Proteomic analysis for the anti-apoptotic effects of cystamine on apoptosis-prone macrophage.
Increased macrophage vulnerability is associated with progression of systemic lupus erythematosus. Our previous studies have shown that cystamine, an inhibitor of transglutaminase 2 (TG2), alleviated the apoptosis of hepatocyte and brain cell in lupus-prone mice NZB/W-F1. In present study, we further investigated the effects of cystamine on apoptosis-prone macrophages (APMs) in the lupus mice. Using two-dimensional gel electrophoresis (2-DE) analysis, we found that cystamine induced a differential protein expression pattern of APM as comparing to the PBS control. The protein spots presenting differential level between cystamine and PBS treatment were then identified by peptide-mass fingerprinting (PMF). After bioinformatic analysis, these identified proteins were found involved in mitochondrial apoptotic pathway, oxidative stress, and mitogen-activated protein (MAP) kinase-mediated pathway. Further investigation revealed that cystamine significantly decreased the levels of apoptotic Bax and Apaf-1 and the activity of caspase-3, and increased the levels of anti-apoptotic Bcl-2 in APM. We also found that these apoptotic mediators were up-regulated in a correlation with the progression of lupus severity in NZB/W-F1, which were little affected in BALB/c mice. We also found that the reduced serum glutathione was restored by cystamine in NZB/W-F1. Interestingly, the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in APM and the phagocytic ability was diminished in presence of cystamine. In conclusion, our findings indicate that cystamine significantly inhibited mitochondrial pathway, induced antioxidant proteins, and diminished phosphorylation of extracellular ERK1/2, which may alleviate the apoptosis and the phagocytic ability of APM.
Kao
Shao-Hsuan
SH
Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan.
Hsu
Tsai-Ching
TC
Yu
Jau-Song
JS
Chen
Jeng-Ting
JT
Li
Sin-Lun
SL
Lai
Wen-Xian
WX
Tzang
Bor-Show
BS
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D003538:Cystamine / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation; D015151:Immunoblotting; D008180:Lupus Erythematosus, Systemic / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D008814:Mice, Inbred NZB; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods
D004791:Enzyme Inhibitors; D003538:Cystamine