Inflammation: Effect on Insulin Resistance in PCOS

描述（由申请人提供）：多囊卵巢综合征（PCOS）是一种病因不明的内分泌疾病，影响多达8%的女性人口，是女性不孕症的最常见原因。PCOS的慢性无排卵和高雄激素血症特征是由胰岛素抵抗和大多数受影响个体中产生的高胰岛素血症引起的。胰岛素抵抗使那些患有2型糖尿病的人更容易患上2型糖尿病，并且是心血管疾病早发的已知危险因素。肿瘤坏死因子α（TNF-α）是一种由脂肪组织和活化的单核细胞（MNC）产生的炎性细胞因子，其被认为是PCOS中胰岛素抵抗的介导者。拟议的研究将涉及对40名育龄妇女（18-40岁）的前瞻性研究。其中20名妇女将患有PCOS（10名正常体重和10名肥胖），20名将是体重匹配的排卵对照组。中心假设是生理性碳水化合物挑战触发PCOS妇女MNC过度释放TNF-α。具体目标是：1）检查PCOS女性MNC响应于葡萄糖刺激的TNF-α释放; 2）检查PCOS女性MNC响应于葡萄糖刺激的炎症途径。该方法涉及在标准口服葡萄糖激发试验期间以及在正常血糖和高血糖条件下，使用从空腹血液样品分离的培养的MNC，在体外对PCOS妇女的MNC培养上清液在体内葡萄糖暴露之前和之后的TNF-α释放进行定量。还将在体内葡萄糖激发之前和之后通过测量[活性氧簇] ROS产生、炎症途径的蛋白质标志物和活化的核因子κ B（NF κ B）的表达来评价PCOS女性中MNC的炎症反应。我们预期，本研究中使用的方法将证明在葡萄糖刺激后MNC衍生的TNF-α释放过量和明显的炎症反应。这些结果将是重要的，因为它们将确定MNC作为除脂肪组织外PCOS中过量TNF-α的额外来源，以及负责从MNC过度释放TNF-α的炎症机制。这将导致PCOS治疗的重要进展，旨在通过减少炎症来改善胰岛素抵抗。它也将为未来的研究提供优先权，以确定负责MNC的敏感性的分子机制，以餐后高血糖的PCOS妇女，这些人在一个促炎症状态。

DESCRIPTION (provided by applicant): Polycystic Ovary Syndrome (PCOS) is an endocrinopathy of unknown etiology that affects as many as 8% of the female population and is the most common cause of female infertility. The chronic anovulation and hyperandrogenism characteristic of PCOS is promulgated by insulin resistance and the resultant hyperinsulinemia in the majority of affected individuals. The insulin resistance predisposes those with the disorder to type 2 diabetes and is a known risk factor for early onset of cardiovascular disease. Tumor necrosis factor alpha (TNF-alpha), an inflammatory cytokine produced by adipose tissue and activated mononuclear cells (MNC) has been implicated as a mediator of insulin resistance in PCOS. The proposed research will involve the perspective study of 40 reproductive age women (18-40 years). Twenty of the women will have PCOS (10 normal weight and 10 obese) and 20 will be weight-matched ovulatory controls. The central hypothesis is that a physiologic carbohydrate challenge triggers excessive TNF-alpha release from MNC of women with PCOS. The specific aims are: 1) to examine TNF-alpha release from MNC of women with PCOS in response to glucose stimulation; 2) to examine the inflammation pathway in MNC of women with PCOS in response to glucose stimulation. The approach involves the quantification of TNF-alpha release from MNC culture supernatants of women with PCOS before and after in vivo glucose exposure during a standard oral glucose challenge test and under euglycemic and hyperglycemic conditions in vitro using cultured MNC isolated from fasting blood samples. The inflammatory response of MNC in women with PCOS will also be evaluated before and after in vivo glucose challenge by measuring [reactive oxygen species] ROS generation, expression of protein markers of the inflammation pathway and activated nuclear factor kappa B (NFkB). It is our expectation that the approach used in this study will demonstrate excessive MNC-derived TNF-alpha release and a pronounced inflammatory response following glucose stimulation. These results will be significant in that they will identify the MNC as an additional source of excess TNF-alpha in PCOS aside from adipose tissue, and the inflammatory mechanism responsible for excessive TNF-alpha release from MNC. This will lead to important advances in the therapy of PCOS designed to ameliorate insulin resistance by reducing inflammation. It will also provide precedence for future studies to determine the molecular mechanism responsible for the sensitivity of MNC to postprandial hyperglycemia in women with PCOS that places these individuals in a proinflammatory state.