砷暴露对小鼠脾脏炎症反应及Th1/Th2的影响
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摘要
目的探讨慢性饮水砷暴露对小鼠脾脏的炎症反应及Th1/Th2细胞因子和转录因子表达的影响。方法将30只健康SPF级雌性昆明小鼠根据体重按照随机数字表分为对照组和25、50 mg/L NaAsO_2染毒组,每组10只。采用自由饮水方式染毒,染毒6个月后,测定小鼠脾脏组织中炎症因子IL-12和IL-6的mRNA水平。采用Real-time PCR方法分别检测脾脏Th1和Th2的转录因子T-bet、Gata3和细胞因子Ifn-γ、IL-4的mRNA表达水平。同时用Western blot法观察Nrf2、NQO1和GSTO1/2的蛋白表达水平。结果与对照组比较,随着染毒剂量的上升,各染毒组小鼠脾脏炎症因子IL-12和IL-6的mRNA水平均上升,除IL-12的25 mg/L剂量组外,差异均具有统计学意义(P<0.05);各染毒组小鼠脾脏细胞因子Ifn-γ和IL-4的转录水平呈上升趋势;Nrf2及其调控的下游蛋白NQO1和GSTO1/2的表达水平明显增加。结论慢性砷暴露小鼠免疫器官脾脏的炎症相关因子IL-12和IL-6明显增加,同时上调Th1和Th2特异性细胞因子的表达水平,调节CD4~+T淋巴细胞亚群的分化,从而发挥对小鼠脾脏的免疫调节效应。
Objective To investigate the effects of arsenic exposure by chronic drinking water on inflammatory response and function of Th1/Th2 in spleens of mice.Methods Thirty healthy SPF female Kunming mice were devided into 3 groups according to weight based on random number table: control group, 25 mg/L NaAsO_2 group and 50 mg/L NaAsO_2 group, 10 mice for each group.The mRNA levels of inflammatory cytokines IL-12 and IL-6 in spleens of mice were determined 6 months later after exposure to As by free drinking, and real-time PCR was used to detect the mRNA expression levels of Th1 and Th2 transcription factors such as T-bet, Gata3 and cytokines Ifn-γ and IL-4 in spleen, meanwhile, Western blot was used to observe the protein expression levels such as Nrf2, NQO1 and GSTO1/2.Results The results showed that with the increase of dose, the mRNA levels of inflammatory cytokines such as IL-12 and IL-6 in spleens of mice increased compared with control group(P<0.05), aside from IL-12 in 25 mg/L dose group; the transcription levels of cytokines Ifn-γ and IL-4 in each As-treated group also showed an upward trend, and the protein levels of Nrf2 and its regulated downstream proteins NQO1 and GSTO1/2 were significantly increased as well.Conclusion The results suggested that chronic arsenic exposure might up-regulate the inflammation related factors IL-12 and IL-6 of immune organ in mice, elevate the expression level of specific cytokines of Th1 and Th2 regulate the differentiation of CD4~+T lymphocyte subset, thereby played the immunoregulatory effect on spleen of mice.
Objective To investigate the effects of arsenic exposure by chronic drinking water on inflammatory response and function of Th1/Th2 in spleens of mice.Methods Thirty healthy SPF female Kunming mice were devided into 3 groups according to weight based on random number table: control group, 25 mg/L NaAsO_2 group and 50 mg/L NaAsO_2 group, 10 mice for each group.The mRNA levels of inflammatory cytokines IL-12 and IL-6 in spleens of mice were determined 6 months later after exposure to As by free drinking, and real-time PCR was used to detect the mRNA expression levels of Th1 and Th2 transcription factors such as T-bet, Gata3 and cytokines Ifn-γ and IL-4 in spleen, meanwhile, Western blot was used to observe the protein expression levels such as Nrf2, NQO1 and GSTO1/2.Results The results showed that with the increase of dose, the mRNA levels of inflammatory cytokines such as IL-12 and IL-6 in spleens of mice increased compared with control group(P<0.05), aside from IL-12 in 25 mg/L dose group; the transcription levels of cytokines Ifn-γ and IL-4 in each As-treated group also showed an upward trend, and the protein levels of Nrf2 and its regulated downstream proteins NQO1 and GSTO1/2 were significantly increased as well.Conclusion The results suggested that chronic arsenic exposure might up-regulate the inflammation related factors IL-12 and IL-6 of immune organ in mice, elevate the expression level of specific cytokines of Th1 and Th2 regulate the differentiation of CD4~+T lymphocyte subset, thereby played the immunoregulatory effect on spleen of mice.
引文
[1]Yu NH,Pei H,Huang YP,et al.(-)-Epigallocatechin-3-Gallate inhibits arsenic-induced inflammation and apoptosis through suppression of oxidative stress in mice[J].Cell Physiol Biochem,2017,41(5):1788-1800.
[2]Ferrario D,Gribaldo L,Hartung T.Arsenic exposure and immunotoxicity:a review including the possible influence of age and sex[J].Curr Environ Health Rep,2016,3(1):1-12.
[3]Biswas R,Ghosh P,Banerjee N,et al.Analysis of T-cell proliferation and cytokine secretion in the individuals exposed to arsenic[J].Hum Exp Toxicol,2008,27(5):381-386.
[4]Hernández-Castro B,Doníz-Padilla LM,Salgado-Bustamante M,et al.Effect of arsenic on regulatory T cells[J].J Clin Immunol,2009,29(4):461-469.
[5]Singh MK,Yadav SS,Yadav RS,et al.Efficacy of crude extract of Emblica officinalis (amla) in arsenic-induced oxidative damage and apoptosis in splenocytes of mice[J].Toxicol Int,2014,21(1):8-17.
[6]Soto-Peňa GA,Vega L.Arsenic interferes with the signaling transduction pathway of T cell receptor activation by increasing basal and induced phosphorylation of Lck and Fyn in spleen cells[J].Toxicol Appl Pharmacol,2008,230(2):216-226.
[7]Sankar P,Telang AG,Suresh S,et al.Immunomodulatory effects of nanocurcumin in arsenic-exposed rats[J].Int Immunopharmacol,2013,17(1):65-70.
[8]Ren X,McHale CM,Skibola CF,et al.An emerging role for epigenetic dysregulation in arsenic toxicity and carcinogenesis[J].Environ Health Perspect,2011,119(1):11-19.
[9]Singh MK,Yadav SS,Yadav RS,et al.Protective effect of emblica-officinalis in arsenic induced biochemical alteration and inflammation in mice[J].Springerplus,2015,4(1):438.
[10]Luo JH,Qiu ZQ,Shu WQ,et al.Effects of arsenic exposure from drinking water on spatial memory,ultra-structures and NMDAR gene expression of hippocampus in rats[J].Toxicol Lett,2009,184(2):121-125.
[11]Singh N,Kumar D,Lal K,et al.Adverse health effects due to arsenic exposure:modification by dietary supplementation of jaggery in mice[J].Toxicol Appl Pharmacol,2010,242(3):247-255.
[12]Prasad P,Sinha D.Low-level arsenic causes chronic inflammation and suppresses expression of phagocytic receptors[J].Environ Sci Pollut Res Int,2017,24(12):11708-11721.
[13]Zhang K,Zhao P,Guo G,et al.Arsenic trioxide attenuates NF-κB and cytokine mRNA levels in the livers of cocks[J].Biol Trace Elem Res,2016,170(2):432-437.
[14]Zhao H,He Y,Li S,et al.Subchronic arsenism-induced oxidative stress and inflammation contribute to apoptosis through mitochondrial and death receptor dependent pathways in chicken immune organs[J].Oncotarget,2017,8(25):40327-40344.
[15]Thomas-Schoemann A,Batteux F,Mongaret C,et al.Arsenic trioxide exerts antitumor activity through regulatory T cell depletion mediated by oxidative stress in a murine model of colon cancer[J].J Immunol,2012,189(11):5171-5177.
[16]Wang Y,Zhao H,Shao Y,et al.Copper or/and arsenic induce oxidative stress-cascaded,nuclear factor kappa B-dependent inflammation and immune imbalance,trigging heat shock response in the kidney of chicken[J].Oncotarget,2017,8(58):98103-98116.
[17]Lau A,Whitman SA,Jaramillo MC,et al.Arsenic-mediated activation of the Nrf2-Keap1 antioxidant pathway[J].J Biochem Mol Toxicol,2013,27(2):99-105.
[2]Ferrario D,Gribaldo L,Hartung T.Arsenic exposure and immunotoxicity:a review including the possible influence of age and sex[J].Curr Environ Health Rep,2016,3(1):1-12.
[3]Biswas R,Ghosh P,Banerjee N,et al.Analysis of T-cell proliferation and cytokine secretion in the individuals exposed to arsenic[J].Hum Exp Toxicol,2008,27(5):381-386.
[4]Hernández-Castro B,Doníz-Padilla LM,Salgado-Bustamante M,et al.Effect of arsenic on regulatory T cells[J].J Clin Immunol,2009,29(4):461-469.
[5]Singh MK,Yadav SS,Yadav RS,et al.Efficacy of crude extract of Emblica officinalis (amla) in arsenic-induced oxidative damage and apoptosis in splenocytes of mice[J].Toxicol Int,2014,21(1):8-17.
[6]Soto-Peňa GA,Vega L.Arsenic interferes with the signaling transduction pathway of T cell receptor activation by increasing basal and induced phosphorylation of Lck and Fyn in spleen cells[J].Toxicol Appl Pharmacol,2008,230(2):216-226.
[7]Sankar P,Telang AG,Suresh S,et al.Immunomodulatory effects of nanocurcumin in arsenic-exposed rats[J].Int Immunopharmacol,2013,17(1):65-70.
[8]Ren X,McHale CM,Skibola CF,et al.An emerging role for epigenetic dysregulation in arsenic toxicity and carcinogenesis[J].Environ Health Perspect,2011,119(1):11-19.
[9]Singh MK,Yadav SS,Yadav RS,et al.Protective effect of emblica-officinalis in arsenic induced biochemical alteration and inflammation in mice[J].Springerplus,2015,4(1):438.
[10]Luo JH,Qiu ZQ,Shu WQ,et al.Effects of arsenic exposure from drinking water on spatial memory,ultra-structures and NMDAR gene expression of hippocampus in rats[J].Toxicol Lett,2009,184(2):121-125.
[11]Singh N,Kumar D,Lal K,et al.Adverse health effects due to arsenic exposure:modification by dietary supplementation of jaggery in mice[J].Toxicol Appl Pharmacol,2010,242(3):247-255.
[12]Prasad P,Sinha D.Low-level arsenic causes chronic inflammation and suppresses expression of phagocytic receptors[J].Environ Sci Pollut Res Int,2017,24(12):11708-11721.
[13]Zhang K,Zhao P,Guo G,et al.Arsenic trioxide attenuates NF-κB and cytokine mRNA levels in the livers of cocks[J].Biol Trace Elem Res,2016,170(2):432-437.
[14]Zhao H,He Y,Li S,et al.Subchronic arsenism-induced oxidative stress and inflammation contribute to apoptosis through mitochondrial and death receptor dependent pathways in chicken immune organs[J].Oncotarget,2017,8(25):40327-40344.
[15]Thomas-Schoemann A,Batteux F,Mongaret C,et al.Arsenic trioxide exerts antitumor activity through regulatory T cell depletion mediated by oxidative stress in a murine model of colon cancer[J].J Immunol,2012,189(11):5171-5177.
[16]Wang Y,Zhao H,Shao Y,et al.Copper or/and arsenic induce oxidative stress-cascaded,nuclear factor kappa B-dependent inflammation and immune imbalance,trigging heat shock response in the kidney of chicken[J].Oncotarget,2017,8(58):98103-98116.
[17]Lau A,Whitman SA,Jaramillo MC,et al.Arsenic-mediated activation of the Nrf2-Keap1 antioxidant pathway[J].J Biochem Mol Toxicol,2013,27(2):99-105.