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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 investigate the effect of MEHP and cadmium chloride on the secretion of interleukin-10 (IL-10) by rat testicular macrophages. Methods Testicular macrophages (TM) of SPF adult male Wistar rats were isolated and purified by adherence method and exposed to 0, 1, 10, 10, 100 and 1000 μmol / L MEHP Or cadmium chloride solution for 24 h, the cell viability was determined by CCK8 assay. The cells were exposed to a final concentration of 0 (control, 0.1% DMSO) and 1, 10, 100μmol / L of MEHP and / or 0.1,1,10μmol / ml LPS). After cultured for 24 h, the secretion of IL-10 by cells was detected by ELISA. Results Compared with the control group, the inhibitory rates of TM cells in 100, 1000μmol / L MEHP or 10,100,1 000μmol / L cadmium chloride exposure group were higher than those in control group (P <0.05) With the increase of MEHP or cadmium chloride exposure, the inhibitory rate of TM cells in rats showed an upward trend. After activation with 5μg / ml LPS, the secretion of cytokine IL-10 in each exposure group was higher than that in the control group, except for 100μmol / L MEHP + 10μmol / L cadmium chloride exposure group, the differences were statistically significant (P <0.05 ). Compared with the activator group, the concentrations of MEHP and cadmium chloride, alone or in combination, had a higher inhibitory rate on IL-10 secreted by rat TM (P <0.05), and with MEHP and ( Or) cadmium chloride exposure, the inhibitory rate of IL-10 secreted by rat TM all showed an upward trend. Compared with the MEHP plus cadmium chloride exposure group, the inhibition rates of IL-10 secretion in MEHP exposed group and 1,10 μmol / L cadmium chloride exposure group were lower than those in MEHP plus cadmium chloride exposure group Significance (P <0.05). Factorial analysis showed synergistic effects of 10 μmol / L MEHP + 1 μmol / L cadmium chloride exposure and 100 μmol / L MEHP + 10 μmol / L cadmium chloride exposure on IL-10 secretion by rat TM. Conclusions Under the experimental conditions, the synergistic effect of MEHP and cadmium chloride exposure on IL-10 secretion in rat TM cells is synergistic.