多囊1号调控Kiss1/Kiss1R/Akt/FOXO1通路治疗多囊卵巢综合征的作用机制研究?? [ ]

Researchon on DNY Alleviate Polycystic Ovary Syndrome by Modulating the Kiss1/Kiss1R/Akt/FOXO1 Signaling Pathway

关键词:  多囊1号  多囊卵巢综合征  动物实验  性激素  

Key words:  DNY  polycystic ovary syndrome  animal experiment  sex hormone  

摘要:

目的 探讨多囊1号(DNY)治疗多囊卵巢综合征(PCOS)大鼠的潜在作用机制。方法 将雌性SD大鼠随机分为对照组、模型组、枸橼酸氯芬片组,多囊1号低、中、高剂量组,每组8只。采用连续灌胃来曲唑21天建立PCOS大鼠模型,并通过瑞氏染色法观察大鼠的动情周期,判断造模情况。造模成功后,对照组和模型组灌胃生理盐水,各给药组灌胃相应剂量的药物,持续14天。记录大鼠体重、卵巢质量,计算卵巢指数,HE染色观察卵巢组织的形态。ELISA法测定LH、FSH、T、AMH、E2、IL-6、MDA、SOD、Kiss1水平。实时荧光定量PCR检测卵巢组织FOXO1、CYP17A1、CYP19A1mRNA表达。免疫组化检测大鼠卵巢切片中的Kiss1 、P-Akt、FOXO1的表达情况。蛋白免疫印记法检测卵巢组织中的Kiss1 、Kiss1R、Akt、P-Akt、FOXO1蛋白的表达。结果 与模型组相比,DNY能降低大鼠的体重增长率,降低血清中LH、LH/FSH、T、AMH、IL-6、MDA、Kiss1水平,升高FSH、E2、SOD水平(P*＜0.05,P**＜0.01)。DNY给药后能改善卵巢组织病理形态,降低卵巢组织FOXO1、CYP17A1 mRNA表达,增加CYP19A1 mRNA表达(P**＜0.01),降低卵巢组织 Kiss1、FOXO1蛋白表达,增加Kiss1R、P-Akt/Ak蛋白的表达(P*＜0.05,P**＜0.01)。结论 多囊1号能够改善多囊卵巢综合征引起的性激素水异常,减轻卵巢组织的病理损伤,其作用机制可能与Kiss1/Kiss1R/AKT/FOXO1信号通路改善性激素分泌有关。

Abstract:

ABSTRACT: OBJECTIVE: To investigate the effect and mechanism of Polycystic 1（DNY）in improving polycystic ovary syndrome (PCOS) model rats . METHODS: Female SD rats were randomly divided into six groups: control group, model group, clomiphene citrate（CC） group, DNY low-dose group, DNY medium-dose group, and DNY high-dose group, with 8 rats in each group. A PCOS rat model was established by continuous intragastric administration of letrozole for 21 days, and the estrous cycle was observed using Wright's staining to evaluate the modeling status. After successful modeling, the normal control and model groups were given normal saline by intragastric gavage, while the treatment groups received corresponding doses of drugs for 14 days. Body weight and ovarian weight were recorded, and the ovarian index was calculated. Hematoxylin-Eosin (HE) staining was employed to observe ovarian histopathological changes. Serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), anti-Müllerian hormone (AMH), estradiol (E2), interleukin-6 (IL-6), malondialdehyde (MDA), superoxide dismutase (SOD), and Kiss1 were measured by using ELISA. Real-Time PCR was used to investigate the expression of FOXO1、CYP17A1、CYP19A1 mRNA in rats ovarian . The protein expression of Kiss1、P-Akt and FOXO1 were detected by Immunohistochemistry（IHC）.Western blotting（WB） was used to detect the protein expression of Kiss1, Kiss1R, Akt, phosphorylated Akt (P-Akt), and FOXO1 in ovarian tissue. RESULTS: Animal experimental results showed that compared with the model group, DNY reduced the body weight growth rate, decreased serum levels of LH, LH/FSH ratio, T, AMH, IL-6, MDA, and Kiss1, and increased serum FSH, E2, and SOD levels （P*＜0.05，P**＜0.01）. DNY improved ovarian pathological morphology. Compared with model group ,the expression level of FOXO1 and CYP17A1 mRNA were significantly decreased, the expression level of CYP19A1 mRNA was significantly increased（P**＜0.01）. Additionally, DNY downregulated the ovarian protein expression of Kiss1 and FOXO1, while upregulating the protein expression of Kiss1R and P-Akt/Akt ratio（P*＜0.05，P**＜0.01）. CONCLUSION: The traditional Chinese medicine compound DNY can improve sex hormone levels and alleviate ovarian pathological changes in PCOS rats. Its mechanism may be related to improving sex hormone secretion through the Kiss1/Kiss1R/AKT/FOXO1 signaling pathway.