Increased phagocyte-like NADPH oxidase and ROS generation in type 2 diabetic ZDF rat and human islets: role of Rac1-JNK1/2 signaling pathway in mitochondrial dysregulation in the diabetic islet.
Increased phagocyte-like NADPH oxidase and ROS generation in type 2 diabetic ZDF rat and human islets: role of Rac1-JNK1/2 signaling pathway in mitochondrial dysregulation in the diabetic islet.
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作者：
Syed I;Kyathanahalli CN;Jayaram B;Govind S;Rhodes CJ;Kowluru RA;Kowluru A
确定Zucker糖尿病脂肪（ZDF）大鼠和糖尿病人胰岛中吞噬细胞样NADPH氧化酶（Nox）的亚基表达和功能激活、活性氧（ROS）生成和caspase-3激活。通过Western印迹和光密度法定量Nox核心组分的表达。用2′，7 ′-二氯荧光素二乙酸酯法定量测定ROS水平。Rac 1激活使用金标记的免疫吸附测定试剂盒进行定量。磷酸化p47 phox，活性Rac 1，Nox活性，ROS生成，Jun NH 2-末端激酶（JNK）1/2磷酸化和caspase-3活性的水平在ZDF胰岛中显著高于瘦对照大鼠胰岛。INS 832/13细胞长期暴露于糖脂毒性条件导致JNK 1/2磷酸化和caspase-3活性增加;这种作用在很大程度上被JNK的选择性抑制剂SP 600125逆转。正常人胰岛与高糖孵育也增加了Rac 1和Nox的激活。最后，以类似于ZDF糖尿病大鼠胰岛的方式，Rac 1表达、JNK 1/2和caspase-3活化在糖尿病人胰岛中也显著增加。我们提供了第一个体外和体内证据，支持在胰岛β细胞中加速的Rac 1-Nox-ROS-JNK 1/2信号通路导致糖尿病中线粒体失调的发生。
To determine the subunit expression and functional activation of phagocyte-like NADPH oxidase (Nox), reactive oxygen species (ROS) generation and caspase-3 activation in the Zucker diabetic fatty (ZDF) rat and diabetic human islets. Expression of core components of Nox was quantitated by Western blotting and densitometry. ROS levels were quantitated by the 2′,7′-dichlorofluorescein diacetate method. Rac1 activation was quantitated using the gold-labeled immunosorbent assay kit. Levels of phosphorylated p47phox, active Rac1, Nox activity, ROS generation, Jun NH2-terminal kinase (JNK) 1/2 phosphorylation, and caspase-3 activity were significantly higher in the ZDF islets than the lean control rat islets. Chronic exposure of INS 832/13 cells to glucolipotoxic conditions resulted in increased JNK1/2 phosphorylation and caspase-3 activity; such effects were largely reversed by SP600125, a selective inhibitor of JNK. Incubation of normal human islets with high glucose also increased the activation of Rac1 and Nox. Lastly, in a manner akin to the ZDF diabetic rat islets, Rac1 expression, JNK1/2, and caspase-3 activation were also significantly increased in diabetic human islets. We provide the first in vitro and in vivo evidence in support of an accelerated Rac1–Nox–ROS–JNK1/2 signaling pathway in the islet β-cell leading to the onset of mitochondrial dysregulation in diabetes.
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影响因子：
7.7
作者：
MACDONALD, MJ
通讯作者：
MACDONALD, MJ
影响因子：
7.7
作者：
Kowluru, Anjaneyulu;Veluthakal, Rajakrishnan;Koch, Brandon J.
通讯作者：
Koch, Brandon J.
影响因子：
4.8
作者：
Shen, YH;Godlewski, J;Tzivion, G
通讯作者：
Tzivion, G
影响因子：
4.8
作者：
Morgan, D.;Rebelato, E.;Carpinelli, A. R.
通讯作者：
Carpinelli, A. R.
DOI：
10.1016/j.bbrc.2005.05.065
发表时间：
2005-07-15
影响因子：
3.1
作者：
Nakayama, M;Inoguchi, T;Nawata, H
通讯作者：
Nawata, H