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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 DEHP on the apoptosis of rat testis macrophages (TM) and to provide a theoretical basis for exploring the role of the immune-endocrine pathway in male reproductive toxicity of environmental pollutants in accordance with. Methods Primary cultured rat TM was isolated by stripping method, and the cells were identified by morphological observation, HE staining, ink ingestion phagocytosis and immunofluorescence assay of CD68 antigen. TM was treated with 0 (control), 10,100,1 000μmol / L DEHP treatment 24 h, the apoptosis was detected by flow cytometry. Results Primary culture 72 h, adherent cells were round, fusiform and irregular shape, with protrusions, abundant cytoplasm, macrophages morphological features, cell purity greater than 90%. The phagocytosis rate of ink was> 90%, which indicated that phagocytosis of cells was good. CD68 antigen was identified as macrophages. Compared with the control group, the apoptosis rate of TM was higher in all DEHP groups (P <0.05) ; And with the increase of DEHP dose, the apoptosis rate of TM increased. Conclusion DEHP can induce the apoptosis of rat TM cells.