轉麩醯胺酸酶抑制劑對狼瘡鼠肝臟凋亡之影響及相關機制之探討

The study of transglutaminase2 inhibitor on the apoptosis and its related mechanisms on liver of NZB/W F1 mice

賴文賢(Wen-Xian Lai)

指導教授 ： 曾博修

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2007年)

https://doi.org/10.6834/CSMU.2007.00046

摘要

全身性紅斑性狼瘡為慢性自體免疫疾病，可能會影響肝臟而造成傷害，近年來有文獻指出Transglutaminase2(TG2)可能與全身性紅斑性狼瘡病理有關，因此本篇利用TG2抑制劑胱胺，利用NZB/W F1作為實驗動物模式，以探討胱胺在全身性紅斑性狼瘡肝臟所扮演的角色為何，由實驗結果發現胱胺可以降低NZB/W F1血清中的ALT及AST，而利用西方墨點法發現C反應蛋白表現在血清及肝臟也具有下降的情形，透過免疫組織染色法發現胱胺可以減少NZB/W F1肝臟alpha-SMA的表現，由TUNEL assay可以看到肝臟凋亡細胞的減少，利用西方墨點法看到胱胺在NZB/W F1肝臟可以降低與凋亡相關蛋白Fas、active caspase 8、Bad、cytochrome c、Apaf-1的表現量及增加Bcl-2蛋白表現量，與細胞週期相關的蛋白p53、p21也有降低的現象，而NF-kB p65蛋白表現量增加，此外與壓力相關蛋白Hsp70的表現量降低，因此在我們實驗結果發現胱胺可能透過抑制細胞凋亡，來對狼瘡小鼠NZB/W F1肝臟達到保護的作用。

關鍵字

轉麩醯胺酸酶紅斑性狼瘡 ； 細胞凋亡

並列摘要

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with increased production of autoantibodies. Recently, various studies have the increased population of liver abnormality in patients with SLE and associated Transglutaminase2(TG2) in the pathogenesis of SLE. In our purpose, to investigate the effects of transglutaminase inhibitor cystamine on liver of NZB/W F1 mice. We use that NZB/W F1 mice were used as the experimental animal model. We found that both ALT and AST were decreased in sera of NZB/W F1 mice treated with cystamine. The C-reactive protein expression was decreased in both sera and liver of NZB/W F1 mice treated with cystamine. Additionally, the alpha-SMA expression was detected by Immunohistochemistry. We found that cystamine can reduce alpha-SMA expression in NZB/W F1 liver. The apoptotic cell death of liver was detected by TUNEL assay. We found that cystamine can reduce apoptosis of liver in NZB/W F1. The expression of protein was detected by western blotting. The pro-apoptotic Fas, active caspase8, Bad, cytochrome c and Apaf-1 proteins were significantly decreased in liver of NZB/W F1 mice treated with cystamine. The anti-apoptotic Bcl-2 protein was significantly increased. The cell cycle proteins of p21and p53 were significantly decreased in the treated NZB/W F1 mice liver. The NF-kB p65 protein expression was increased. In addition, we observed decreased stress-related Hsp70 production in the treated NZB/W F1 mice liver. In our results show that cystamine can inhibit apoptosis and protect NZB/W F1 mice liver.

並列關鍵字

TG2 ； SLE ； apoptosis

參考文獻

20. Feng, H., Zhang, G. Y., Xie, H. F., Chen, M. L., Shi, W., and Wang, L. Y. [Hepatitis C virus infection combined with systemic lupus erythematosus]. Zhong Nan Da Xue Xue Bao Yi Xue Ban, 31: 891-893, 2006.

1. Croker, J. A. and Kimberly, R. P. SLE: challenges and candidates in human disease. Trends Immunol, 26: 580-586, 2005.

2. D'Cruz, D. P., Khamashta, M. A., and Hughes, G. R. Systemic lupus erythematosus. Lancet, 369: 587-596, 2007.

3. Singh, R. R. SLE: translating lessons from model systems to human disease. Trends Immunol, 26: 572-579, 2005.

4. Banchereau, J. and Pascual, V. Type I interferon in systemic lupus erythematosus and other autoimmune diseases. Immunity, 25: 383-392, 2006.