Pemphigus serum and captopril induce heat shock protein 70 and inducible nitric oxide synthase overexpression, triggering apoptosis in human keratinocytes
Pemphigus serum and captopril induce heat shock protein 70 and inducible nitric oxide synthase overexpression, triggering apoptosis in human keratinocytes
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DOI：
10.1111/j.1365-2133.2004.05919.x
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发表时间：
2004-06-01
影响因子：
10.3
通讯作者：
Ruocco, V
中科院分区：
文献类型：
--
作者：
Baroni, A;Buommino, E;Ruocco, V
背景：卡托普利是一种化学结构中含有巯基的血管紧张素转换酶抑制剂。它通常被用作一种降压药。在接受卡托普利治疗的患者中，寻常型天疱疮的发生已被反复报道。卡托普利和天疱疮血清在体内或体外诱导棘层松解的能力已被实验证明。目的探讨卡托普利和天疱疮血清分别通过生物化学和免疫学机制诱导角质形成细胞发生凋亡的机制。提取DNA，用末端脱氧核苷酸转移酶介导的脱氧尿嘧啶核苷缺口末端标记法检测细胞凋亡。聚合酶链式反应(PCR)结果显示，与未处理组相比，处理组细胞中P53、c-myc和诱导型一氧化氮合酶(INOS)基因表达增加。诱导型一氧化氮合酶基因表达的增加与一氧化氮的过量产生有关。此外，在用卡托普利或天疱疮血清处理的细胞中，加入1 mmoL的L-1 N-单甲基-L-精氨酸(一种精氨酸的结构类似物)，亚硝酸盐水平降低了约70%。Western印迹分析显示，在卡托普利或天疱疮血清处理的细胞中热休克蛋白70(HSP70)过表达。结论卡托普利或天疱疮血清可诱导角质形成细胞发生凋亡，诱导型一氧化氮合酶基因和热休克蛋白70参与细胞程序性死亡的级联反应。
Background Captopril is an angiotensin-converting enzyme inhibitor with sulphydryl groups in its chemical structure. It is commonly used as an antihypertensive drug. The occurrence of pemphigus vulgaris has repeatedly been reported in patients receiving captopril. The capacity of captopril and pemphigus serum to induce acantholysis, in vivo or in vitro, has been demonstrated experimentally. Objectives To show that captopril and pemphigus serum, acting by a biochemical and immunological mechanism, respectively, trigger apoptosis.Methods Human keratinocyte cells were treated with 15 mmol L-1 captopril or with pemphigus serum. DNA was extracted and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling method was used to detect apoptosis.Results DNA fragmentation occurred after 72 h of treatment. Increased expression of p53, c-myc and inducible nitric oxide (NO) synthase (iNOS) mRNA were observed by polymerase chain reaction (PCR) in the treated cells compared with the untreated ones. The increase in iNOS gene expression was associated with overproduction of NO. Moreover, the addition of 1 mmol L-1 N-monomethyl-L-arginine, a structural analogue of arginine, reduced nitrite levels by about 70% in cells treated with captopril or pemphigus serum. Western blot analysis revealed an overexpression of heat shock protein 70 (hsp70) in cells treated with captopril or pemphigus serum. Finally, total inhibition of the keratinocyte transglutaminase gene was shown by PCR analysis in the same samples, compared with control cells.Conclusions These data demonstrate the involvement of apoptosis in keratinocytes treated with captopril or pemphigus serum, with induction of the iNOS gene and hsp70 in the cascade of events leading to programmed cell death.