温肾养血方对高龄雌鼠卵泡发育的作用及其对PI3K-AKT-FOXO3a通路影响的研究

doi:10.3969/j.issn.1006-7795.2019.03.005

首都医科大学学报 ›› 2019, Vol. 40 ›› Issue (3): 344-351.doi: 10.3969/j.issn.1006-7795.2019.03.005

温肾养血方对高龄雌鼠卵泡发育的作用及其对PI3K-AKT-FOXO3a通路影响的研究

辛明蔚¹, 张莹¹, 何军琴¹, 杨维¹, 武颖¹, 尹晓丹¹, 贾朝霞²

1. 首都医科大学附属北京妇产医院中医科, 北京 100026;
2. 首都医科大学附属北京妇产医院群体保健科, 北京 100026

收稿日期:2019-03-15 出版日期:2019-05-21 发布日期:2019-06-13
通讯作者: 张莹 E-mail:zhangying118800@163.com
基金资助:
国家自然科学基金（81503607），北京市医院管理局临床医学发展专项经费资助（ZYLX201510），北京市医院管理局登峰人才培养计划（DFL20151301）。

Effect of Wenshen Yangxue Decoction on follicular development in aged female rats and its impact on PI3K-AKT-FOXO3a pathway

Xin Mingwei¹, Zhang Ying¹, He Junqin¹, Yang Wei¹, Wu Ying¹, Yin Xiaodan¹, Jia Zhaoxia²

1. Department of Traditional Chinese Medicine, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, China;
2. Information and Statistics Department, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, China

Received:2019-03-15 Online:2019-05-21 Published:2019-06-13
Supported by:
This study was supported by National Natural Science Foundation of China (81503607),Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support (ZYLX201510),Beijing Municipal Administration of Hospitals'Ascent Plan (DFL20151301).

摘要/Abstract

摘要： 目的探讨温肾养血方促进高龄雌鼠卵泡发育的作用及机制。方法采用数字表法将8月龄ICR小鼠随机分为A：空白组、B：控制性超排卵组（controlled ovarian hyperstimulation，COH）组、C：温肾养血方低剂量组、D：温肾养血方中剂量组、E：温肾养血方高剂量组，连续每日定时（下午6时）灌胃给药10 d，第11天对B、C、D、E组雌鼠同时腹腔注射孕马血清促性腺激素（pregnant mare serum gonadotropin，PMSG）10 IU/只，A、B组雌鼠同时注射等量0.9%（质量分数）氯化钠注射液，48 h后，腹腔注射人绒毛膜促性腺激素（human chorionic gonadotropin，HCG）10 IU/只，14 h后，剖腹取出小鼠卵巢及输卵管，观察用药后各组小鼠取卵数及优质卵泡数，qRT-PCR、Western blotting法和免疫组织化学法检测PI3K、P-PI3K、AKT、P-AKT、FOXO3a、P-FOXO3a的表达情况。每组记录4只雌鼠与雄鼠交配后的生育数量。结果温肾养血方能够增加小鼠排卵数、优质卵泡数和生育数；与空白组比较，温肾养血方中剂量组小鼠卵巢PI3K mRNA表达下降，温肾养血方高、低剂量组表达升高，差异有统计学意义（P<0.05）；低剂量组AKT mRNA的表达升高，差异有统计学意义（P<0.05）；低剂量、中剂量和高剂量组的FOXO3a mRNA表达均升高，差异有统计学意义（P<0.05），随着中药浓度升高，FOXO3a mRNA表达的含量逐渐下降。结论温肾养血方可促进高龄ICR小鼠卵泡发育，增加排卵数、优质卵泡数及生育数量，其机制可能与调节PI3K，AKT和FOXO3a的表达有关。温肾养血方低剂量和中剂量效果最好，高剂量次之。

关键词: 温肾养血方, PI3K-AKT-FOXO3a通路, 控制性超排卵

Abstract: Objective To explore the effect and mechanism of Wenshen Yangxue Decoction on promoting follicular development in aged female rats. Methods Eight-month-old ICR mice were randomly divided into 5 groups:A:blank group, B:controlled ovarian hyperstimulation (COH) group, C:low dose group of Wenshen Yangxue Decoction, D:middle dose group of Wenshen Yangxue Decoction, and E:high dose group of Wenshen Yangxue Decoction, and given intragastric administration for 10 days continuously at regular intervals (6:00 pm). On the 11th day, female mice in the groups B, C, D and E were intraperitoneally injected with pregnant mare serum gonadotropin (PMSG) 10 IU/mouse. Female mice in the groups A and B were intraperitoneally injected with human chorionic gonadotropin (HCG) 10 IU/mouse 48 hours later. At 14 hours later, ovaries and oviducts of mice were removed by laparotomy. After treatment, number of follicles and number of high quality follicles were examined, and the expressions of PI3K, P-PI3K, AKT, P-AKT, FOXO3a and P-FOXO3a were measured with qRT-PCR, Western blotting, and immunohistochemistry. The number of births after mating of 4 females and males was recorded for each group.Results Wenshen Yangxue Decoction could increase the number of ovulation, the number of high quality follicles and fertility of mice. Compared with blank group, the expression of PI3K mRNA in the ovary of mice in Wenshen Yangxue Decoction middle-dose group decreased, while the expression in Wenshen Yangxue Decoction high-dose and low-dose groups increased, with the difference statistically significant (P<0.05). The expression of AKT mRNA in low-dose group increased, with the difference statistically significant (P<0.05). The expression of FOXO3a mRNA in low-dose, medium-dose and high-dose groups increased, with the difference statistically significant (P<0.05), as the concentration of the TCM increased, the content of FOXO3a mRNA expression gradually decreased. Conclusion Wenshen Yangxue Decoction can promote follicular development, increase the number of ovulation, quality follicle, and embryo number in aged ICR mice by regulating the expression of PI3K, AKT and FOXO3a. Low and medium doses were found to yield the most effective results, followed by high doses.

Key words: Wenshen Yangxue Decoction, PI3K-AKT-FOXO3a pathway, controlled ovarian hyperstimulation

中图分类号:

R984

辛明蔚, 张莹, 何军琴, 杨维, 武颖, 尹晓丹, 贾朝霞. 温肾养血方对高龄雌鼠卵泡发育的作用及其对PI3K-AKT-FOXO3a通路影响的研究[J]. 首都医科大学学报, 2019, 40(3): 344-351.

Xin Mingwei, Zhang Ying, He Junqin, Yang Wei, Wu Ying, Yin Xiaodan, Jia Zhaoxia. Effect of Wenshen Yangxue Decoction on follicular development in aged female rats and its impact on PI3K-AKT-FOXO3a pathway[J]. Journal of Capital Medical University, 2019, 40(3): 344-351.

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温肾养血方对高龄雌鼠卵泡发育的作用及其对PI3K-AKT-FOXO3a通路影响的研究

Effect of Wenshen Yangxue Decoction on follicular development in aged female rats and its impact on PI3K-AKT-FOXO3a pathway

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