摘要
目的通过检测急性暴露脱氧雪腐镰刀菌烯醇(DON)后小鼠半数死亡数(LD50)值、生长指数,血液、血生化指标变化以及病理观察,探讨小鼠急性DON染毒的毒性效应,并通过检测染毒小鼠的淋巴细胞分群等指标,关注小鼠染毒DON对免疫系统的影响。方法选取清洁级昆明小鼠50只,20只用于上下法测定小鼠LD50值,另30只随机分为对照组、低剂量组(2 mg/kg·bw)、高剂量组(10 mg/kg·bw),每组10只,灌胃给予DON 1次,恢复喂养14 d后取材,观察指标为血液与血生化、主要脏器(心、肺、胸腺、脾、肾和肝)称重、脏体比与组织病理学观察,包括苏木素-伊红(HE)染色观察、Tunel法观察细胞凋亡,免疫组化观察细胞凋亡相关蛋白P53、Bax、bcl-2蛋白表达以及外周血淋巴细胞分型。结果上下法测定昆明小鼠DON的LD50值为17.5 mg/kg·bw。与对照组比较,低剂量组各指标无明显差别(P>0.05);高剂量组小鼠部分血小板(PLT)增高、淋巴细胞(LYM)降低,血生化指标中天冬氨酸转氨酶(ALT)、丙氨酸转氨酶(AST)、乳酸脱氢酶(LDH)、碱性磷酸酶(ALP)明显下降,总胆红素(TBIL)、白蛋白(ALB)、肌酐(CREA)显著升高(P<0.05);高剂量组小鼠肝、肾、脾相对重量与对照组相比有统计学差异(P<0.05);Tunel实验可见高剂量组小鼠肝、肾、脾、胸腺出现凋亡细胞增多,同时高剂量组小鼠脾NK细胞比值高于对照组,B细胞低于对照组(P<0.05)。结论本试验条件下雄性小鼠急性经口毒性LD50为17.5 mg/kg·bw,一次急性染毒DON可以诱导肝、肾、脾、胸腺的细胞凋亡,造成小鼠的免疫毒性,其作用机制可能在于脱氧雪腐镰刀菌烯醇活化p53信号通路。
Objective The acute oral toxicities of DON in male mice was investigated by detecting LD50,growth index,blood and serum biochemical markers and pathological changes in mice after acute exposure to deoxynivalenol（DON）. Moreover,by detecting the lymphocyte grouping in blood,including the ratio of T lymphocytes,B lymphocytes and NK cells,the effects of DON on the immune system in mice were focused on. Methods The LD50 value of DON was determined by Up-and-Down procedure（UDP） with 20 male mice. Based on LD50,30 male mice were randomly divided into control group,low dose group（2. 0 mg/kg·bw） and high dose group（10. 0 mg/kg·bw）,10 rats in each group,and these mice were gavaged with water or DON one time. After feeding 14 days,indicators of hematology,biochemistry,body weight and the weight of main organs（heart, lung, thymus, heart, kidney, spleen） weight were observed.Histopathologically examination of main organs was examined by haematoxylin-eosin staining. The apoptosis in different organs were observed by Tunel method. The expressions of P53,Bax,bcl-2 proteins were measured by immunohistochemistry. The ratios of T lymphocytes,B lymphocytes and NK cells in blood were detected by flow cytometer. Results The LD50 value of DON is 17. 5 mg/kg·bw by Up-and-Down procedure. Compared with the control group,there was no significant difference in low dose group（P〈0. 05）. PLT increased and LYM decreased significantly in hematology parameters at high dose group（P〈0. 05）. In biochemistry,the decreases of ALT,AST,LDH,Urea and the increase of TBIL,ALB,CREA,CHE were observed in high dose group（P〈0. 05）. Compared with the control group,the increases of relative weight of liver,kidney,and spleen were observed in high dose group（P〈0. 05）. For lymphocyte,the change was shown by the increase of NK cell and decrease of B cell in lymphocytes in blood by Tunel assay（P〈0. 05）. Conclusion In the present study,The LD50 value of DON is 17. 5 mg/kg·bw in male mice,the acute oral exposure of DON could induce the apoptosis of the liver,kidney,spleen and thymus,and resulted in immunotoxicity in mice,the underlying mechanism might be due to the activation of p53 signal pathway by deoxynivalenol.
作者
支媛
方业鑫
刘海波
崔文明
冯晓莲
汪会玲
ZHI Yuan1, FANG Ye-xin2 , LIU Hai-bo1 , CUI Wen-ming1 , FENG Xiao-lian1 , WANG Hui-ling1(1. China National Center for Food Safety Risk Assessment, Beijing 100022, China; 2. Anhui Province Anqing No. 2 Middle School, Anqing Anhui 200463, Chin)
出处
《毒理学杂志》
CAS
CSCD
2018年第1期11-17,共7页
Journal of Toxicology
基金
国家自然科学基金(81302462)
北京市自然科学基金(7142128)
国家食品安全风险评估中心高层次人才队伍建设523项目
