逍遥丸对体外培养小鼠腔前卵泡卵母细胞骨形成蛋白6及其膜受体表达的影响  

Effects of Xiaoyao Pill on bone morphogenetic protein 6 and membrane receptors in oocytes of mouse preantral follicles cultured in vitro

作　　者：孙晓换[1,2] 杜惠兰 Sun Xiaohuan;Du Huilan(School of Traditional Chinese and Western Medicine,Hebei University of Chinese Medicine,Hebei 050091,China;Department of Obstetrics,Ningbo Medical Center Lihuili Eastern Hospital,Zhejiang 315000,China)

机构地区：[1]河北中医学院,河北050091 [2]宁波市医疗中心李惠利医院东部院区

出　　处：《北京中医药大学学报》2020年第3期218-224,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.81173294);河北省自然科学基金项目(No.H2015423050)。

摘　　要：目的观察逍遥丸对体外培养第6天小鼠腔前卵泡卵母细胞骨形成蛋白6(BMP6)及膜受体的影响,探讨其干预卵母细胞质量的可能机制。方法12只雌性SD大鼠随机分为逍遥丸高、中、低剂量组和对照组,分别灌服逍遥丸混悬液2.7、1.8、0.9 g/kg和等量蒸馏水,第4天股动脉采血,得到逍遥丸高、中、低剂量大鼠含药血清和正常大鼠血清。机械法显微分离12日龄雌性昆明小鼠腔前卵泡,分为逍遥丸高、中、低剂量组和对照组。逍遥丸组分别以高、中、低剂量逍遥丸大鼠含药血清孵育,对照组以正常大鼠血清孵育,血清浓度均为10%。细胞免疫荧光和实时荧光定量PCR分别检测体外培养第6天卵母细胞中BMP6、骨形成蛋白Ⅱ型受体(BMPRⅡ)、激活素受体样激酶(ALK)2、ALK6蛋白和mRNA表达。结果各组卵母细胞中均有BMP6、BMPRⅡ、ALK2、ALK6蛋白表达。与对照组比较,逍遥丸高、中、低剂量组BMP6、BMPRⅡ、ALK2、ALK6蛋白和mRNA表达均升高(P<0.05),以逍遥丸高剂量组最高(P<0.05)。结论逍遥丸可能通过上调卵母细胞中BMP6、BMPRⅡ、ALK2、ALK6表达,提高体外发育卵母细胞质量。Objective To observe the effects of Xiaoyao(Free Wanderer)Pill(XYP)on bone morphogenetic protein 6(BMP6)and membrane receptors in oocytes of mouse preantral follicles cultured in vitro and to explore the potential mechanisms of the intervention by XYP in the quality of oocytes.Methods Twelve female SD rats were randomly divided into four groups:high-,middle-and low-dose XYP groups and control group.XYP suspension(2.7,1.8 and 0.9 g/kg)were administered to the three XYP groups and the same amount of distilled water was given to the control group respectively for four days.By the end of the intervention,blood was drawn from the femoral artery to obtain rat serum containing XYP from the high-,middle-and low-dose XYP groups and normal serum from the control group.Primary preantral follicles of 60 female Kunming mice aged 12 days were micro-isolated mechanically.They were divided into high-,middle-,low-dose XYP groups and control group and incubated with 10%rat serum containing XYP from the high-,middle-and low-dose XYP rat groups and 10%normal serum from the control rat group respectively for six days.The protein and mRNA expressions of BMP6,BMPRⅡ,ALK2 and ALK6 in the oocytes were detected by cellular immunofluorescence and real-time reverse transcription PCR.Results Compared with the control group,the above protein and mRNA expressions in XYP groups were higher(P<0.05),and those in the high-dose XYP group were the highest(P<0.05).Conclusion XYP may improve the quality of oocytes cultured in vitro by increasing the expressions of BMP6,BMPRⅡ,ALK2,and ALK6 in the oocytes.

关 键 词：逍遥丸 骨形成蛋白6 膜受体 腔前卵泡 卵母细胞 小鼠 

分 类 号：R285.5[医药卫生—中药学]