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Title: Zinc protoporphyrin inhibition of lipopolysaccharide-, lipoteichoic acid-, and peptidoglycan-induced nitric oxide production through stimulating iNOS protein ubiquitination

Abstract

In the present study, zinc protoporphyrin (ZnPP), but not ferric protoporphyrin (FePP), tin protoporphyrin (SnPP), or zinc chloride (ZnCl{sub 2}), at the doses of 0.5, 1, and 2 {mu}M, dose-dependently inhibited lipopolysaccharide- (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN)-induced inducible nitric oxide (iNOS) and nitric oxide (NO) production with an increase in heme oxygenase 1 (HO-1) protein in RAW264.7 macrophages in a serum-free condition. NO inhibition and HO-1 induction by ZnPP were blocked by the separate addition of fetal bovine serum (FBS) and bovine serum albumin (BSA). A decrease in the iNOS/NO ratio and an increase in HO-1 protein by ZnPP were identified in three different conditions including ZnPP pretreatment, ZnPP co-treatment, and ZnPP post-treatment with LPS and LTA. Activation of c-Jun N-terminal kinases (JNKs) and extracellular regulated kinases (ERKs) were detected in LPS-, LTA-, and PGN-treated RAW264.7 cells, and iNOS/NO production was blocked by adding the JNK inhibitor, SP600125, but not the ERK inhibitor, PD98059. However, ZnPP addition potentiated ERK and JNK protein phosphorylation stimulated by LPS, LTA, and PGN. Increases in total protein ubiquitination and ubiquitinated iNOS proteins were detected in ZnPP-treated macrophages elicited by LPS according to Western and immunoprecipitation/Western blotting assays, respectively. The decrease in LPS-inducedmore » iNOS protein by ZnPP was reversed by adding the proteasome inhibitors MG132 and lactacystin. The reduction in HO-1 protein induced by ZnPP via transfection of HO-1 small interfering RNA did not affect the inhibitory effect of ZnPP against LPS-induced iNOS/NO production and protein ubiquitination induced by ZnPP in macrophages. Data of the present study provide the first evidence to support ZnPP effectively inhibiting inflammatory iNOS/NO production through activation of protein ubiquitination in a HO-1-independent manner in macrophages.« less

Authors:
 [1]; ;  [2];  [3];  [4];  [5];  [6];  [2];  [7]
  1. Section of Hematology-Oncology, Department of Internal Medicine, Taipei Municipal Wan Fang Hospital, Taipei Medical University, Taipei 110, Taiwan (China)
  2. Graduate Institute of Medical Sciences, Taipei Medical University, Taipei 110, Taiwan (China)
  3. Department of Gastroenterology, Taipei Medical University Wan Fang Hospital, Taiwan (China)
  4. Graduate Institute of Pharmacy, School of Pharmacy, Taipei Medical University, Taipei 110, Taiwan (China)
  5. Department of Neurosurgery, Taipei Medical University-Wan Fang Hospital, Taipei, Taiwan (China)
  6. Graduate Institute of Medical Sciences, Taipei Medical University, Taipei 110, Taiwan (China), E-mail: chlin@tmu.edu.tw
  7. (China), E-mail: yc3270@tmu.edu.tw
Publication Date:
OSTI Identifier:
21272584
Resource Type:
Journal Article
Journal Name:
Toxicology and Applied Pharmacology
Additional Journal Information:
Journal Volume: 237; Journal Issue: 3; Other Information: DOI: 10.1016/j.taap.2009.04.009; PII: S0041-008X(09)00159-8; Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0041-008X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ALBUMINS; HEME; INFLAMMATION; INHIBITION; MACROPHAGES; NITRIC OXIDE; PHOSPHORYLATION; PHOSPHOTRANSFERASES; RNA; TIN; ZINC; ZINC CHLORIDES

Citation Formats

Chow, J.-M., Lin, H.-Y., Shen, S.-C., Wu, M.-S., Lin, C.-W., Chiu, W.-T., Lin, C.-H., Chen, Y.-C., and Cancer Research Center and Orthopedics Research Center, Taipei Medical University Hospital, Taipei, Taiwan. Zinc protoporphyrin inhibition of lipopolysaccharide-, lipoteichoic acid-, and peptidoglycan-induced nitric oxide production through stimulating iNOS protein ubiquitination. United States: N. p., 2009. Web. doi:10.1016/j.taap.2009.04.009.
Chow, J.-M., Lin, H.-Y., Shen, S.-C., Wu, M.-S., Lin, C.-W., Chiu, W.-T., Lin, C.-H., Chen, Y.-C., & Cancer Research Center and Orthopedics Research Center, Taipei Medical University Hospital, Taipei, Taiwan. Zinc protoporphyrin inhibition of lipopolysaccharide-, lipoteichoic acid-, and peptidoglycan-induced nitric oxide production through stimulating iNOS protein ubiquitination. United States. doi:10.1016/j.taap.2009.04.009.
Chow, J.-M., Lin, H.-Y., Shen, S.-C., Wu, M.-S., Lin, C.-W., Chiu, W.-T., Lin, C.-H., Chen, Y.-C., and Cancer Research Center and Orthopedics Research Center, Taipei Medical University Hospital, Taipei, Taiwan. Mon . "Zinc protoporphyrin inhibition of lipopolysaccharide-, lipoteichoic acid-, and peptidoglycan-induced nitric oxide production through stimulating iNOS protein ubiquitination". United States. doi:10.1016/j.taap.2009.04.009.
@article{osti_21272584,
title = {Zinc protoporphyrin inhibition of lipopolysaccharide-, lipoteichoic acid-, and peptidoglycan-induced nitric oxide production through stimulating iNOS protein ubiquitination},
author = {Chow, J.-M. and Lin, H.-Y. and Shen, S.-C. and Wu, M.-S. and Lin, C.-W. and Chiu, W.-T. and Lin, C.-H. and Chen, Y.-C. and Cancer Research Center and Orthopedics Research Center, Taipei Medical University Hospital, Taipei, Taiwan},
abstractNote = {In the present study, zinc protoporphyrin (ZnPP), but not ferric protoporphyrin (FePP), tin protoporphyrin (SnPP), or zinc chloride (ZnCl{sub 2}), at the doses of 0.5, 1, and 2 {mu}M, dose-dependently inhibited lipopolysaccharide- (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN)-induced inducible nitric oxide (iNOS) and nitric oxide (NO) production with an increase in heme oxygenase 1 (HO-1) protein in RAW264.7 macrophages in a serum-free condition. NO inhibition and HO-1 induction by ZnPP were blocked by the separate addition of fetal bovine serum (FBS) and bovine serum albumin (BSA). A decrease in the iNOS/NO ratio and an increase in HO-1 protein by ZnPP were identified in three different conditions including ZnPP pretreatment, ZnPP co-treatment, and ZnPP post-treatment with LPS and LTA. Activation of c-Jun N-terminal kinases (JNKs) and extracellular regulated kinases (ERKs) were detected in LPS-, LTA-, and PGN-treated RAW264.7 cells, and iNOS/NO production was blocked by adding the JNK inhibitor, SP600125, but not the ERK inhibitor, PD98059. However, ZnPP addition potentiated ERK and JNK protein phosphorylation stimulated by LPS, LTA, and PGN. Increases in total protein ubiquitination and ubiquitinated iNOS proteins were detected in ZnPP-treated macrophages elicited by LPS according to Western and immunoprecipitation/Western blotting assays, respectively. The decrease in LPS-induced iNOS protein by ZnPP was reversed by adding the proteasome inhibitors MG132 and lactacystin. The reduction in HO-1 protein induced by ZnPP via transfection of HO-1 small interfering RNA did not affect the inhibitory effect of ZnPP against LPS-induced iNOS/NO production and protein ubiquitination induced by ZnPP in macrophages. Data of the present study provide the first evidence to support ZnPP effectively inhibiting inflammatory iNOS/NO production through activation of protein ubiquitination in a HO-1-independent manner in macrophages.},
doi = {10.1016/j.taap.2009.04.009},
journal = {Toxicology and Applied Pharmacology},
issn = {0041-008X},
number = 3,
volume = 237,
place = {United States},
year = {2009},
month = {6}
}