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利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：腺苷单磷酸激活的蛋白激酶(AMPK)是细胞能量状态的标志，通过催化亚基内苏氨酸172的磷酸化而被激活。已知二甲双胍可诱导AMPK活性。AMPK在体外可在外周血单核细胞中激活 (PBM)，但目前还没有关于人PBM的体内研究。这项研究将确定人的PBM是否可以作为一种非侵入性方法来测量二甲双胍诱导的AMPK活性的变化。由于胰岛素抵抗(IR)与AMPK活性降低有关，这项研究还将确定瘦胰岛素敏感(IS)、肥胖IS和肥胖IR受试者之间，人外周血中AMPK活性在基线和对二甲双胍的反应中是否存在差异。 参与者：18-55岁的非糖尿病瘦身和肥胖男性和女性 程序(方法)：在二甲双胍治疗前、治疗后2天(48小时)和治疗4天(96小时)后采集血样。分离单个核细胞。对裂解产物进行电泳法分离蛋白质，然后用总AMPK和磷酸化AMPK的抗体进行免疫印迹172。

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.