邻苯二甲酸二(2-乙基)己酯对大鼠睾丸巨噬细胞凋亡的影响

Effect of DEHP on apoptosis of testicle macrophages among rats

目的研究邻苯二甲酸二(2-乙基)己酯(DEHP)对大鼠睾丸巨噬细胞(TM)凋亡的影响,为探讨免疫-内分泌途径在环境污染物雄性生殖毒性中的作用提供依据。方法采用剥离提取法原代培养大鼠TM,运用形态学观察、HE染色、墨汁吞噬实验及CD68抗原的免疫荧光实验鉴定细胞。分别用0(对照)、10、100、1 000μmol/L DEHP处理TM 24 h,采用流式细胞仪检测凋亡情况。结果原代培养72 h,贴壁的细胞呈圆形、梭形和不规则形,有突起,胞浆丰富,具有巨噬细胞的形态特征,细胞纯度大于90%。墨汁吞噬率>90%,提示细胞吞噬功能良好;CD68抗原鉴定为巨噬细胞,与对照组比较,各DEHP染毒组TM的凋亡率均较高,差异有统计学意义(P<0.05);且随着DEHP染毒剂量的升高,TM的凋亡率呈上升趋势。结论 DEHP能诱发大鼠TM细胞的凋亡。

Objective To investigate the effect of phthalate （2-ethylhexyl） ester （DEHP） on apoptosis of primary cultured rat＇s testicle macrophages （TMs）,which were useful for further studies on immune-endocrine role in the male reproductive toxicity of environmental pollutants. Methods Primary TMs were cultured by stripping extraction and identified by morphological observation, H.E dyeing,Chinese ink＇s phagocytosis and identification of CD68 immune fluorescent light. TMs were exposed to 0 （control）, 10, 100 and 1 000 μmol/L DEHP for 24 h respectively. TM apoptosis was detected by flow cytometry. Results Adherent TMs after 72 h were round,fusiform and irregularly shaped with projections and abundant cytoplasm, which showed morphology of macrophage. TM purity was greater than 90%. The rate of phagocytosis was greater than 90% indicating good function. The identification of CD68 immune fluorescent light proved to be macrophage. The rates of apoptosis in DEHP groups were significantly higher than that in control group （P〈0.05）. The rates of apoptosis obviously increased with the increase of DEHP. Conclusion DEHP may induce apoptosis of primary TMs.