Tudan Christopher, Jackson John K, Higo Tobi T, Hampong Maggie, Pelech Steven L, Burt Helen M
Faculty of Pharmaceutical Sciences, 2146 East Mall, University of British Columbia, Vancouver, BC, Canada V6T 1Z3.
Cell Signal. 2004 Feb;16(2):211-21. doi: 10.1016/j.cellsig.2003.07.003.
The role of p38 mitogen-activated protein (MAP) kinase in the activation of human neutrophils and repression of TNF-alpha-induced apoptosis in response to plasma opsonized crystals of calcium pyrophosphate dihydrate (CPPD) was investigated. We monitored the endogenous phosphotransferase activity of p38 kinase in neutrophils stimulated with CPPD crystals (25 mg/ml) alone or in the presence of TNF-alpha (10 ng/ml), and with TNF-alpha alone. CPPD crystals induced a 2-fold activation of p38 kinase activity over the basal activity that was observed in untreated neutrophils. Furthermore, CPPD crystals repressed the TNF-alpha associated 6-fold induction of p38 kinase phosphotransferase activity to levels associated with CPPD crystal incubation alone in a PD98059 (20 ng/ml) and Wortmannin (100 nM) sensitive manner. Inhibition of CPPD crystal-induced activation of the neutrophil inflammatory response as measured by chemiluminescence, superoxide anion generation and degranulation as determined by myeloperoxidase and lysozyme release was observed in the presence of the specific p38 MAP kinase inhibitor SB203580 (5 microM). CPPD crystal associated repression of TNF-alpha-induced activation of neutrophil apoptosis as determined by DNA fragmentation correlated with the CPPD crystal mediated inhibition of p38 kinase activity, probably through crystal inhibition of caspase 3. Together, our results indicate that the CPPD crystal associated inflammatory response is regulated through the activation of p38 kinase to sub-apoptotic levels, and that the repression of the TNF-alpha-induced apoptosis program in neutrophils is mediated via the repression of caspase 3 mediated apoptosis-associated p38 kinase activity.
研究了p38丝裂原活化蛋白（MAP）激酶在人类中性粒细胞活化以及对血浆调理的二水焦磷酸钙（CPPD）晶体反应中抑制肿瘤坏死因子-α（TNF-α）诱导的细胞凋亡中的作用。我们监测了单独用CPPD晶体（25 mg/ml）刺激或在TNF-α（10 ng/ml）存在下刺激的中性粒细胞中p38激酶的内源性磷酸转移酶活性，以及单独用TNF-α刺激的中性粒细胞中p38激酶的内源性磷酸转移酶活性。与未处理的中性粒细胞中观察到的基础活性相比，CPPD晶体诱导p38激酶活性增加了2倍。此外，CPPD晶体以对PD98059（20 ng/ml）和渥曼青霉素（100 nM）敏感的方式，将TNF-α相关的p38激酶磷酸转移酶活性6倍诱导抑制至仅与CPPD晶体孵育相关的水平。在存在特异性p38 MAP激酶抑制剂SB203580（5 microM）的情况下，观察到通过化学发光测量的CPPD晶体诱导的中性粒细胞炎症反应活化受到抑制，通过髓过氧化物酶和溶菌酶释放测定的超氧阴离子生成和脱颗粒也受到抑制。通过DNA片段化测定，CPPD晶体相关的对TNF-α诱导的中性粒细胞凋亡活化的抑制与CPPD晶体介导的p38激酶活性抑制相关，可能是通过晶体对caspase 3的抑制。总之，我们的结果表明，CPPD晶体相关的炎症反应通过将p38激酶激活至亚凋亡水平来调节，并且中性粒细胞中TNF-α诱导的凋亡程序的抑制是通过抑制caspase 3介导的凋亡相关p38激酶活性来介导的。
