Calcium Contributes to the Cytotoxic Interaction Between Diclofenac and Cytokines.
Calcium Contributes to the Cytotoxic Interaction Between Diclofenac and Cytokines.
复制标题
钙有助于双氯芬酸和细胞因子之间的细胞毒性相互作用。
DOI：
10.1093/toxsci/kfv249
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Roth,RobertA
中科院分区：
文献类型：
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作者：
Maiuri,AshleyR;Breier,AnnaB;Turkus,JonathanD;Ganey,PatriciaE;Roth,RobertA
双氯芬酸（DCLF）是一种广泛使用的非甾体抗炎药，与人类特异质药物性肝损伤（IDILI）相关。DCLF诱导的肝损伤的机制尚不清楚;然而，某些炎性疾病患者发生IDILI的风险增加，这增加了免疫介质在发病机制中发挥作用的可能性。DCLF与细胞因子肿瘤坏死因子-α（TNF）和干扰素-γ（IFN）协同作用，通过激活内质网（ER）应激反应途径和丝裂原活化蛋白激酶、c-Jun N-末端激酶（JNK）和细胞外信号调节激酶（ERK）的机制在体外引起肝细胞凋亡。DCLF还导致肝细胞内钙（Ca++）增加，但其在DCLF和细胞因子之间的细胞毒性协同作用中的作用尚不清楚。我们测试了Ca++有助于DCLF/苦参碱诱导的细胞毒性协同作用的假设。用DCLF处理HepG 2细胞导致细胞内Ca++在6和12 h增加，并且在TNF和IFN存在下在12 h增强这种反应。细胞内Ca++螯合剂BAPTA/AM降低DCLF/细胞因子共处理引起的细胞毒性和caspase-3激活。BAPTA/AM还显著降低了DCLF诱导的ER应激传感器、蛋白激酶RNA样ER激酶（PERK）的活化以及JNK和ERK的活化。用三磷酸肌醇受体拮抗剂处理细胞几乎完全消除了DCLF/苦参碱诱导的细胞毒性，并降低了DCLF诱导的PERK、JNK和ERK活化。这些发现表明，Ca++通过促进ER应激反应途径和JNK和ERK的活化而有助于DCLF/苦参碱诱导的细胞毒性协同作用。
Diclofenac (DCLF) is a widely used non-steroidal anti-inflammatory drug that is associated with idiosyncratic, drug-induced liver injury (IDILI) in humans. The mechanisms of DCLF-induced liver injury are unknown; however, patients with certain inflammatory diseases have an increased risk of developing IDILI, which raises the possibility that immune mediators play a role in the pathogenesis. DCLF synergizes with the cytokines tumor necrosis factor-alpha (TNF) and interferon-gamma (IFN) to cause hepatocellular apoptosisin vitroby a mechanism that involves activation of the endoplasmic reticulum (ER) stress response pathway and of the mitogen-activated protein kinases, c-Jun N-terminal kinase (JNK), and extracellular signal-regulated kinase (ERK). DCLF also causes an increase in intracellular calcium (Ca++) in hepatocytes, but the role of this in the cytotoxic synergy between DCLF and cytokines is unknown. We tested the hypothesis that Ca++contributes to DCLF/cytokine-induced cytotoxic synergy. Treatment of HepG2 cells with DCLF led to an increase in intracellular Ca++at 6 and 12 h, and this response was augmented in the presence of TNF and IFN at 12 h. The intracellular Ca++chelator BAPTA/AM reduced cytotoxicity and caspase-3 activation caused by DCLF/cytokine cotreatment. BAPTA/AM also significantly reduced DCLF-induced activation of the ER stress sensor, protein kinase RNA-like ER kinase (PERK), as well as activation of JNK and ERK. Treatment of cells with an inositol trisphosphate receptor antagonist almost completely eliminated DCLF/cytokine-induced cytotoxicity and decreased DCLF-induced activation of PERK, JNK, and ERK. These findings indicate that Ca++contributes to DCLF/cytokine-induced cytotoxic synergy by promoting activation of the ER stress-response pathway and JNK and ERK.