Nam Eun-Young, Kim Sun Ah, Kim Heejung, Kim Su-Hyun, Han Jae-Hyun, Lee Ju-Hee, Kim Dong-Il
Department of Korean Gynecology, College of Korean Medicine, Dongguk University, Goyang 10326, Republic of Korea.
College of Korean Medicine, Dongguk University, Goyang 10326, Republic of Korea.
J Ethnopharmacol. 2016 Dec 24;194:733-739. doi: 10.1016/j.jep.2016.10.048. Epub 2016 Oct 18.
Evodiae Fructus (EF) is the dried, unripe fruit of Evodia rutaecarpa Benth., and one of the main components of traditional herbal prescriptions issued for the treatment of sterility caused by irregular menstruation in Korea. However, scientific evidence regarding the efficacy and action mechanism of EF is lacking.
In this study, the authors established an in vitro screening tool to identify promising new drug candidates in herbal medicines for the prevention and treatment of premature ovarian failure. The protective effects of EF extracts against 4-vinylcyclohexene diepoxide (VCD)-induced ovotoxicity were investigated and the molecular mechanism responsible was sought.
EF extract was prepared by boiling EF in water and its quality was confirmed by high performance liquid chromatography. CHO-K1 (Chinese hamster ovary cells) and COV434 (human ovarian granulosa cells) cells were plated, pretreated with EF extract for 2h and then treated with 1.5mM or 0.5mM VCD for 24h, respectively. Cell viabilities were measured using an MTT assay, and protein levels were determined by western blotting.
VCD significantly suppressed the viability of both CHO-K1 and COV434 cells in a dose-dependent manner and induced the apoptosis of CHO-K1 cells at 1.5mM. EF extract dose-dependently blocked the ovotoxicity induced by treatment with VCD. Furthermore, EF extract significantly activated Akt and downstream effectors such as mTOR and GSK-3β in CHO-K1 cells. The ability of EF extract to prevent cytotoxicity by VCD was antagonized by pretreatment of LY294002, a PI3K/Akt inhibitor.
EF has the ability to protect ovary cells against VCD-induced ovotoxicity, probably via Akt activation. These results suggest that the beneficial effects of EF might be useful for preventing premature ovarian failure or unexplained infertility caused by environmental factors.
吴茱萸果实（EF）是吴茱萸干燥的未成熟果实，是韩国用于治疗月经不调所致不孕症的传统草药方剂的主要成分之一。然而，关于EF的功效和作用机制缺乏科学证据。
在本研究中，作者建立了一种体外筛选工具，以鉴定用于预防和治疗卵巢早衰的草药中有前景的新药候选物。研究了EF提取物对4-乙烯基环己烯二环氧化物（VCD）诱导的卵巢毒性的保护作用，并探寻其分子机制。
通过将EF水煮制备EF提取物，并通过高效液相色谱法确认其质量。接种中国仓鼠卵巢细胞（CHO-K1）和人卵巢颗粒细胞（COV43），分别用EF提取物预处理2小时，然后分别用1.5mM或0.5mM VCD处理24小时。使用MTT法测量细胞活力，并通过蛋白质印迹法测定蛋白质水平。
VCD以剂量依赖性方式显著抑制CHO-K1和COV434细胞的活力，并在1.5mM时诱导CHO-K1细胞凋亡。EF提取物剂量依赖性地阻断VCD处理诱导的卵巢毒性。此外，EF提取物显著激活CHO-K1细胞中的Akt及其下游效应分子，如mTOR和GSK-3β。PI3K/Akt抑制剂LY294002预处理可拮抗EF提取物预防VCD细胞毒性的能力。
EF具有保护卵巢细胞免受VCD诱导的卵巢毒性的能力，可能是通过激活Akt实现的。这些结果表明，EF的有益作用可能有助于预防由环境因素引起的卵巢早衰或不明原因的不孕症。
