磷酸三苯酯对幼年小鼠血液和胸腺中免疫细胞及细胞因子的影响
Effects of triphenyl phosphate on immune cells and cytokines in blood and thymus of young mice
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摘要:
背景 磷酸三苯酯（TPHP）作为有机磷阻燃剂已广泛运用于多个行业。目前的研究主要针对TPHP的内分泌、生殖毒性，尚无免疫毒性的系统性研究。
目的 探究幼年小鼠TPHP暴露后血液及胸腺免疫细胞和细胞因子的变化。
方法 将28只幼年C57BL/6小鼠（4周）随机分为4组，包括对照组、50 mg·kg-1 TPHP组（低剂量组）、150mg·kg-1 TPHP组（中剂量组）和450mg·kg-1 TPHP组（高剂量组），每组7只。各组幼鼠连续灌胃28 d，每天1次，记录体重，收集血液，进行脏器称重并计算脏器系数，用全血细胞分析仪进行白细胞（WBC）计数，流式细胞术检测血液中淋巴细胞和髓系细胞以及胸腺中淋巴细胞的数目和比例；采用液相蛋白芯片技术检测血浆中的粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白介素（IL）-4、IL-1β、IL-17水平。
结果 TPHP染毒13d时高剂量组体重较对照组减少了11%（
P < 0.05），染毒28d后减少了12.6%（P < 0.01）。各剂量组的肝脏脏器系数均高于对照组（P < 0.05），但脾脏、胸腺和肾脏的脏器系数没有统计学差异（P >0.05）。低、中、高剂量组的血液WBC计数分别为（6.53±1.06）×106 mL-1、（6.91±1.50）×106 mL-1、（7.30±2.00）×106 mL-1，均较对照组（3.61±0.51）×106 mL-1升高（P < 0.01）。各剂量组血液中CD4+T细胞、CD8+T细胞及B细胞数目均较对照组增加（P < 0.01），但CD4+T细胞比例下降（P < 0.01）；胸腺WBC数目及CD4+T细胞、CD8+T细胞及CD4+CD8+T细胞数目和比例与对照组的差异均无统计学意义（P >0.05）。仅高剂量组血液中髓系细胞比例较对照组增加了29%（P < 0.05），低、中、高剂量组的髓系细胞数目较对照组分别增加了79%、82%、165%（P < 0.01），并且高剂量组血浆中IL-4水平（0.29±0.12）ng·L-1较对照组（0.14±0.04）ng·L-1升高了107%（P < 0.05）。结论 TPHP染毒可使幼鼠血液中的白细胞、淋巴细胞数目增加，并促进IL-4的分泌，诱导血液免疫紊乱。
Abstract:Background As an organophosphate flame retardant, triphenyl phosphate (TPHP) has been widely used in many industries. Current research mainly focuses on the endocrine and reproductive toxicities of TPHP, but there is limited information on its immunotoxicity.
Objective This study explores the changes of immune cells in blood and thymus and cytokines in young mice exposed to TPHP.
Methods Twenty-eight young C57BL/6 mice (4 weeks old) were randomly divided into four groups, including control group, 50 mg·kg-1 TPHP group (low dose group), 150 mg·kg-1 TPHP group (middle dose group), and 450 mg·kg-1 TPHP group (high dose group), with seven mice in each group. The young mice in each group were administered by gavage for consecutively 28 days, once a day, and the body weight was recorded. The blood and organs were collected to weigh and calculate organ coefficients. White blood cell (WBC) count was measured with blood analyser; counts and ratios of lymphocytes and myeloid cells in blood and of lymphocytes in thymus were detected by flow cytometry. The levels of granulocyte-macrophage colonystimulating factor (GM-CSF), interleukin (IL)-4, IL-1β, and IL-17 in plasma were measured by liquid protein chip technology.
Results After 13 days of TPHP exposure, the body weight of the high dose group was reduced by 11% compared with the control group (
P < 0.05), and it was reduced by 12.6% after 28 days (P < 0.01). The organ coefficients of liver in all dose groups were higher than that in the control group (P < 0.05), but there were no differences in the organ coefficients of spleen, thymus, and kidney between the exposed groups and the control group (P >0.05). The WBC counts in blood in the low, middle, and high dose groups were (6.53±1.06)×106 mL-1, (6.91±1.50)×106 mL-1, and (7.30±2.00)×106 mL-1, respectively, and all were higher than that in the control group(3.61±0.51)×106 mL-1 (P < 0.01). The numbers of CD4+T cells, CD8+T cells, B cells in blood in each dose group were higher than those in the control group (P < 0.01), but the ratio of CD4+T cells in each dose group was lower than that in the control group (P < 0.01). The number of thymus WBC and the numbers and proportions of thymus CD4+T cells, CD8+T cells, and CD4+CD8+T cells in each dose group were not different from those in the control group (P >0.05). Only the ratio of myeloid cells in blood in the high dose group was increased by 29% compared with the control group (P < 0.05). The numbers of myeloid cells in the low, middle and high dose groups were increased by 79%, 82%, and 165% respectively compared with the control group (P < 0.01), and the plasma IL-4 level in the high dose group(0.29±0.12) ng·L-1 was increased by 107% compared with the control group(0.14±0.04) ng·L-1 (P < 0.05).Conclusion TPHP exposure could increase the number of white blood cells and lymphocytes, and promote the secretion of IL-4, potentially inducing blood immune disorder.
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