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ACTIVATION OF NRF2 COORDINATES DDAH/PPAR-?/eNOS PATHWAYS THAT ENHANCE NITRIC OXIDE GENERATION IN HUMAN GLOMERULAR ENDOTHELIAL CELLS #MMPMID25691623
Luo Z; Aslam S; Welch WJ; Wilcox CS
Hypertension 2015[Apr]; 65 (4): 896-902 PMID25691623show ga
Dimethylarginine dimethylaminohydrolase (DDAH) degrades (ADMA) which inhibits nitric oxide synthase (NOS). Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcriptional factor that binds to antioxidant response elements (ARE) and transcribes many antioxidant genes. Since the promoters of the human DDAH-1 and -2, eNOS and PPAR- ? genes contain 2?3 putative AREs, we hypothesized that they were regulated by Nrf2/ARE. Incubation of human renal glomerular endothelial cells (HRGECs) with the Nrf2 activator tert-butylhydroquinone (tBHQ) (20 µmol?l?1) significantly (P<0.05) increased NO and activities of NOS and DDAH and decreased ADMA. It upregulated genes for hemoxygenase -1, eNOS, DDAH-1 and -2 and PPAR-? and partitioned Nrf2 into the nucleus. Knockdown of Nrf2 abolished these effects. Nrf2 bound to one ARE on the DDAH-1 and -2 and PPAR-? promoters but not to the eNOS promoter. An increased eNOS and phosphorylated eNOS (P-eNOSser-1177) expression with tBHQ was prevented by knockdown of PPAR-?. Expression of Nrf2 was reduced by knockdown of PPAR-? whereas PPAR-? was reduced by knockdown of Nrf2, thereby demonstrating 2-way positive interactions. Thus, Nrf2 transcribes the HO-1 and other genes to reduce reactive oxygen species, and DDAH-1 and -2 to reduce ADMA and PPAR-? to increase eNOS and its phosphorylation and activity thereby coordinating three pathways that enhance endothelial NO generation.