Fiorucci S, Antonelli E, Santucci L, Morelli O, Miglietti M, Federici B, Mannucci R, Del Soldato P, Morelli A
Sezione di Gastroenterologia ed Epatologia, Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi di Perugia, Perugia, Italy.
Gastroenterology. 1999 May;116(5):1089-106. doi: 10.1016/s0016-5085(99)70012-0.
BACKGROUND & AIMS: Caspases, a class of cysteine proteases, modulate apoptosis. Nitric oxide (NO)-releasing nonsteroidal anti-inflammatory drugs (NSAIDs) are a new class of NSAID derivatives with reduced gastrointestinal toxicity. The aim of this study was to investigate whether cysteine endoproteases are involved in the pathogenesis of NSAID gastropathy and are target for NO-aspirin (NCX-4016).
Rats were treated orally with aspirin or equimolar doses of NCX-4016. Caspase activities were measured by fluorometric assay. Apoptosis was quantified by an enzyme-linked immunosorbent assay for histone-associated DNA, DNA ladder on agarose gel, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay. A primary culture of gastric chief cells was used to investigate whether NCX-4016 modulates guanosine 3',5'-cyclic monophosphate (cGMP)-dependent pathways.
Short- and long-term (7 days) aspirin administration resulted in a time- and dose-dependent gastric injury that was associated with apoptosis and caspase up-regulation. Z-VAD.FMK, a pancaspase inhibitor, and NO donors protected from acute damage induced by aspirin. NCX-4016 spared the gastric mucosa and caused caspase inactivation by S-nitrosylation. Inhibition of tumor necrosis factor (TNF)-alpha release or activity by TAPI-2 or anti-TNF-alpha receptor monoclonal antibodies protected against mucosal damage and caspase activation. NCX-4016 protected gastric chief cells from toxicity induced by TNF-alpha by activating cGMP-dependent pathways.
Aspirin administration leads to a TNF-alpha-dependent activation of gastric caspases. NO-aspirin spares the gastric mucosa and inhibits caspase activity through cGMP-dependent and -independent pathways.
半胱天冬酶是一类半胱氨酸蛋白酶，可调节细胞凋亡。释放一氧化氮（NO）的非甾体抗炎药（NSAIDs）是一类新的NSAID衍生物，其胃肠道毒性降低。本研究的目的是调查半胱氨酸内切蛋白酶是否参与NSAID胃病的发病机制，以及是否是NO-阿司匹林（NCX-4016）的作用靶点。
给大鼠口服阿司匹林或等摩尔剂量的NCX-4016。通过荧光测定法测量半胱天冬酶活性。通过用于组蛋白相关DNA的酶联免疫吸附测定、琼脂糖凝胶上的DNA梯带以及末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记测定来定量细胞凋亡。使用胃主细胞的原代培养物来研究NCX-4016是否调节鸟苷3',5'-环磷酸（cGMP）依赖性途径。
短期和长期（7天）给予阿司匹林导致时间和剂量依赖性的胃损伤，这与细胞凋亡和半胱天冬酶上调有关。泛半胱天冬酶抑制剂Z-VAD.FMK和NO供体可保护免受阿司匹林诱导的急性损伤。NCX-4016可保护胃黏膜，并通过S-亚硝基化导致半胱天冬酶失活。TAPI-2或抗TNF-α受体单克隆抗体抑制肿瘤坏死因子（TNF）-α释放或活性可保护免受黏膜损伤和半胱天冬酶激活。NCX-4016通过激活cGMP依赖性途径保护胃主细胞免受TNF-α诱导的毒性。
给予阿司匹林会导致胃半胱天冬酶的TNF-α依赖性激活。NO-阿司匹林可保护胃黏膜，并通过cGMP依赖性和非依赖性途径抑制半胱天冬酶活性。
