| 马景,张驰,章勤,等.育麟方改善老龄雌性小鼠超促排卵作用的机制研究[J].浙江中医药大学学报,2019,43(3):205-211. |
| 育麟方改善老龄雌性小鼠超促排卵作用的机制研究 |
| Study on the Mechanism of Yulin Formula on Improving Controlled Ovarian Hyperstimulation in Aged Female Mice |
| DOI：10.16466/j.issn1005-5509.2019.03.001 |
| 中文关键词: 育麟方 老龄 卵巢储备功能低下 小鼠 体外受精 卵母细胞 |
| 英文关键词: Yulin formula aged diminished ovarian reserve mice in vitro fertilization oocyte |
| 基金项目:杭州市科技发展计划项目(20150733Q58) |
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| 中文摘要: |
| [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。 |
| 英文摘要: |
| [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 C57BL/6J 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 C57BL/6J mice were used as young control group. Yulin formula high, middle and low dose groups were given Yulin formula 25.0g/kg·d, 12.0g/kg·d and 6.0g/kg·d, respectively; while the DHEA group was given DHEA 10.2mg/kg·d, young control group and aged control group were given 0.2mL/10g distilled water daily for 40 days. Sequential intraperitoneal injection of pregnant mare serum gonadotrophin(PMSG) and human chorionic gonadotrophin(HCG) were started on the 38th 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 of β-catenin and Wnt4 was detected by Western blot. [Results] Compared with young control group, the ovarian index, the number of oocytes and the metaphase Ⅱ(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 MⅡ rate 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 MⅡ rate, raise the fertilization rate of IVF. |
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