High glucose condition upregulated Txnip expression level in rat mesangial cells through ROS/MEK/MAPK pathway
High glucose condition upregulated Txnip expression level in rat mesangial cells through ROS/MEK/MAPK pathway
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DOI：
10.1007/s11010-010-0626-z
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发表时间：
2011-01-01
影响因子：
4.3
通讯作者：
Zhou, Hongbo
中科院分区：
文献类型：
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作者：
Fang, Shaohong;Jin, Yuhong;Zhou, Hongbo
硫氧还蛋白相互作用蛋白（Txnip）是高血糖反应中表达上调最多的基因之一。在链脲佐菌素诱导的糖尿病小鼠中，Txnip的肾脏表达增加与IV型胶原积累相关。由于高糖的作用机制尚不清楚，我们对高糖诱导的大鼠系膜细胞Txnip表达上调的信号通路进行了研究。大鼠肾小球系膜细胞暴露于正常（5.5 mM）或高（25 mM）葡萄糖在不同的时间点。分别在转录和翻译水平上使用实时RT-PCR和蛋白质印迹法测定Txnip的表达。结果：高糖处理后4 ~ 12 h，细胞内Txnip mRNA表达增加，12 ~ 24 h，Txnip蛋白表达增加。此外，发现N-乙酰半胱氨酸（NAC）在高糖条件下降低Txnip蛋白的表达。此外，p38 MAPK抑制剂SB 203580在转录和蛋白水平上显着抑制高糖暴露下Txnip的表达。这些结果表明高糖暴露通过ROS/MEK/MAPK信号通路提高Txnip mRNA和蛋白的表达水平。
Thioredoxin interacting protein (Txnip) is one of the most abundantly up-regulated genes in response to hyperglycemia. The increased renal expression of Txnip was associated with type IV collagen accumulation in streptozotocin-induced diabetic mice. As the mechanism of action of high glucose is unknown, we undertook the investigation of the signaling pathway on the upregulation of Txnip expression induced by high glucose in rat mesangial cells. Rat mesangial cells were exposed to normal (5.5 mM) or high (25 mM) glucose at different time points. Txnip expression was determined using real-time RT-PCR and western-blotting at transcription and translation level, respectively. Intracellular reactive oxygen species (ROS) was detected by FACS Calibur flow cytometer using fluorescent probe (DCFH-DA).The treatment with high glucose resulted in an increase of Txnip mRNA from 4 h to 12 h and Txnip protein from 12 to 24 h in comparison with normal glucose condition. In addition, N-acetyl-cysteine (NAC) was found to decrease Txnip protein expression under high glucose condition. Furthermore, p38MAPK inhibitor SB203580 suppressed Txnip expression at transcription and protein level significantly to high glucose exposure. These results suggest that high glucose exposure improves Txnip mRNA and protein expression level by ROS/MEK/MAPK signaling pathway.