title: Andrographolide inhibits TNFα-induced ICAM-1 expression via suppression of NADPH oxidase activation and induction of HO-1 and GCLM expression through the PI3K/Akt/Nrf2 and PI3K/Akt/AP-1 pathways in human endothelial cells.
abstract: Andrographolide, the major bioactive component of Andrographis paniculata, has been demonstrated to have various biological properties including anti-inflammation, antioxidation, and anti-hepatotoxicity. Oxidative stress is considered a major risk factor in aging, inflammation, cancer, atherosclerosis, and diabetes mellitus. NADPH oxidase is a major source of endogenous reactive oxygen species (ROS). In this study, we used EA.hy926 endothelial-like cells to explore the anti-inflammatory activity of andrographolide. Andrographolide attenuated TNFα-induced ROS generation, Src phosphorylation, membrane translocation of the NADPH oxidase subunits p47(phox) and p67(phox), and ICAM-1 gene expression. In the small hairpin RNA interference assay, shp47(phox) abolished TNFα-induced p65 nuclear translocation, ICAM-1 gene expression, and adhesion of HL-60 cells. Andrographolide induced the gene expression of heme oxygenase 1 (HO-1) and glutamate cysteine ligase modifier subunit (GCLM) in a time-dependent manner. Cellular glutathione (GSH) content was increased by andrographolide. shGCLM attenuated the andrographolide-induced increase in GSH content and reversed the andrographolide inhibition of HL-60 adhesion. shHO-1 showed a similar effect on andrographolide inhibition of HL-60 adhesion to shGCLM. The mechanism underlying the up-regulation of HO-1 and GCLM by andrographolide was dependent on the PI3K/Akt pathway, and both the Nrf2 and AP-1 transcriptional factors were involved. Our results suggest that andrographolide attenuates TNFα-induced ICAM-1 expression at least partially through suppression of NADPH oxidase activation and induction of HO-1 and GCLM expression, which is PI3K/Akt pathway-dependent.
authors.affiliation: Department of Nutrition, China Medical University, Taichung 404, Taiwan.
authors.forename: Chia-Yang
authors.initials: CY
authors.lastname: Lu
authors.affiliation: Department of Health and Nutrition Biotechnology, Asia University, Taichung 413, Taiwan.
authors.forename: Ya-Chen
authors.initials: YC
authors.lastname: Yang
authors.affiliation: School of Nutrition, Chung Shan Medical University, Taichung 402, Taiwan; Department of Nutrition, Chung Shan Medical University Hospital, Taichung 402, Taiwan.
authors.forename: Chien-Chun
authors.initials: CC
authors.lastname: Li
authors.affiliation: School of Nutrition, Chung Shan Medical University, Taichung 402, Taiwan; Department of Nutrition, Chung Shan Medical University Hospital, Taichung 402, Taiwan.
authors.forename: Kai-Li
authors.initials: KL
authors.lastname: Liu
authors.affiliation: Department of Nutrition, China Medical University, Taichung 404, Taiwan; Department of Health and Nutrition Biotechnology, Asia University, Taichung 413, Taiwan. Electronic address: cklii@mail.cmu.edu.tw.
authors.forename: Chong-Kuei
authors.initials: CK
authors.lastname: Lii
authors.affiliation: Department of Nutrition, China Medical University, Taichung 404, Taiwan. Electronic address: chenhw@mail.cmu.edu.tw.
authors.forename: Haw-Wen
authors.initials: HW
authors.lastname: Chen
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D004224:Diterpenes / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D005721:Glutamate-Cysteine Ligase / Q000235:genetics* / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D018922:HL-60 Cells / Q000187:drug effects; D051547:Heme Oxygenase-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism*; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D019869:Phosphatidylinositol 3-Kinases; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000494:pharmacology
chemical_list: D004224:Diterpenes; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione; D051547:Heme Oxygenase-1; D018799:Intercellular Adhesion Molecule-1; D048031:JNK Mitogen-Activated Protein Kinases; D019255:NADPH Oxidases; D051267:NF-E2-Related Factor 2; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D017382:Reactive Oxygen Species; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; C495635:NFE2L2 protein, human; C030419:andrographolide; C496541:HMOX1 protein, human; C546365:GCLM protein, human
keywords: Activator protein 1 (AP-1); Glutamate cysteine ligase modifier subunit (GCLM); Heme oxygenase 1 (HO-1); NADPH oxidase; Nuclear factor erythroid 2-related factor 2 (Nrf2)