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目的研究天花粉蛋白(TCS)对小鼠卵母细胞第1次减数分裂的影响。方法性成熟昆明雌鼠,注射孕马血清促性腺激素10IU后48h,从卵巢中获取未成熟卵母细胞共480枚,在含不同浓度TCS的培养液中,分组进行体外培养,解剖镜下观察并统计其生发泡破裂(GVBD)率和第一极体(PB1)排出率;免疫荧光染色、激光扫描共焦显微镜分析系统观察各组卵母细胞第1次减数分裂过程中纺锤体形成及染色体排列状况。结果 TCS浓度≤15mg/L时,卵母细胞GVBD率、PB1排出率及到达第1次减数分裂中期(MⅠ)的卵母细胞百分率无明显变化;TCS浓度为30mg/L和60mg/L时,到达MⅠ期的卵母细胞百分率降低(P<0.05),纺锤体形成和染色体排列出现异常,PB1排出率低于对照组(P<0.05);TCS浓度达到120mg/L时,MⅠ期卵母细胞的百分率显著下降(P<0.01),GVBD率下降(P<0.05),浓度为240mg/L时明显下降(P<0.01)。结论 TCS干扰第1次减数分裂重启及纺锤体形成和染色体排列,阻滞小鼠卵母细胞第1次减数分裂,且这种阻滞作用呈浓度依赖性。
Objective To study the effects of trichosanthin (TCS) on the first meiosis of mouse oocytes. Methodically matured female Kunming mice, 48h after injection of pregnant mare serum gonadotropin 10IU, obtained from the ovary immature oocytes a total of 480, containing different concentrations of TCS in culture medium, cultured in vitro groups were observed under the microscope (GVBD) rate and the first polar body (PB1) excretion rate. The immunofluorescence staining and laser scanning confocal microscopy analysis system were used to observe the formation of spindles during the first meiosis of oocytes in each group and Chromosome arrangement. Results When the concentration of TCS was less than 15mg / L, the oocytes showed no significant changes in GVBD rate, PB1 release rate and the percentage of oocytes reaching the first meiotic metaphase (MⅠ). When TCS concentrations were 30mg / L and 60mg / L (P <0.05), spindle formation and chromosomal arrangement were abnormal, and PB1 release rate was lower than that of the control group (P <0.05). When TCS concentration was 120 mg / L, the expression of MⅠ oocyte The percentage of cells decreased significantly (P <0.01), and the GVBD rate decreased (P <0.05). The concentration of 240 mg / L decreased significantly (P <0.01). Conclusion TCS interfered with the first meiotic resumption and spindle formation and chromosome arrangement, blocking the first meiosis of mouse oocytes, and the blocking effect was concentration-dependent.