百草枯对小鼠脾脏淋巴细胞的影响
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摘要
[背景]百草枯(PQ)是一种广泛使用的除草剂。目前的研究主要针对PQ的神经毒性。然而,PQ对外周免疫系统影响的研究尚不全面。[目的 ]观察PQ对外周免疫系统的影响,特别是对淋巴细胞数量、分化及活化状态的影响。[方法 ] 6～8周龄SPF级雌性C57BL/6小鼠随机分为对照组、2 mg/kg PQ染毒1周组和2周组,每组8只小鼠。采用皮下注射方式染毒,染毒频率为每周一次,分别于染毒1、2周后取脾脏,用流式细胞术检测脾脏细胞和淋巴细胞数量,以及淋巴细胞分化和活化情况。[结果 ]与对照组相比,PQ染毒小鼠脾脏细胞的数量在第1周和第2周差异无统计学意义(P> 0.05);CD4+T细胞和CD8+T细胞的数量在第1周和第2周差异亦无统计学意义(P>0.05);B细胞的数量在第2周时高于对照组(t=-2.304,P <0.05),在第1周时差异无统计学意义(P> 0.05)。与对照组相比,PQ在第2周时降低辅助性T细胞1(Th1细胞)(t=0.019,P <0.05)和辅助性T细胞2(Th2细胞)(t=0.038,P <0.05)占CD4+T细胞的比例,在第1周时无影响(P> 0.05);不影响辅助性T细胞17(Th17细胞)和调节性T细胞(Treg细胞)占CD4+T细胞的比例(P> 0.05)。此外,PQ在第2周时降低B细胞表面主要组织相容性复合体Ⅱ类分子I-A(t=0.047,P <0.05)和B细胞表面抗原CD40(t=0.000,P <0.01)的表达,不影响B细胞表面抗原CD86的表达(P> 0.05)。[结论 ] PQ可增加B细胞数量,可以抑制CD4+T细胞向Th1和Th2方向分化,并抑制B细胞活化,提示PQ可以通过影响免疫细胞数量、分化和活化而引起免疫功能紊乱。
[Background] Paraquat(PQ) is a widely used herbicide. Current research focuses on its neurotoxicity. However, the impact of PQ on peripheral immune system is not comprehensively studied.[Objective] This study aims to study the effects of PQ on peripheral immune system, especially on the number, differentiation, and activation of lymphocytes.[Methods] SPF female C57 BL/6 mice, 6-8 weeks old, were randomly assigned to three groups,namely control group, 2 mg/kg PQ for 1 week group, and for 2 weeks group, with 8 mice in each group. Mice were subcutaneously injected with PQ once a week. Spleen was harvested 1 or 2 weeks after exposure to measure the number of splenocytes and lymphocytes, as well as the differentiation and activation of lymphocytes by flow cytometry.[Results] Compared with the control group, the number of splenocytes did not change after 1 or 2 weeks of PQ exposure(P > 0.05); the numbers of CD4+T cells and CD8+ T cells did not change either(P > 0.05); the number of B cells was higher after 2 weeks of exposure(t=-2.304, P < 0.05),but not after 1 week of exposure(P > 0.05). Compared with the control group, PQ decreased the proportions of type 1 T helper(Th1) cells(t=0.019, P < 0.05) and type 2 T helper(Th2) cells(t=0.038,P < 0.05) to CD4+T cells after 2 weeks of exposure, but not after 1 week of exposure(P > 0.05); PQ did not affect the proportions of T helper 17(Th17) cells and regulatory T(Treg) cells(P > 0.05)to CD4+T cells. In addition, PQ reduced the expressions of a major histocompatibility complex class Ⅱ molecule I-A(t=0.047, P < 0.05) and B-cell surface antigen CD40(t=0.000, P < 0.01) after 2 weeks of exposure, but did not affect the expression of B-cell surface antigen CD86(P > 0.05).[Conclusion] PQ exposure could increase the number of B cells, inhibit the differentiation of CD4+T cells into Th1 and Th2 cells, and inhibit the activation of B cells, suggesting that PQ could cause immune dysfunction by affecting the number, differentiation, and activation of immune cells.
[Background] Paraquat(PQ) is a widely used herbicide. Current research focuses on its neurotoxicity. However, the impact of PQ on peripheral immune system is not comprehensively studied.[Objective] This study aims to study the effects of PQ on peripheral immune system, especially on the number, differentiation, and activation of lymphocytes.[Methods] SPF female C57 BL/6 mice, 6-8 weeks old, were randomly assigned to three groups,namely control group, 2 mg/kg PQ for 1 week group, and for 2 weeks group, with 8 mice in each group. Mice were subcutaneously injected with PQ once a week. Spleen was harvested 1 or 2 weeks after exposure to measure the number of splenocytes and lymphocytes, as well as the differentiation and activation of lymphocytes by flow cytometry.[Results] Compared with the control group, the number of splenocytes did not change after 1 or 2 weeks of PQ exposure(P > 0.05); the numbers of CD4+T cells and CD8+ T cells did not change either(P > 0.05); the number of B cells was higher after 2 weeks of exposure(t=-2.304, P < 0.05),but not after 1 week of exposure(P > 0.05). Compared with the control group, PQ decreased the proportions of type 1 T helper(Th1) cells(t=0.019, P < 0.05) and type 2 T helper(Th2) cells(t=0.038,P < 0.05) to CD4+T cells after 2 weeks of exposure, but not after 1 week of exposure(P > 0.05); PQ did not affect the proportions of T helper 17(Th17) cells and regulatory T(Treg) cells(P > 0.05)to CD4+T cells. In addition, PQ reduced the expressions of a major histocompatibility complex class Ⅱ molecule I-A(t=0.047, P < 0.05) and B-cell surface antigen CD40(t=0.000, P < 0.01) after 2 weeks of exposure, but did not affect the expression of B-cell surface antigen CD86(P > 0.05).[Conclusion] PQ exposure could increase the number of B cells, inhibit the differentiation of CD4+T cells into Th1 and Th2 cells, and inhibit the activation of B cells, suggesting that PQ could cause immune dysfunction by affecting the number, differentiation, and activation of immune cells.
引文
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[14]窦婷婷,常秀丽,吕文,等.百草枯对人胚胎神经干细胞分化过程中Wnt通路分子表达的影响[J].环境与职业医学,2013,30(6):411-415.
[15]李强,刘海英,王玉红,等.百草枯中毒患者免疫功能改变及临床意义[J].新乡医学院学报,2006,23(6):575-576.
[16]蒋白丽,李建平,李文倩,等.百草枯中毒患者外周血淋巴细胞亚群变化及对预后判断的作用[J].中国工业医学杂志,2018,31(2):114-115.
[17]王海燕.白芍总苷辅助治疗对过敏性紫癜患儿辅助性T细胞、高迁移率族蛋白1表达的影响[J].中医药信息,2019,36(1):76-80.
[18]刘环,陈嵘祎,张方,等.IL-36β联合CpG-DNA对B细胞增殖、活化及TLR9表达的影响[J].广东医科大学学报,2018,36(5):505-510.
[19]HENN V,SLUPSKY J R,GR?FE M,et al.CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells[J].Nature,1998,391(6667):591-594.
[20]WIECZOREK M,ABUALROUS E T,STICHT J,et al.Major histocompatibility complex(MHC)class I and MHC class IIproteins:conformational plasticity in antigen presentation[J].Front Immunol,2017,8:292.
[2]ZHAO F,WANG W,WANG C,et al.Mfn2 protects dopaminergic neurons exposed to paraquat both in vitro and in vivo:implications for idiopathic Parkinson's disease[J].Biochim Biophys Acta Mol Basis Dis,2017,1863(6):1359-1370.
[3]ZHANG X F,THOMPSON M,XU Y H.Multifactorial theory applied to the neurotoxicity of paraquat and paraquatinduced mechanisms of developing Parkinson's disease[J].Lab Invest,2016,96(5):496-507.
[4]RIAHI B,RAFATPANAH H,MAHMOUDI M,et al.Immunotoxicity of paraquat after subacute exposure to mice[J].Food Chem Toxicol,2010,48(6):1627-1631.
[5]RIAHI B,RAFATPANAH H,MAHMOUDI M,et al.Evaluation of suppressive effects of paraquat on innate immunity in Balb/c mice[J].J Immunotoxicol,2011,8(1):39-45.
[6]HASSUNEH M R,ALBINI M A,TALIB W H.Immunotoxicity induced by acute subtoxic doses of paraquat herbicide:implication of shifting cytokine gene expression toward T-helper(TH)-17 phenotype[J].Chem Res Toxicol,2012,25(10):2112-2116.
[7]LIM J H,WON J H,AHN K H,et al.Paraquat reduces natural killer cell activity via metallothionein induction[J].JImmunotoxicol,2015,12(4):342-349.
[8]HSIEH C S,MACATONIA S E,TRIPP C S,et al.Pillars article:development of TH1 CD4+T cells through IL-12 produced by Listeria-induced macrophages.1993.Science 260(5107):547-549[J].J Immunol,2008,181(7):4437-4439.
[9]WANG J,MA L,YANG S,et al.IL-10-expressing Th2cells contribute to the elevated antibody production in rheumatoid arthritis[J].Inflammation,2016,39(3):1017-1024.
[10]SHAO S,YU X,SHEN L.Autoimmune thyroid diseases and Th17/Treg lymphocytes[J].Life Sci,2018,192:160-165.
[11]AN N,CHEN Y,WANG C,et al.Chloroquine autophagic inhibition rebalances Th17/Treg-mediated immunity and ameliorates systemic lupus erythematosus[J].Cell Physiol Biochem,2017,44(1):412-422.
[12]NAKAGAWA I,SUZUKI M,IMURA N,et al.Involvement of oxidative stress in paraquat-induced metallothionein synthesis under glutathione depletion[J].Free Radic Biol Med,1998,24(9):1390-1395.
[13]杨正丽,张玉彬,周志俊.百草枯的神经免疫毒性研究进展[J].环境与职业医学,2017,34(11):1009-1012.
[14]窦婷婷,常秀丽,吕文,等.百草枯对人胚胎神经干细胞分化过程中Wnt通路分子表达的影响[J].环境与职业医学,2013,30(6):411-415.
[15]李强,刘海英,王玉红,等.百草枯中毒患者免疫功能改变及临床意义[J].新乡医学院学报,2006,23(6):575-576.
[16]蒋白丽,李建平,李文倩,等.百草枯中毒患者外周血淋巴细胞亚群变化及对预后判断的作用[J].中国工业医学杂志,2018,31(2):114-115.
[17]王海燕.白芍总苷辅助治疗对过敏性紫癜患儿辅助性T细胞、高迁移率族蛋白1表达的影响[J].中医药信息,2019,36(1):76-80.
[18]刘环,陈嵘祎,张方,等.IL-36β联合CpG-DNA对B细胞增殖、活化及TLR9表达的影响[J].广东医科大学学报,2018,36(5):505-510.
[19]HENN V,SLUPSKY J R,GR?FE M,et al.CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells[J].Nature,1998,391(6667):591-594.
[20]WIECZOREK M,ABUALROUS E T,STICHT J,et al.Major histocompatibility complex(MHC)class I and MHC class IIproteins:conformational plasticity in antigen presentation[J].Front Immunol,2017,8:292.