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Chlorophyllin suppression of lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells

Authors
 Kyung-Joo Cho  ;  Seung Hyun Han  ;  Bu Yeo Kim  ;  Seong-Gu Hwang  ;  Kwang-Kyun Park  ;  Kyu-Hwan Yang  ;  An-Sik Chung 
Citation
 Toxicology and Applied Pharmacology, Vol.166(2) : 120-127, 2000 
Journal Title
TOXICOLOGY AND APPLIED PHARMACOLOGY
ISSN
 0041-008X 
Issue Date
2000
MeSH
Animals ; Antimutagenic Agents/pharmacology* ; Cell Line ; Cell Nucleus/metabolism ; Chloramphenicol O-Acetyltransferase/genetics ; Chlorophyllides/pharmacology* ; DNA/metabolism ; DNA-Binding Proteins/drug effects ; DNA-Binding Proteins/metabolism ; Dose-Response Relationship, Drug ; Electrophoresis ; Enzyme Inhibitors/pharmacology* ; Genes, Reporter ; Lipopolysaccharides/pharmacology ; Lymphocyte Activation ; Macrophages/drug effects* ; Macrophages/enzymology ; Mice ; Mice, Inbred BALB C ; NF-kappa B/genetics ; NF-kappa B/metabolism ; Nitric Oxide Synthase/antagonists & inhibitors* ; Nitric Oxide Synthase/genetics ; Nitric Oxide Synthase/metabolism ; Nitric Oxide Synthase Type II ; Nitrites/metabolism ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Salmonella typhimurium ; Transfection
Keywords
chlorophyllin ; nitric oxide ; lipopolysaccharide ; NF-κB
Abstract
Chlorophyllin (CHL), a water-soluble derivative of chlorophyll, functions as an anticarcinogen and antioxidant. In the present study, we investigated the effect of CHL on nitric oxide production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Treatment with CHL inhibited nitric oxide production in the LPS-stimulated RAW 264. 7 cells in a dose-related manner. Competitive RT-PCR analysis, using a DNA competitor as an internal standard, demonstrated that the treatment with 1, 10, and 50 microM CHL decreased LPS-induced iNOS mRNA expression in a concentration-dependent manner. Since the expression of the iNOS gene is mainly regulated by NF-kappaB, we then examined the effects of CHL on the NF-kappaB DNA binding activity, using an electrophoretic mobility shift assay. CHL down-regulated the NF-kappaB DNA binding on its cognate recognition site at the concentrations just noted. Employing a transfection and reporter gene expression system with p(NF-kappaB)(3)-chloramphenicol acetyl transferase (CAT), the treatment of CHL produced a dose-dependent inhibition of CAT activity in RAW 264.7 cells. Furthermore, CHL partially restored LPS-decreased IkappaBalpha, an inhibitory protein against NF-kappaB activation, in the cytosolic extract from the LPS-treated cells determined by immunoblot analysis. CHL also protected the hydroxyl radical-induced cytotoxicity in RAW 264.7 cells, indicating its antioxidant effect. These results suggest that CHL suppresses the nitric oxide production and iNOS mRNA expression mediated by the inhibition of NF-kappaB activation, and its action mechanism may be based on its antioxidant effect.
Full Text
https://www.sciencedirect.com/science/article/pii/S0041008X00989580
DOI
10.1006/taap.2000.8958
Appears in Collections:
2. College of Dentistry (치과대학) > Dept. of Oral Biology (구강생물학교실) > 1. Journal Papers
Yonsei Authors
Park, Kwang Kyun(박광균)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/171869
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