CLINICAL TRIAL: METFORMIN ON CHANGES IN AMPKINASE ACTIVITY IN PERIPHERAL BLOOD

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 目的：腺苷单磷酸活化蛋白激酶（AMPK）是细胞能量状态的标志物，通过催化亚基内Thr 172的磷酸化而活化。 已知二甲双胍诱导AMPK活性。AMPK激活发生在外周血单个核细胞体外 (PBM)，但没有在人PBM中的体内研究。 本研究将确定人PBM是否可用作测量二甲双胍诱导的AMPK活性变化的非侵入性方法。 由于胰岛素抵抗（IR）与AMPK活性降低相关，因此本研究还将确定瘦型胰岛素敏感（IS）、肥胖IS和肥胖IR受试者之间基线时和二甲双胍应答时人PBM中AMPK活性是否存在差异。 参与者：18- 55岁非糖尿病瘦和肥胖男性和女性 程序（方法）：在二甲双胍治疗前、治疗后2天（48小时）和治疗后4天（96小时）采集血样。 分离单核细胞。对裂解物进行电泳以分离蛋白质，然后用总AMPK和磷酸化AMPK <$Thr 172的抗体进行免疫印迹。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Purpose: Adenosine monophosphate-activated protein kinase (AMPK), a marker of cellular energy status, is activated by phosphorylation of Thr 172 within the catalytic ¿ subunit. Metformin is known to induce AMPK activity. AMPK activation occurs in vitro in peripheral blood mononuclear cells (PBM), but there are no in vivo studies in human PBM. This study will determine if human PBM can be used as a noninvasive method of measuring changes in AMPK activity induced by metformin. As insulin resistance (IR) is associated with decreased AMPK activity, this study will also determine if there is a difference in AMPK activity in human PBM at baseline and in response to metformin between lean insulin sensitive (IS), obese IS, and obese IR subjects. Participants: 18-55yo nondiabetic lean and obese men and women Procedures (methods): Obtain blood samples before, after 2 days (48hrs), and after 4 days (96hrs) of metformin therapy. Isolate the mononuclear cells. Perform electrophoresis of the lysate to separate proteins, then immunoblot with antibody to both total AMPK and phospho-AMPK ¿ Thr 172.