左归丸对化疗源性卵巢早衰小鼠卵母细胞影响的实验研究

Experimental study on the effect of Zuogui Pill on the oocytes of chemotherapy-induced premature ovarian failure mice

目的研究左归丸对化疗源性卵巢早衰(premature ovarian failure,POF)小鼠卵母细胞以及其体外成熟培养(in vitro maturation,IVM)和体外受精实验(In vitro fertilization,IVF)结局的影响。方法以环磷酰胺(cyclophosphamide,CTX)建立化疗源性POF小鼠动物模型。提取各组生发泡(germinal vesicle,GV)期未成熟裸卵(denuded oocytes,DOs)及MII期卵母细胞分别进行IVM和IVF实验。观察左归丸对模型组小鼠DOs和MII期卵母细胞数量的影响、对IVM实验时生发泡破裂(germinal vesicle breakdown,GVBD)和第一极体(the first polar body,PB1)排出的影响及对IVF实验时2-细胞率和囊胚率的影响。使用激光扫描共聚焦显微镜观察各组IVM14h时MII期卵母细胞纺锤体和染色体形态。结果左归丸能增加化疗源性POF小鼠GV期DOs和MII期卵母细胞数量、显著提高模型组IVM实验的GVBD率和PB1排出率、显著提升模型组IVF实验的2-细胞率和囊胚率及纠正CTX引起的MII期卵母细胞纺锤体形成异常。结论左归丸可改善卵母细胞数量和质量,从而促进体外胚胎发育,对化疗源性POF小鼠起到一定治疗作用。

Objective To study the effect of Zuogui Pill on the oocytes and the outcome of IVM and IVF in mice with chemotherapy-induced premature ovarian failure(POF)of chemotherapy-induced premature ovarian failure(POF)mice.Methods The model of chemotherapy-induced POF was established by cyclophosphamide(CTX).Denuded oocytes(DOs)in the phase of germinal vesicle(GV)and MII oocytes were used for in vitro maturation(IVM)and in vitro fertilization(IVF)experiments respectively after medication.The effect of Zuogui Pill on the number of DOs in the phase of GV and MII phase,the germinal vesicle breakdown(GVBD)rate and the expulsion of first polar body(PB1)rate in IVF,the 2-cell rate and blastocyst rate in IVF were observed by data statistics.The spindle and chromosome morphology of MII oocytes were observed and evaluated by confocal laser scanning microscopy.Results Zuogui pill can increase the number of the number of DOs in the phase of GV and MII oocytes,increase the GVBD rate and PB1 excretion rate of IVM,increase the 2-cell rate and blastocyst rate of IVF,and correct the abnormal spindle formation of MII oocytes.Conclusion Zuogui pill can improve the quantity and quality of oocytes,promote the development of embryos in vitro,thus playing a therapeutic role in chemotherapy-induced POF mice.