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[饮用水摄入三氯乙烯对BALB/c小鼠Th17细胞的影响]

[Effect of trichloroethylene intake via drinking water on Th17 cells in BALB/c mice].

作者信息

Shen Tong, Wang Jin, Xu Hui, Xu Shu-Hai, Jiang Tao, Zhu Qi-Xing

机构信息

Department of Toxicology, School of Public Health, Anhui Medical University, Hefei 230032, China. Email:

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 2012 Feb;46(2):152-7.

Abstract

OBJECTIVE

This study aimed to investigate the effect of trichloroethylene (TCE) intake via drinking water on Th17 cells in mice.

METHODS

Forty eight six weeks old female BALB/c mice were divided into blank control, vehicle control, 2.5 mg/ml TCE and 5.0 mg/ml TCE groups by random number table (12 mice each group), and exposed to TCE by drinking water. On the 14(th), 28(th), 56(th), 84(th) days, blood were collected and assayed for IL-17, IL-6, and TGF-β concentration in serum through ELISA. Animals were killed and spleen biopsies were taken sterility. The proportion of Th17 cells among CD4(+) T cells and RORγt mRNA expression level in spleen were measured by FCM and real-time PCR.

RESULTS

In 2.5 mg/ml TCE and 5.0 mg/ml TCE group mice, Th17 cells/CD4(+) T cells in spleen were (3.46 ± 0.32)% and (5.45 ± 0.45)% on day 14, (3.47 ± 0.33)% and (4.10 ± 0.39)% on day 84, which were significantly higher than those for solvent control group at the same time point ((2.15 ± 0.20)%, (2.16 ± 0.35)%, respectively) (P < 0.01). RORγt mRNA expression levels were (1.870 ± 0.084) and (1.965 ± 0.060) on 14 day, (1.998 ± 0.079) and (2.028 ± 0.073) on day 56, which were also significantly higher than those for solvent control group at the same time point (1.77 ± 0.04 and 1.75 ± 0.09, respectively) (P < 0.05). IL-17 concentrations in serum were (32.28 ± 5.38) and (34.47 ± 5.02) pg/ml on day 14, and (34.87 ± 5.48) and (41.94 ± 6.19) pg/ml on day 28, which were significantly higher than those for solvent control group at the same time point((21.57 ± 5.23), (22.11 ± 5.11) pg/ml). IL-6 concentration in serum were (43.07 ± 6.71) and (47.86 ± 8.52) pg/ml on day14, (41.32 ± 7.04) and (46.74 ± 9.33) pg/ml on day 56, which were significantly higher than solvent control group at the same time point ((7.56 ± 7.71) and (28.26 ± 7.22) pg/ml). TGF-β concentration were (17.48 ± 3.06) and (18.93 ± 3.12) pg/ml on day 14, which did not show significant difference from solvent control group ((15.25 ± 2.95) pg/ml). Correlation analysis showed that IL-6 in serum were significantly positively correlated with the proportion of Th17 cells among CD4(+) T cells and RORγt expression level in spleen (r = 0.741, 0.765, P < 0.01).

CONCLUSION

TCE might promote the differentiation of Th17 cells and increase IL-17 secretion by inducing IL-6 and up-regulating RORγt expression together with TGF-β.

摘要

目的

本研究旨在探讨经饮水摄入三氯乙烯(TCE)对小鼠Th17细胞的影响。

方法

将48只6周龄雌性BALB/c小鼠通过随机数字表法分为空白对照组、溶剂对照组、2.5 mg/ml TCE组和5.0 mg/ml TCE组(每组12只),通过饮水方式使其接触TCE。在第14、28、56、84天采集血液,采用酶联免疫吸附测定法(ELISA)检测血清中白细胞介素-17(IL-17)、白细胞介素-6(IL-6)和转化生长因子-β(TGF-β)的浓度。处死动物,无菌采集脾脏组织。采用流式细胞术(FCM)和实时荧光定量聚合酶链反应(real-time PCR)检测脾脏中CD4(+) T细胞中Th17细胞的比例以及维甲酸相关孤儿受体γt(RORγt)mRNA表达水平。

结果

在2.5 mg/ml TCE组和5.0 mg/ml TCE组小鼠中,第14天时脾脏中Th17细胞/CD4(+) T细胞比例分别为(3.46 ± 0.32)%和(5.45 ± 0.45)%,第84天时分别为(3.47 ± 0.33)%和(4.10 ± 0.39)%,均显著高于同一时间点的溶剂对照组(分别为(2.15 ± 0.20)%、(2.16 ± 0.35)%)(P < 0.01)。第14天时RORγt mRNA表达水平分别为(1.870 ± 0.084)和(1.965 ± 0.060),第56天时分别为(1.998 ± 0.079)和(2.028 ± 0.073),也显著高于同一时间点的溶剂对照组(分别为1.77 ± 0.04和1.75 ± 0.09)(P < 0.05)。第14天时血清中IL-17浓度分别为(32.28 ± 5.38)和(34.47 ± 5.02) pg/ml,第28天时分别为(34.87 ± 5.48)和(41.94 ± 6.19) pg/ml,均显著高于同一时间点的溶剂对照组((21.57 ± 5.23)、(22.11 ± 5.11) pg/ml)。第14天时血清中IL-6浓度分别为(43.07 ± 6.71)和(47.86 ± 8.52) pg/ml,第56天时分别为(41.32 ± 7.04)和(46.74 ± 9.33) pg/ml,均显著高于同一时间点的溶剂对照组((7.56 ± 7.71)和(28.26 ± 7.

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