| 摘要: | 多囊性卵巢症候群 (polycystic ovary syndrome, PCOS)為一種由許多外來因素所造成的女性生殖系統合併內分泌疾病。先前文獻指出PCOS患者月經週期不規律及高雄性激素血症與高血糖、血脂異常和氧化壓力呈正相關，目前治療此疾病方式多為藥物或手術，但可能造成許多副作用。許多研究發現天然物中酚酸類化合物透過抗氧化功能可改善PCOS相關症狀，而咖啡酸 (caffeic acid, CA)為存在於蔬果中之酚酸類化合物，具有抗氧化及調節血糖之功效，但目前無相關文獻針對咖啡酸對PCOS改善效應之研究。因此，本研究欲了解咖啡酸對PCOS症狀之改善效果及其相關分子機制探討。體外試驗使用人類卵巢顆粒細胞株 (ovarian granulosa cells line, KGN)，透過過氧化氫 (hydrogen peroxide, H2O2)誘導細胞凋亡。以3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)assay測定細胞存活率，2',7'-dichlorodihydrofluorescein diacetate測定胞內活性氧物種 (reactive oxygen species, ROS)螢光強度。使用脫氫表雄酮 (dehydroepiandrosterone, DHEA)誘導雌性SD大鼠PCOS動物模式，誘導成功後分別介入咖啡酸及metformin 6週，以Vaginal smear觀察動情週期變化，分析血清賀爾蒙濃度、血糖、血脂及組織氧化壓力，以Western blot測定固醇類激素合成酵素及凋亡相關蛋白表現量。結果顯示，咖啡酸可顯著抑制ROS生成並保護KGN免受氧化壓力誘導細胞凋亡。透過體內試驗發現，咖啡酸可改善PCOS大鼠動情週期紊亂、空腹血糖、肝功能及血脂，降低血清過多雄性激素，並調節固醇類激素合成酵素及改善凋亡相關蛋白表現量，顯示咖啡酸具有改善PCOS併發症之潛力。
Polycystic ovary syndrome (PCOS), an endocrine disease, mainly occurs in reproductive age women, causing reproductive dysfunction and metabolic abnormalities. Previous literature had pointed out that irregular menstrual cycle and hyperandrogenism in PCOS patients were related to hyperglycemia, dyslipidemia, and oxidative stress. However, clinical treatments to PCOS are using drug or surgery, but these might cause side effects. Many studies had found that natural phenolic acids had antioxidant functions and could improve PCOS-related symptoms, while caffeic acid, a phenolic acid, existed in fruits and vegetables, having antioxidant and blood sugar regulation function. But there was no relevant research in the improvement effect of caffeic acid on PCOS. The aim of this study was to investigate whether caffeic acid could improve the systems of PCOS and its related molecular mechanism. In vitro study, using human ovarian granulosa cells line (KGN cells) to induce apoptosis by hydrogen peroxide (H2O2). Cell viability was determined by MTT assay, and intracellular reactive oxygen species (ROS) fluorescence intensity was determined by 2',7'-dichlorodihydrofluorescein diacetate. In vivo study, using dehydroepiandrosterone to induce PCOS animal model in female SD rats and caffeic acid and metformin were intervened for 6 weeks. Estrous cycles were observed by vaginal smear, sex hormone was measured by ELISA kits and tissue oxidative stress was measured by TBARS assay. Steroidogenesis enzyme and apoptosis-related protein expression were measured by western blot. The result showed that caffeic acid could significantly inhibit ROS generation, protect KGN from oxidative stress and H2O2 induced cell-apoptosis-related protein expression. In vivo experiments, caffeic acid could improve irregular estrous cycle, fasting blood glucose, liver function, and lipid profile in DHEA-induced PCOS rats. Besides, it also reduced hyperandrogenism and improved steroidogenesis enzyme and apoptosis-related protein expression. Therefore, our findings suggest that caffeic acid have the potential effect to improve PCOS complications. |
