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- Title
Role of NAD(P)H oxidase in transforming growth factor-β1-induced monocyte chemoattractant protein-1 and interleukin-6 expression in rat renal tubular epithelial cells.
- Authors
HAIYAN ZHANG; ZONGPEI JIANG; JIE CHANG; XIAOYAN LI; HENGMEI ZHU; HUI Y LAN; SHU-FENG ZHOU; XUEQING YU
- Abstract
Aim: This study investigated the role of NAD(P)H oxidase in transforming growth factor-β1 (TGF-β1)-induced reactive oxygen species (ROS) generation, monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6) expression in rat renal tubular epithelial NRK-52E cells. Methods: The cells were treated with 10 ng/mL TGF-β1, either in the presence or absence of the NAD(P)H oxidase inhibitor, diphenyleneiodonium (DPI), or short hairpin RNA (shRNA) suppressing p67phox expression. Expression of NAD(P)H oxidase subunits, MCP-1, and IL-6 at the mRNA levels was detected by reverse transcription polymerase chain reaction, while expression of NAD(P)H oxidase subunit p67phox protein was analyzed by western blot and MCP-1 by enzyme-linked immunosorbent assay. The cellular ROS generation was visualized using 2′,7′-dichlorodihydrofluorescein diacetate by confocal microscopy. Results: Compared to control, TGF-β1 upregulated NAD(P)H oxidase subunit p67phox mRNA by 3.59-fold ( P < 0.01), but had no effect on p22phox, gp91phox and p47phox NAD(P)H subunits. TGF-β1 was also able to significantly increase intracellular ROS ( P < 0.05), MCP-1 ( P < 0.01) and IL-6 ( P < 0.05) expression in NRK-52E cells. Further studies showed that generation of ROS and upregulation of MCP-1 and IL-6 by TGF-β1 were significantly blocked by addition of DPI or shRNA-p67phox ( P < 0.01), suggesting that these effects were NAD(P)H oxidase-dependent. Conclusion: TGF-β1 differentially regulates the expression of NAD(P)H oxidase subunits and mediates MCP-1 and IL-6 expression in rat renal tubular cells via the NAD(P)H oxidase/p67phox-dependent mechanism.
- Subjects
OXIDASES; MONOCYTES; RENAL tubular transport; EPITHELIAL cells; POLYMERASE chain reaction
- Publication
Nephrology, 2009, Vol 14, Issue 3, p302
- ISSN
1320-5358
- Publication type
Article
- DOI
10.1111/j.1440-1797.2008.01072.x