摘要
[目的]观察育麟方对老龄导致卵巢储备功能下降(diminished ovarian reserve,DOR)小鼠卵母细胞数量与体外受精率的影响。[方法]将70只6～7月龄的C57BL/6J雌性小鼠随机分为育麟方高剂量组、育麟方中剂量组、育麟方低剂量组、脱氢表雄酮(dehydroepiandrosterone,DHEA)组和老龄对照组;另外14只C57BL/6J性成熟雌性小鼠作为青年对照组。育麟方高、中、低剂量组分别用25.0g/kg·d、12.0g/kg·d、6.0g/kg·d育麟方浓缩液灌胃,DHEA组用10.2mg/kg·d的DHEA灌胃,青年对照组和老龄对照组每日均采用蒸馏水0.2mL/10g灌胃,共用药40d。用药第38天开始腹腔序贯注射孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG)10U、人绒毛膜促性腺激素(human choionic gonadotophin,HCG)10U行控制性超促排卵,每组选择7只小鼠收集并比较卵母细胞数量;余下7只与C57BL/6J青年雄性小鼠行体外受精(in vitro fertilization,IVF),观察体外受精率;收集各组小鼠卵巢组织,并采用Western blot检测β-catenin、Wnt4蛋白表达。[结果]与青年对照组小鼠比较,老龄对照组小鼠卵巢指数、卵母细胞数量和成熟卵率显著下降,差异具有统计学意义(P<0.01),受精率显著下降(P<0.01)。与老龄对照组比较,育麟方高剂量组、DHEA组小鼠卵巢指数明显升高(P<0.01),且育麟方高剂量组高于DEHA组(P<0.01);与老龄对照组比较,育麟方高剂量组、DHEA组卵母细胞数显著上升(P<0.01),且育麟方高剂量组高于DHEA组(P<0.05);育麟方高剂量组小鼠卵母细胞成熟卵率高于老龄对照组(P<0.05)。[结论]高剂量育麟方可以改善老龄所致的卵巢指数下降、增加卵母细胞数量和成熟卵率,从而提高体外受精率。
[Objective]To observe the effect of Yulin formula on the number of oocytes and in vitro fertilization(IVF)rate in aged mice with diminished ovarian reserve(DOR).[Methods]Seventy female C57 BL/6 J mice aged from 6 to 7 months were randomly divided into five groups:Yulin formula high dose group,Yulin formula middle dose group,Yulin formula low dose group,dehydroepiandrosterone(DHEA)group and aged control group;and other 14 mature female C57 BL/6 J mice were used as young control group.Yulin formula high,middle and low dose groups were given Yulin formula 25.0 g/kg·d,12.0 g/kg·d and 6.0 g/kg·d,respectively;while the DHEA group was given DHEA 10.2 mg/kg·d,young control group and aged control group were given0.2 mL/10 g distilled water daily for 40 days.Sequential intraperitoneal injection of pregnant mare serum gonadotrophin(PMSG)and human chorionic gonadotrophin(HCG)were started on the 38 th day.Oocytes were collected and compared in 7 mice in each group;7 young male mice were fertilized in vitro to observe the fertilization rate in vitro;ovarian tissues were collected from each group and ovarian tissue was collected.The expression ofβ-catenin and Wnt4 was detected by Western blot.[Results]Compared with young control group,the ovarian index,the number of oocytes and the metaphaseⅡ(MⅡ)rate of aged control group decreased significantly(P<0.01),and the fertilization rate decreased significantly(P<0.01).Compared with aged control group,the ovarian index of Yulin formula high dose group and DHEA group increased significantly(P<0.01),and it was higher in Yulin formula high dose group than DEHA group(P<0.01);the number of oocyte in Yulin formula high dose group and DHEA group increased significantly(P<0.01),and it was higher in Yulin formula high dose group than DHEA group(P<0.05);the MⅡrate of Yulin formula high dose group was higher than aged control group(P<0.05).[Conclusion]High dose Yulin formula can increase the ovarian index,the number of oocyte and MⅡrate,raise the fertilization rate of IVF.
作者
马景
张驰
章勤
何嘉琳
刘晓昱
缪晨韵
MA Jing;ZHANG Chi;ZHANG Qin(Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310007,China;Zhejiang Chinese Medical University;Wenzhou Hospital of Traditional Chinese Medicine)
出处
《浙江中医药大学学报》
CAS
2019年第3期205-211,共7页
Journal of Zhejiang Chinese Medical University
基金
杭州市科技发展计划项目(20150733Q58)~~
