背景与目的:探讨巯基乙酸(thioglycolic acid,TGA)对小鼠卵母细胞胞浆成熟和相关生化指标的影响。材料与方法:以不同浓度TGA(0.2、1.0、2.5mmol/L)体外培养小鼠卵母细胞,并设M16培养液对照组,培养16h后在体视显微镜下,观测卵母细胞生发泡破裂和第一极体排出情况,用免疫荧光染色方法对皮质颗粒(cortical granules,CGs)进行标记,采用westem blot方法对p44/42MAPK进行检测。结果:各组生发泡破裂率均达到90%左右,各组间差异无统计学意义(P〉0.05)。随着TGA剂量的增加,第一极体排出率下降,各剂量组和对照组相比差异均有统计学意义(P〈0.05)。各剂量组均可观察到CGs在细胞膜下的线状排列,但随着TGA剂量的增加,质膜下CGs的密度降低,胞浆中CGs的密度有增加的趋势。对照组卵母细胞中有非常明显的无皮质颗粒区(cortical granules free domain,CGFD)形成,0.2mmol/L组也形成了CGFD,但1.0mmol/L和2.5mmol/L两个剂量组未观察到CGFD。1.0mmol/L和2.5mmol/L TGA可抑制p44/42MAPK的活化。结论:TGA可影响小鼠卵母细胞细胞质成熟和MAPK的活性,具有一定的生殖毒性。
BACKGROUND AND AIM: To explore the effect of Thioglycolic acid (TGA) on cytoplasmic maturation and related molecular factors of mouse oocytes. MATERIALS AND METHODS:Mouse oocytes were matured in vitro cultured with serial doses of TGA. Immunoflurescence staining was used to label cortical granules (CGs) and p44/ 42MAPK was measured by westem blot. RESULTS: Rate of germinal vesicle-breakdown in each group reached 90% and comparison was insignificant (P 〉 0.05). Rate of first polar body decreased with increasing TGA dose, the comparison between control and treatments was significant (P 〈 0.05).CGs of oocytes from all groups migrated to the cortex and formed a continuous layer under the cell membrane, but CGs density in the cytoplasm became higher with increasing TGA treatment dose. Obvious cortical granule free domain (CGFD) was observed at 0 mmol/L TGA, in 0.2 mmol/L TGA CGFD could still be seen, but not in 1.0 mmol/L and 2.5 mmol/L TGA groups. Meanwhile, TGA inhibited p44/42MAPK activation in 1.0 mmol/L and 2.5 mmol/L treatment groups. CONCLUSION: TGA exerted reproductive toxicity since it interfered with cytoplasmic maturation and MAPK activation of mouse oocytes.