食品中丙烯酰胺致小鼠睾丸生殖细胞凋亡的研究

为了解丙烯酰胺致小鼠生殖毒性的细胞凋亡机制,昆明种小鼠经丙烯酰胺(20、40、60mg/kg体重)慢性染毒8w后,应用末端转移标记技术(TUNEL)和流式细胞仪技术(Annexin-VFITC/PI)检测不同浓度丙烯酰胺对睾丸生殖细胞凋亡的影响。结果表明:两种方法检测结果均显示丙烯酰胺能引起睾丸生殖细胞的凋亡,而且随着浓度的增加细胞凋亡也逐渐增多,40、60mg/kg组的细胞凋亡率显著高于阴性对照(p<0.05),20mg/kg组的凋亡率虽然较阴性对照有所增加,但差异不显著;TUNEL法测定的细胞凋亡率高于Annexin-VFITC/PI,但两种方法测定结果无显著差异。可见,丙烯酰胺可以诱导小鼠睾丸生殖细胞发生凋亡,Annexin-VFITC/PI法及TUNEL法均能敏感检测出细胞凋亡,而且Annexin-VFITC/PI法具有更高的特异性。

Two Methods to Quantitate the Apoptosis Rate of Testes Cells from Mice after Acrylamide Exposure

To probe the apoptosis mechanism of reproductive toxicity to testes cells from mice after acrylamide exposure. Kunming mices were given acrylamide at dose of 20,40 and 60 mg/kg b w. by intraperitoneal injection for 8 weeks. Apoptosis rate was measured by Annexin-V FITC/PI and TUNEL. The results suggested that the apoptosis rate of 20,40 and 60 mg/kg groups detected by Annexin-V FITC/PI assay were 0.14±0.04,0.32±0.01 and 0.38±0.02 respectively. Those by TUNEL assay were 0.18±0.04,0.37±0.01 and 0.44±0.02 respectively. Both methods suggested that the apoptosis rate in 40 and 60mg/kg bw. groups were significantly higher than that of negative group(p<0.05). The results between two assays had no significantly differences(p>0.05). To concluded,acrylamide plays a crisis role to apoptosis of testes cells from mice and the key factor was the dose but not the exposure time to mice. Moreover,both of methods were qualified as a important role to quantitate the apoptosis rate. The Annexin-V FITC/PI assay was more specific.