The Effect of FK506 to Generate Reactive Oxygen Species on T Lymphocyte Death.
10.4174/jkss.2009.77.5.310
- Author:
Ho Kyun LEE
1
;
Sang Young CHUNG
;
Soo Jin CHOI
Author Information
1. Department of Surgery, Chonnam National University Medical School, Gwangju, Korea. choisjn@chonnam.ac.kr
- Publication Type:Original Article
- Keywords:
FK506;
Reactive oxygen species;
Heme Oxygenase-1
- MeSH:
Apoptosis;
Blotting, Western;
Flow Cytometry;
Fluorescence;
Heme;
Heme Oxygenase-1;
Humans;
Hydrogen Peroxide;
Immunohistochemistry;
Indoles;
Jurkat Cells;
Lymphocytes;
Oxidative Stress;
Proteins;
Puma;
Reactive Oxygen Species;
Rejection (Psychology);
Superoxides;
T-Lymphocytes;
Tacrolimus;
Transplants
- From:Journal of the Korean Surgical Society
2009;77(5):310-319
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: Tacrolimus (FK506) has been widely used as an immunosuppressant in organ transplanted recipients to suppress organ rejection phenomenon. We investigated the role of oxidative stress and heme oxygense-1 by FK506 on human Jurkat T cells. METHODS: The cells viability was examined by DAPI stain, enzyme activity of caspase family proteins, and western blotting for Baks, PUMA, iNOS, HO-1. Cells were cultured in the absence or presence of CoPPIX or ZnPPIX and the fluorescence intensity was analyzed using a flow cytometry. RESULTS: Treatment with FK506 increased the generation of reactive oxygen species (ROS), including hydrogen peroxide and superoxide anion, and NO in Jurkat cells in a dose-dependent manner. Immunohistochemistry and Western blot analysis data revealed the hemoxygenase-1 (HO-1) was induced by the addition of FK506 in Jurkat cells. Induction of CoPP, HO-1 inducer, resulted in decreased intracellular H2O2 and NO concentrations. Instead ZnPP, an HO-1 competitive inhibitor did it reversely. In addition, ZnPP regulates iNOS protein synthesis by inhibition of HO-1. CONCLUSION: Increase of HO-1 expression would induce to decrease the intracellular H2O2 and NO concentrations. Also, HO-1 would regulate iNOS protein synthesis. Consequently, we can expect the regulation of HO-1 expression with concomitants use of FK506 to suppress organ rejection phenomenon by enhancing apoptosis.
