PMID- 19781192 OWN - NLM STAT- MEDLINE DCOM- 20110324 LR - 20171116 IS - 0529-5807 (Print) IS - 0529-5807 (Linking) VI - 38 IP - 7 DP - 2009 Jul TI - [NADPH oxidase-derived reactive oxygen species involved in angiotensin II-induced monocyte chemoattractant protein-1 expression in mesangial cells]. PG - 456-61 AB - OBJECTIVE: To investigate the origin of oxidative stress induced by angiotensin II (AngII) in human mesangial cells and the role of reactive oxygen species (ROS) in AngII-induced monocyte chemoattractant protein-1 (MCP-1) expression. METHODS: MCP-1 expression was determined by real time RT-PCR. ROS production was measured by DCFDA fluorescence. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was examined by lucigenin chemiluminescence. p47phox and p67phox translocation was assayed by Western blot. Twenty-four male mice were randomly divided into three groups: the control, the AngIIinfusion [AngII 400 ng/(kg.min)], and the apocynin treatment. AngII was infused by subcutaneously osmotic minipump for 14 days. Urinary albumin and 8-isoprostane excretion were measured by ELISA. RESULTS: In cultured human mesangial cells, AngII induced the MCP-1 expression in a dose-dependent manner with 3.56 fold increase as compared with the control. AngII increased intracellular ROS production as early as 3 min with the peak at 60 min and was in a time and dose-dependent. Incubation with different dosages of AngII (1 nmol/L, 10 nmol/L, and 100 nmol/L AngII) for 60 min, ROS production increased at 1.82, 2.92, and 4.08 folds respectively. AngII-induced ROS generation was sensitive to diphenyleneiodonium sulfate (DPI, 10 micromol/L) and apocynin (500 micromol/L), two structurally distinct NADPH oxidase inhibitors. In contrast, inhibitors of other oxidant-producing enzymes, including the mitochondrial complex Iinhibitor rotenone, the xanthine oxidase inhibitor allopurinol, the cyclooxygenase inhibitor indomethacin, the lipoxygenase inhibitor nordihydroguiaretic acid, the cytochrome P450 oxygenase inhibitor ketoconazole and the nitric oxide synthase inhibitor G-nitro-L-arginine methyl ester were without an effect. AngII-induced ROS generation was inhibited by the AT1 antagonist losartan (10 micromol/L) but not the AT2 antagonist PD123319 (10 micromol/L). AngII treatment induced translocation of cytosolic of p47phox and p67phox to the membrane. The antioxidants almost abolished AngII-induced MCP-1 expression. AngII infusion increased urinary and p67 translocation by 2.69-, 2.97-, and 2.67-fold, respectively. CONCLUSIONS: NADPH oxidase-derived ROS is involved in AngII-induced MCP-1 expression. Inhibition of NADPH oxidase alleviates AngII-induced renal injury. FAU - Chen, Ying AU - Chen Y AD - Department of Nephrology, Nanjing Children's Hospital, Affiliated to Nanjing Medical University, Nanjing 210008, China. FAU - Zhang, Ai-hua AU - Zhang AH FAU - Huang, Song-ming AU - Huang SM FAU - Ding, Gui-xia AU - Ding GX FAU - Zhang, Wei-zhen AU - Zhang WZ FAU - Bao, Hua-ying AU - Bao HY FAU - Wu, Hong-mei AU - Wu HM FAU - Chen, Rong-hua AU - Chen RH LA - chi PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - China TA - Zhonghua Bing Li Xue Za Zhi JT - Zhonghua bing li xue za zhi = Chinese journal of pathology JID - 0005331 RN - 0 (Acetophenones) RN - 0 (Angiotensin II Type 1 Receptor Blockers) RN - 0 (CCL2 protein, human) RN - 0 (Chemokine CCL2) RN - 0 (Onium Compounds) RN - 0 (Phosphoproteins) RN - 0 (Reactive Oxygen Species) RN - 0 (neutrophil cytosol factor 67K) RN - 11128-99-7 (Angiotensin II) RN - 6HJ411TU98 (diphenyleneiodonium) RN - B6J7B9UDTR (acetovanillone) RN - EC 1.6.3.- (NADPH Oxidases) RN - EC 1.6.3.1 (neutrophil cytosolic factor 1) RN - JMS50MPO89 (Losartan) SB - IM MH - Acetophenones/pharmacology MH - Angiotensin II/administration & dosage/*pharmacology MH - Angiotensin II Type 1 Receptor Blockers/pharmacology MH - Animals MH - Cells, Cultured MH - Chemokine CCL2/*metabolism MH - Dose-Response Relationship, Drug MH - Humans MH - Losartan/pharmacology MH - Male MH - Mesangial Cells/*metabolism MH - Mice MH - Mice, Inbred C57BL MH - NADPH Oxidases/antagonists & inhibitors/*metabolism MH - Onium Compounds/pharmacology MH - Oxidative Stress MH - Phosphoproteins/metabolism MH - Protein Transport MH - Random Allocation MH - Reactive Oxygen Species/*metabolism EDAT- 2009/09/29 06:00 MHDA- 2011/03/25 06:00 CRDT- 2009/09/29 06:00 PHST- 2009/09/29 06:00 [entrez] PHST- 2009/09/29 06:00 [pubmed] PHST- 2011/03/25 06:00 [medline] PST - ppublish SO - Zhonghua Bing Li Xue Za Zhi. 2009 Jul;38(7):456-61.