邻苯二甲酸单乙基己酯和镉对大鼠睾丸巨噬细胞分泌白介素-10的影响

Effects of MEHP and cadmium on IL-10 in testicular macrophages in rats

目的研究邻苯二甲酸单乙基己酯(MEHP)及氯化镉对大鼠睾丸巨噬细胞分泌细胞因子白介素-10(IL-10)的影响。方法利用贴壁法提取SPF级成年雄性Wistar大鼠的睾丸巨噬细胞(TM),分别暴露于终浓度0(对照,0.1%DMSO)、0.1、1、10、100、1 000μmol/L的MEHP或氯化镉溶液培养24 h,采用CCK8法测定细胞活性。并将细胞分别暴露于终浓度0(对照,0.1%DMSO)及1、10、100μmol/L的MEHP和(或)0.1、1、10μmol/L的氯化镉溶液,并加入激活剂(5μg/ml LPS),培养24 h后,采用ELISA法检测细胞分泌IL-10的情况。结果与对照组比较,100、1 000μmol/L MEHP或10、100、1 000μmol/L氯化镉暴露组大鼠TM细胞抑制率均较高,差异有统计学意义(P<0.05);且随着MEHP或氯化镉暴露浓度的升高,大鼠TM细胞抑制率均呈上升趋势。经5μg/ml LPS激活后,各暴露组细胞因子IL-10的分泌量高于对照组,除100μmol/L MEHP+10μmol/L氯化镉暴露组外,差异均有统计学意义(P<0.05)。与激活剂组比较,各浓度MEHP和氯化镉单独或联合暴露对大鼠TM分泌IL-10的抑制率均较高,差异均有统计学意义(P<0.05);且随着MEHP和(或)氯化镉暴露浓度的升高,对大鼠TM分泌IL-10的抑制率均呈上升趋势。与相同浓度MEHP+氯化镉联合暴露组比较,10、100μmol/L的MEHP暴露组及1、10μmol/L氯化镉暴露组大鼠TM细胞IL-10的分泌抑制率较低,差异均有统计学意义(P<0.05)。析因分析结果显示,10μmol/L MEHP+1μmol/L氯化镉暴露和100μmol/L MEHP+10μmol/L氯化镉暴露对大鼠TM分泌IL-10的抑制作用表现出协同作用。结论在本实验条件下,MEHP和氯化镉联合暴露对大鼠TM细胞IL-10的分泌抑制作用表现为协同作用。

Objective To study the effect of mono（2-ethylhexyl）phthalate（MEHP） and cadmium chloride on IL-10 cytokine secreted in rat testicular macrophages（TM）. Methods Adherence culture was used to extract TM of Wistar rat. With 0.1%DMSO as the control group and 5 μg/ml lipopoly-saccharide（LPS）as the activator,the cells were exposed to different doses（0.1,1,10,100,1 000 μmol/L） of MEHP or cadmium chloride respectively and their mixtures in a certain proportion for 24 hours. IL-10 cytokine secreted in the cells were determined by ELISA. Cell viability was detected by colorimetric method with CCK8 kit after different treatments. Results Compared with the control group,MEHP（100,1 000 μmol/L） and cadmium chloride（10,100,1 000 μmol/L） declined TM cell viability significantly（P〈0.05）,with dose dependent trend. 5 μg/ml LPS could obviously promote the TM to secrete IL-10 cytokine in every exposure group except 100 μmol/L MEHP ＋10 μmol/L cadmium chloride group. Compared with the activator group,single or joint exposure of MEHP and cadmium chloride at each concentration could decrease the contents of IL-10 cytokine obviously（P〈0.05）,with dose dependent trend. The inhibition rate of IL-10 cytokine secretion at MEHP（10,100 μmol/L） and cadmium chloride（1,10 μmol/L） group were less than that of same concentration joint exposure group significantly（P〈0.05）. Synergistic effect between MEHP and cadmium chloride was showed at 10 μmol/L MEHP＋1 μmol/L cadmium chloride and 100 μmol/L MEHP＋10 μmol/L cadmium chloride group on secretion of IL-10 in rat TM. Conclusion The inhibition of MEHP and cadmium chloride on secretion of IL-10 in TM has certain synergistic effect at moderate and higher dose groups under this experimental condition.