Compared with blank group,the contents of transforming growth factor-β(TGF-

关 键 词：壬基酚 免疫毒性 围生期 亲代大鼠 低剂量暴露 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]