【Jounal club】从 Nature medicine上一篇文章看蛋白质技术的运用
丁香园论坛
1022
最近看文献 深有感触啊
杂志及年卷期：Nat Med. 2002 Nov;8(11):1288-95. Epub 2002 Oct 7.
题目：Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase.
作者：Yamauchi T, 等.
Abstract：Adiponectin (Ad) is a hormone secreted by adipocytes that regulates energy homeostasis and glucose and lipid metabolism. However, the signaling pathways that mediate the metabolic effects of Ad remain poorly identified. Here we show that phosphorylation and activation of the 5'-AMP-activated protein kinase (AMPK) are stimulated with globular and full-length Ad in skeletal muscle and only with full-length Ad in the liver. In parallel with its activation of AMPK, Ad stimulates phosphorylation of acetyl coenzyme A carboxylase (ACC), fatty-acid oxidation, glucose uptake and lactate production in myocytes, phosphorylation of ACC and reduction of molecules involved in gluconeogenesis in the liver, and reduction of glucose levels in vivo. Blocking AMPK activation by dominant-negative mutant inhibits each of these effects, indicating that stimulation of glucose utilization and fatty-acid oxidation by Ad occurs through activation of AMPK. Our data may provide a novel paradigm that an adipocyte-derived antidiabetic hormone, Ad, activates AMPK, thereby directly regulating glucose metabolism and insulin sensitivity in vitro and in vivo.
中文翻译：
Adiponectin是一个调控能源体内平衡和葡萄糖和脂质新陈代谢的脂肪细胞因子。 然而, 其调控能源体内平衡和葡萄糖和脂质新陈代谢具体的作用机制不是很清楚。在这里我们证明了“磷酸化作用”和“AMPK”在球状和全长的Adiponectin在肌肉中以及全长Adiponectin在肝脏中的激活起着重要作用。 在激活 AMPK的同时, Adiponectin刺激了乙酰辅酶A羧化酶的磷酸化、脂肪酸氧化 、葡萄糖摄取和乳酸盐生成。乙酰辅酶A羧化酶的磷酸化和葡萄糖异生的产生与、又导致血糖的下降。阻断了AMPK的作用之后的结果显示了Adiponectin的促脂肪氧化 、葡萄糖利用增加作用均是ampk介导的。我们的实验在体内和体外均证实了这样一个新的范例。
个人感慨：这篇文章的内容我并不想多讲，是做一个蛋白的信号转导的。大家知道，日 本 人现在在国际顶尖的“CNS”杂志上发表文章很多，特别是蛋白做得很好，那么，大家可以看看他们做这篇文章用到了什么技术
我列举一下：
1、Generation of recombinant Ad. 蛋白的重组表达
2、Western-blot analysis
3、RNA preparation and northern-blot analysis.
4、Lipid and glucose metabolism. 属于最简单的生化指标（血糖血脂）测定
5、Studies with C2C12 myocytes and isolated hepatocytes
6、Adenovirus-mediated gene transfer in vivo and Ad tolerance test.
7、Measurement of AMP, ADP and ATP content. 放免法测定
各位战友看看，这些技术难不难？可能5、6项还算是有点难度的，而其他的工作是不是我们蛋白版里天天讨论的那些技术？日 本 人就是能善于利用普通技术，如Western，Northern，免疫组化，elisa这些，组合起来运用说明一个问题。选择的切入点、实验设计的巧妙性，这些真是我们要好好学习的。人家有这些技术为什么就能发Nature，而我们却还在为一两分的SCI苦苦拼搏。。值得深思啊
文章我一同附上了 大家看看 全是Western的图片。。。
杂志及年卷期：Nat Med. 2002 Nov;8(11):1288-95. Epub 2002 Oct 7.
题目：Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase.
作者：Yamauchi T, 等.
Abstract：Adiponectin (Ad) is a hormone secreted by adipocytes that regulates energy homeostasis and glucose and lipid metabolism. However, the signaling pathways that mediate the metabolic effects of Ad remain poorly identified. Here we show that phosphorylation and activation of the 5'-AMP-activated protein kinase (AMPK) are stimulated with globular and full-length Ad in skeletal muscle and only with full-length Ad in the liver. In parallel with its activation of AMPK, Ad stimulates phosphorylation of acetyl coenzyme A carboxylase (ACC), fatty-acid oxidation, glucose uptake and lactate production in myocytes, phosphorylation of ACC and reduction of molecules involved in gluconeogenesis in the liver, and reduction of glucose levels in vivo. Blocking AMPK activation by dominant-negative mutant inhibits each of these effects, indicating that stimulation of glucose utilization and fatty-acid oxidation by Ad occurs through activation of AMPK. Our data may provide a novel paradigm that an adipocyte-derived antidiabetic hormone, Ad, activates AMPK, thereby directly regulating glucose metabolism and insulin sensitivity in vitro and in vivo.
中文翻译：
Adiponectin是一个调控能源体内平衡和葡萄糖和脂质新陈代谢的脂肪细胞因子。 然而, 其调控能源体内平衡和葡萄糖和脂质新陈代谢具体的作用机制不是很清楚。在这里我们证明了“磷酸化作用”和“AMPK”在球状和全长的Adiponectin在肌肉中以及全长Adiponectin在肝脏中的激活起着重要作用。 在激活 AMPK的同时, Adiponectin刺激了乙酰辅酶A羧化酶的磷酸化、脂肪酸氧化 、葡萄糖摄取和乳酸盐生成。乙酰辅酶A羧化酶的磷酸化和葡萄糖异生的产生与、又导致血糖的下降。阻断了AMPK的作用之后的结果显示了Adiponectin的促脂肪氧化 、葡萄糖利用增加作用均是ampk介导的。我们的实验在体内和体外均证实了这样一个新的范例。
个人感慨：这篇文章的内容我并不想多讲，是做一个蛋白的信号转导的。大家知道，日 本 人现在在国际顶尖的“CNS”杂志上发表文章很多，特别是蛋白做得很好，那么，大家可以看看他们做这篇文章用到了什么技术
我列举一下：
1、Generation of recombinant Ad. 蛋白的重组表达
2、Western-blot analysis
3、RNA preparation and northern-blot analysis.
4、Lipid and glucose metabolism. 属于最简单的生化指标（血糖血脂）测定
5、Studies with C2C12 myocytes and isolated hepatocytes
6、Adenovirus-mediated gene transfer in vivo and Ad tolerance test.
7、Measurement of AMP, ADP and ATP content. 放免法测定
各位战友看看，这些技术难不难？可能5、6项还算是有点难度的，而其他的工作是不是我们蛋白版里天天讨论的那些技术？日 本 人就是能善于利用普通技术，如Western，Northern，免疫组化，elisa这些，组合起来运用说明一个问题。选择的切入点、实验设计的巧妙性，这些真是我们要好好学习的。人家有这些技术为什么就能发Nature，而我们却还在为一两分的SCI苦苦拼搏。。值得深思啊
文章我一同附上了 大家看看 全是Western的图片。。。
