芪苓温肾消囊颗粒调控亮氨酸代谢诱导mTOR/S6K1通路治疗PCOS胰岛素抵抗机制研究

Polycystic ovary syndrome (PCOS) is a common gynecological reproductive endocrine and metabolic disorder. Insulin resistance is an important pathological basis for the occurrence and development of PCOS, which is prevalent in PCOS women with various body mass indexes. Previous studies have found that leucine is the strongest predictor of insulin sensitivity reduction, which helps to monitor the development of insulin resistance. In this study, we established PCOS rat models with insulin resistance. We used Qiling wenshen xiaonang granule (Q) combined with leucine deficient diet or whole amino acid diet intervention. The endocrine, amino acid metabolism, and the expressions of mTOR, S6K1 and insulin signal transduction key proteins in tissues were detected by HPLC and western-blot, etc. We prepared S6K1 recombinant adenovirus and injected it into model rats to observe the effect of S6K1 gene silencing on insulin signal transduction. To elucidate the effect of leucine on insulin sensitivity in PCOS with insulin resistance. To explore whether this prescription can to induce mTOR/S6K1 pathway to increase PCOS insulin sensitivity by regulating the metabolism of bright amino acids, so as to provide molecular biological evidence for the treatment of PCOS insulin resistance by this prescription.

多囊卵巢综合征（PCOS）是妇科常见生殖内分泌代谢异常疾病，胰岛素抵抗是本病发生发展的重要病理基础，普遍存在于各体重指数的PCOS女性中。既往研究发现，亮氨酸可作为胰岛素敏感性降低最强代谢预测因子，有助于监测胰岛素抵抗的发展。本研究建立PCOS伴胰岛素抵抗大鼠模型，以芪苓温肾消囊颗粒配合亮氨酸缺乏饮食或全氨基酸饮食干预，运用高效液相色谱法、Western-Blot等检测方法，分析内分泌、氨基酸代谢，检测组织中mTOR、S6K1及胰岛素信号传导关键蛋白表达。制备S6K1重组腺病毒，注入模型大鼠观察S6K1基因沉默后对胰岛素信号传导的影响。阐明胰岛素抵抗条件下，亮氨酸对PCOS胰岛素抵抗的作用。探讨芪苓温肾消囊颗粒可通过调节亮氨酸代谢从而诱导mTOR/S6K1通路增加PCOS胰岛素敏感性，为本方治疗PCOS胰岛素抵抗提供分子生物学证据。

多囊卵巢综合征（PCOS）是女性常见的生殖内分泌代谢异常疾病。胰岛素抵抗是PCOS发生发展的重要病理基础，普遍存在于各体重指数的PCOS女性中。本研究通过大样本PCOS患者临床数据分析PCOS患者糖代谢特点，明确高雄激素对于PCOS糖代谢的影响。研究通过纳入PCOS代谢异常患者、PCOS女性及健康女性，检测临床及血清代谢组学结果，明确血清亮氨酸水平与PCOS发生胰岛素抵抗的相关性，及血清亮氨酸对PCOS代谢异常的诊断价值。采用PCOS胰岛素抵抗大鼠模型，以芪苓温肾消囊颗粒（QL颗粒）配合亮氨酸缺乏饮食干预。结果显示QL颗粒、限制亮氨酸摄入均可降低血清亮氨酸水平，减轻PCOS胰岛素抵抗模型大鼠高胰岛素血症、改善胰岛素抵抗，提高全身胰岛素敏感性。联合应用QL颗粒、限制亮氨酸摄入可显著降低血清总睾酮水平。通过检测PCOS胰岛素抵抗大鼠经肝脏门静脉注射胰岛素干预前后肝脏、腹部脂肪、比目鱼肌、卵巢组织中IR-β、AKT等蛋白表达，结果显示QL颗粒、限制亮氨酸摄入均可增加肝脏、脂肪、肌肉、卵巢组织外周组织胰岛素敏感性。PCOS持续睾酮暴露增加胰岛素引起的mTOR和S6K1磷酸化。S6K1腺病毒（Ad-S6K1）干预后，S6K1活性升高降低全身胰岛素敏感性，经门静脉注射胰岛素后肝脏、卵巢组织IR-β和p-AKT/AKT比值降低，表明Ad-S6K1抑制QL颗粒、限制亮氨酸摄入对全身和局部胰岛素敏感性的作用。本研究明确了PCOS胰岛素抵抗患者血清亮氨酸水平升高，限制亮氨酸摄入可改善PCOS胰岛素抵抗。QL颗粒通过调控亮氨基酸代谢影响AKT/mTOR/S6K1途径改善胰岛素敏感性，PCOS体内高雄激素和胰岛素抵抗协同作用，其作用超过了单独胰岛素抵抗。