Lee, Ji Ae Kwon, Young-Won Kim, Hye Ri Shin, Nari Son, Hyo Jin Cheong, Chan Seong Kim, Dong Jin Hwang, Onyou 2024-01-19T12:32:24Z 2024-01-19T12:32:24Z 2022-04-03 2022-03 1016-8478 https://pubs.kist.re.kr/handle/201004/115583 The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin-1 beta (IL-1 beta), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-beta [TGF-beta]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress- and inflammation-related neurodegenerative disorders such as Parkinson's disease. English KOREAN SOC MOLECULAR & CELLULAR BIOLOGY A Novel Pyrazolo[3,4-d]pyrimidine Induces Heme Oxygenase-1 and Exerts Anti-Inflammatory and Neuroprotective Effects Article 10.14348/molcells.2021.0074 1 MOLECULES AND CELLS, v.45, no.3, pp.134 - 147 MOLECULES AND CELLS 45 3 134 147 N scie scopus kci ART002828395 000742809000001 Biochemistry & Molecular Biology Cell Biology Biochemistry & Molecular Biology Cell Biology Article; Early Access ACTIVATED PROTEIN-KINASE NF-KAPPA-B DOPAMINERGIC-NEURONS ENDOTHELIAL-CELLS UPSTREAM KINASE AMPK NRF2 MICROGLIA BETA INFLAMMATION heme oxygenase-1 neuroinflammation Parkinson&apos s disease pyrazolo[3,4-d]pyrimidine