Shanghai Geriatric Institute of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Department of Pathology, Yale University School of Medicine, New Haven, CT, USA.
Cell Prolif. 2021 Jan;54(1):e12954. doi: 10.1111/cpr.12954. Epub 2020 Nov 9.
To examine the role of high-fat and high-sugar (HFHS) diet-induced oxidative stress, which is a risk factor for various diseases, in premature ovarian failure (POF).
Ovarian granulosa cells (OGCs) were isolated from mice and cultured in medium supplemented with HFHS and poly (lactic-co-glycolic acid) (PLGA)-cross-linked miR-146b-5p nanoparticles (miR-146@PLGA). RNA and protein expression levels were examined using quantitative real-time polymerase chain reaction and Western blotting, respectively. HFHS diet-induced POF model mice were administered miR-146@PLGA.
The ovarian tissue of mice fed a HFHS diet exhibited the typical pathological characteristics of POF. HFHS supplementation induced oxidative stress injury in the mouse OGCs, activation of the Dab2ip/Ask1/p38-Mapk signalling pathway and phosphorylation of γH2A.X in vitro and in vivo. The results of the luciferase reporter assay revealed that miR-146 specifically downregulated p38-Mapk14 expression. Meanwhile, co-immunoprecipitation and Western blot analyses revealed that HFHS supplementation upregulated nuclear p38-Mapk14 expression and consequently enhanced γH2A.X (Ser139) phosphorylation. The HFHS diet-induced POF mouse model treated with miR-146@PLGA exhibited downregulated p38-Mapk14 expression in the OGCs, mitigated OGC ageing and alleviated the symptoms of POF.
This study demonstrated that HFHS supplementation activates the Dab2ip/Ask1/p38-Mapk signalling pathway and promotes γH2A.X phosphorylation by inhibiting the expression of endogenous miR-146b-5p, which results in OGC ageing and POF development.
探讨高脂肪高糖（HFHS）饮食诱导的氧化应激在卵巢早衰（POF）中的作用，氧化应激是多种疾病的危险因素。
从小鼠中分离卵巢颗粒细胞（OGC），并在补充 HFHS 和聚乳酸-羟基乙酸共聚物（PLGA）交联 miR-146b-5p 纳米颗粒（miR-146@PLGA）的培养基中培养。分别采用实时定量聚合酶链反应和 Western blot 检测 RNA 和蛋白质表达水平。用 miR-146@PLGA 处理 HFHS 饮食诱导的 POF 模型小鼠。
喂食 HFHS 饮食的小鼠卵巢组织表现出典型的 POF 病理特征。HFHS 补充物在体外和体内诱导了小鼠 OGC 的氧化应激损伤，激活了 Dab2ip/Ask1/p38-Mapk 信号通路和 γH2A.X 的磷酸化。荧光素酶报告基因检测结果表明，miR-146 特异性地下调了 p38-Mapk14 的表达。同时，共免疫沉淀和 Western blot 分析表明，HFHS 补充物上调了核 p38-Mapk14 的表达，从而增强了 γH2A.X（Ser139）的磷酸化。用 miR-146@PLGA 处理 HFHS 饮食诱导的 POF 小鼠模型，OGC 中的 p38-Mapk14 表达下调，OGC 衰老减轻，POF 症状缓解。
本研究表明，HFHS 补充物通过抑制内源性 miR-146b-5p 的表达激活了 Dab2ip/Ask1/p38-Mapk 信号通路，并促进了 γH2A.X 的磷酸化，导致 OGC 衰老和 POF 的发生。
