Protective Effects of Puerarin on Premature Ovarian Failure via Regulation of Wnt/β-catenin Signaling Pathway and Oxidative Stress
Protective Effects of Puerarin on Premature Ovarian Failure via Regulation of Wnt/β-catenin Signaling Pathway and Oxidative Stress
复制标题
DOI：
10.1007/s43032-020-00325-0
复制
发表时间：
2020-09-29
影响因子：
2.9
通讯作者：
Huang, Jian
中科院分区：
文献类型：
--
作者：
Chen, Cheng;Li, Song;Huang, Jian
本研究旨在探讨葛根素（PUE）通过Wnt/ β -catenin信号通路和氧化应激对卵巢早衰（POF）模型的保护作用。两个月大的雌性小鼠被随机分为四组。1组作为对照组，其余3组分别注射环磷酰胺和丁硫凡建立POF模型。POF治疗组分别以100、200 mg/kg PUE灌胃28 d。固定卵巢，测定不同时期卵泡数量，收集卵巢表面上皮细胞（OSE）。采用免疫组化、RT-QPCR和western blotting检测Oct4和Mvh表达、Wnt/ β -catenin信号通路活性、氧化应激因子SOD2和Nrf2以及凋亡相关蛋白Bcl-2和Bax。我们发现，在POF组中，卵泡数量、Oct4和Mvh的表达以及Wnt/ β -连环蛋白信号活性均降低（p < 0.05或p < 0.001）。PUE治疗后，卵泡数和原始卵泡比增加（p < 0.01），闭锁比降低（p < 0.01）。与POF组相比，Oct4、Mvh、Wnt1、β -catenin、cyclin D1、SOD2、Nrf2的表达水平均有明显恢复（p < 0.01,p < 0.05，或p < 0.001）。与对照组相比，POF模型的Bcl-2/Bax比值降低了约60% (p < 0.001)， PUE治疗后Bcl-2/Bax比值提高了约50% （p < 0.001）。综上所述，PUE可能通过Wnt/ β -catenin信号通路提高女性生殖干细胞（FGSCs）的存活率，并对POF发挥保护作用，同时缓解氧化应激。进一步的研究应该集中在体外培养卵母细胞和FGSCs的PUE环境中，使用Wnt信号通路的抑制剂或激动剂。
This study was designed to investigate the protective effects of puerarin (PUE), which work via the Wnt/beta-catenin signaling pathway, and oxidative stress in the premature ovarian failure (POF) model. Two-month-old female mice were randomly divided into four groups. One group was used as the control, and the other three groups were injected with cyclophosphamide and busulfan to create POF models. Two POF treatment groups were gavaged with 100 or 200 mg/kg PUE for 28 days. Next, the ovaries were fixed, and the numbers of different stage follicles were measured, and the ovarian surface epithelium (OSE) was collected. Oct4 and Mvh expression, Wnt/beta-catenin signaling pathway activity, the oxidative stress factors SOD2 and Nrf2, and the apoptosis-related proteins Bcl-2 and Bax were detected by IHC, RT-QPCR, and western blotting. We found that the number of follicles, Oct4 and Mvh expression, and Wnt/beta-catenin-signaling activity were reduced in the POF groups (p < 0.05 orp < 0.001). After PUE treatment, the follicle number and the primordial follicle ratio increased (p < 0.01), while the atresia ratio decreased (p < 0.01). In addition, the expression levels of Oct4, Mvh, Wnt1, beta-catenin, cyclin D1, SOD2, and Nrf2 showed obvious recovery compared with levels in the POF group (p < 0.01,p < 0.05, orp < 0.001). The Bcl-2/Bax ratio in the POF model had reduced by about 60% compared with the control group (p < 0.001) and improved by about 50% after PUE treatment (p < 0.001). In conclusion, PUE may improve the survival of female reproductive stem cells (FGSCs) and play a protective role against POF via a mechanism involving the Wnt/beta-catenin signaling pathway, as well as relieving oxidative stress. Further investigations should focus on the culture of oocytes and FGSCs in vitro in a PUE environment with inhibitors or agonists of the Wnt signaling pathway.