玉米赤霉烯酮对小鼠原代培养T淋巴细胞细胞因子分泌的影响
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摘要
为探讨玉米赤霉烯酮(zearalenone,ZEA)对动物免疫功能的影响及其机理,试验以分离培养的原代小鼠脾脏淋巴细胞材料,研究了ZEA对T淋巴细胞细胞因子分泌的影响。以刀豆蛋白A(Con A,5 mg/L)作为T细胞活化特异性刺激剂,试验设空白对照组(不加Con A)、Con A组(5 mg/L Con A)、不同浓度的ZEA染毒组(Con A+ZEA 10、20和40μmol/L),处理48,72 h后,使用流式液相多重蛋白定量技术检测T细胞分泌IL-2、IL-3、IL-5、IL-6和GM-CSF等细胞因子的含量。结果显示,与空白对照组相比,Con A组细胞在48,72 h时间段5种细胞因子分泌均明显上升;与Con A组比较,染毒48,72 h后,10,20,40μmol/L ZEA染毒组各细胞因子分泌浓度均有极显著下降(P<0.01),并呈剂量-效应关系。结果表明,ZEA可以抑制T淋巴细胞细胞因子IL-2、IL-3、IL-5、IL-6和GM-CSF的分泌,从而影响机体正常免疫反应的进行,降低整体的免疫功能。
To investigate the effect and mechanism of zearalenone(ZEA) on immune function in mice,we isolated and cultured mouse primary spleen lymphocytes in vitro to study the effects of ZEA on the cytokine secretion of T lymphocytes.Concanavalin A(Con A,5 mg/L) was used as a specific T cell activation stimulator and the blank control group(without Con A),Con A group(5 mg/L) and three different concentrations of ZEA(10,20,40 μmol/L) were set.The levels of five cytokines IL-2,IL-3,IL-5,IL-6 and GM-CSF secreted by T lymphocytes were detected by Cytometric Bead Array at 48 h and 72 h respectively.The results showed that,compared with the blank control group,the secretion of 5 cytokines in the Con A group were significantly increased 48 h and 72 h.When the ZEA were added,the secretion of five cytokines decreased with the increase of the concentration of ZEA.Compared with Con A group,the secretion of cytokines in 10,20 and 40 μmol/L ZEA groups significantly decreased(P<0.01) at 48 h and 72 h,with a significant dose-effect relationship.The results showed that ZEA can inhibit the secretion of IL-2,IL-3,IL-5,IL-6 and GM-CSF of T lymphocytes,and thus affect the normal immune response and reduce the overall immune function.
To investigate the effect and mechanism of zearalenone(ZEA) on immune function in mice,we isolated and cultured mouse primary spleen lymphocytes in vitro to study the effects of ZEA on the cytokine secretion of T lymphocytes.Concanavalin A(Con A,5 mg/L) was used as a specific T cell activation stimulator and the blank control group(without Con A),Con A group(5 mg/L) and three different concentrations of ZEA(10,20,40 μmol/L) were set.The levels of five cytokines IL-2,IL-3,IL-5,IL-6 and GM-CSF secreted by T lymphocytes were detected by Cytometric Bead Array at 48 h and 72 h respectively.The results showed that,compared with the blank control group,the secretion of 5 cytokines in the Con A group were significantly increased 48 h and 72 h.When the ZEA were added,the secretion of five cytokines decreased with the increase of the concentration of ZEA.Compared with Con A group,the secretion of cytokines in 10,20 and 40 μmol/L ZEA groups significantly decreased(P<0.01) at 48 h and 72 h,with a significant dose-effect relationship.The results showed that ZEA can inhibit the secretion of IL-2,IL-3,IL-5,IL-6 and GM-CSF of T lymphocytes,and thus affect the normal immune response and reduce the overall immune function.
引文
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[2] YINGWANG,SHENGLILIU,HAOZHENG,et al.T-2 toxin,zearalenone and fumonisin B1 in feedstuffs from China [J].Food Addit Contam Part B Surveill,2013,6(2):116-22.
[3] HOU Y J,ZHU C C,XU Y X,et al.Zearalenone exposure affects mouse oocyte meiotic maturation and granulosa cell proliferation [J].Environ Toxicol,2015,30(10):1226.
[4] 何学军,齐德生.玉米赤霉烯酮的毒性研究进展 [J].中国饲料,2006(10):2-5.
[5] 杜宗亮.当前猪无名高热综合征的现状分析与防控对策 [J].黑龙江畜牧兽医,2007(1):63-65.
[6] YANG L,YANG W,FENG Q,et al.Effects of purified zearalenone on selected immunological measurements of blood in post-weaning gilts [J].Anim Nutr,2016,2(3):142-148.
[7] 梁梓森,马勇江,刘长永,等.玉米赤霉烯酮对小鼠免疫器官的毒性作用 [J].中国兽医科学,2010(3):279-283.
[8] SCHUMANN B,WINKLER J,MICKENAUTSCH N,et al.Effects of deoxynivalenol (DON),zearalenone (ZEN),and related metabolites on equine peripheral blood mononuclear cells (PBMC) in vitro and background occurrence of these toxins in horses [J].Mycotoxin Res,2016,32(3):153-161.
[9] 张娜,封颖璐.刀豆球蛋白A刺激对人外周血T淋巴细胞的增殖作用 [J].临床医学工程,2016,02):155-157.
[10] YIN S,MENG Q,ZHANG B,et al.Alleviation of zearalenone toxicity by modified halloysite nanotubes in the immune response of swine [J].Food Addit Contam Part A Chem Anal Control Expo Risk Assess,2015,32(1):87-99.
[11] 梁梓森,许利娜,马勇江,等.玉米赤霉烯酮对小鼠胸腺上皮细胞的毒性作用 [J].中国兽医学报,2009,29(7):894-897.
[12] 王亚超,邓俊良,徐世文.玉米赤霉烯酮对体外培养鸡脾脏淋巴细胞内环境稳态的影响[C]//中国畜牧兽医学会家畜内科学分会代表大会暨学术研讨会,2011.
[13] 潘顺叶,王光光,范文桐,等.玉米赤霉烯酮诱导CTLL-2细胞凋亡途径的研究 [J].中国兽医科学,2016(3):398-402.
[14] FULCHER D,WONG S.Carboxyfluorescein succinimidyl ester-based proliferative assays for assessment of T cell function in the diagnostic laboratory [J].Immunol Cell Biol,1999,77(77):559-564.
[15] 赵定亮,单风平.白细胞介素-2最新研究进展 [J].微生物学杂志,2013,33(4):77-83.
[16] MALEK T R.The biology of interleukin-2 [J].Annu Rev Immunol,2008,26:453-479.
[17] 张新海.白介素3与疾病的关系 [J].临床与病理杂志,1997,3):226-229.
[18] LUO X J,LI M,HUANG L,et al.The interleukin 3 gene (IL3) contributes to human brain volume variation by regulating proliferation and survival of neural progenitors [J].Plos One,2012,7(11):e50375.
[19] 高春芳,毕新岭,顾军.白介素5与相关皮肤病的研究进展 [J].国际皮肤性病学杂志,2002,28(4):239-42.
[20] KISHIMOTO T.The biology of interleukin-6 [J].Blood,1989,74(1):1.
[21] ISHIKAWA F.Use of rHuG-CSF for the treatment of myeloid leukemia and in targeting leukemia stem cells [J].Milestones Drug Therapy,2012,43:169-182.
[22] KING I L,KROENKE M A,SEGAL B M.GM-CSF-dependent,CD103+ dermal dendritic cells play a critical role in Th effector cell differentiation after subcutaneous immunization [J].J Exp Med,2010,207(5):953-961.