The caspase inhibitor IDN-6556 attenuates hepatic injury and fibrosis in the bile duct ligated mouse
The caspase inhibitor IDN-6556 attenuates hepatic injury and fibrosis in the bile duct ligated mouse
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DOI：
10.1124/jpet.103.060129
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发表时间：
2004-03-01
影响因子：
3.5
通讯作者：
Gores, GJ
中科院分区：
文献类型：
--
作者：
Canbay, A;Feldstein, A;Gores, GJ
肝损伤的特征是肝细胞凋亡和产生胶原蛋白的肝星状细胞（HSC）活化。肝细胞凋亡促进肝损伤和纤维化，而激活的HSC凋亡则限制肝纤维化。肝细胞凋亡的药理抑制可能通过阻止肝细胞凋亡来减轻肝损伤和纤维化，或通过允许活化的 HSC 积聚来促进纤维化。为了确定抑制肝细胞凋亡对肝损伤、炎症和肝纤维形成的净效应，我们检查了胰蛋白酶抑制剂 IDN-6556 对胆管结扎 (BDL) 小鼠这些参数的影响。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记测定和活性半胱天冬酶 3/7 的免疫荧光来评估肝细胞凋亡，并通过组织病理学和血清丙氨酸氨基转移酶 (ALT) 测定来评估肝损伤。使用实时聚合酶链反应来测量肝脏炎症、HSC 激活和纤维化标志物的 mRNA 转录本。对 α-平滑肌肌动蛋白进行免疫组织化学分析以鉴定 HSC 激活。通过天狼星红染色和数字成像技术对胶原沉积进行定量。与生理盐水治疗的 3 天 BDL 小鼠相比，IDN-6556 治疗的肝细胞凋亡和肝损伤（胆汁梗塞和血清 ALT 值）减少。肝脏炎症标志物 [趋化因子 (C-X-C) 配体 1 和巨噬细胞炎症蛋白 2 趋化因子表达] 和肝纤维发生（转化生长因子-β 和胶原蛋白 I 表达）也减弱。与这些数据一致，通过 α-平滑肌肌动蛋白 mRNA 表达和免疫组织化学评估，在 3 天和 10 天 BDL 动物中，HSC 活化均显着降低。总的来说，这些数据表明肝细胞凋亡引发级联反应，最终导致肝损伤和纤维化。泛半胱天冬酶抑制剂 IDN-6556 是一种有前途的治疗胆汁淤积性肝损伤的药物。
Liver injury is characterized by hepatocyte apoptosis and collagen-producing activated hepatic stellate cells ( HSC). Hepatocyte apoptosis promotes liver injury and fibrosis, whereas activated HSC apoptosis limits hepatic fibrosis. Pharmacological inhibition of liver cell apoptosis may potentially attenuate liver injury and fibrosis by blocking hepatocyte apoptosis or promote fibrosis by permitting accumulation of activated HSCs. To ascertain the net effect of inhibiting liver cell apoptosis on liver injury, inflammation, and hepatic fibrogenesis, we examined the effect of a pancaspase inhibition IDN-6556 on these parameters in the bile duct ligated (BDL) mouse. Hepatocyte apoptosis was assessed by the terminal deoxynucleotidyl transferase dUTP nick-end labeling assay and immunofluorescence for active caspases 3/7, and liver injury by histopathology and serum alanine aminotransferase (ALT) determinations. Real-time polymerase chain reaction was used to measure mRNA transcripts for markers of hepatic inflammation, HSC activation, and fibrosis. Immunohistochemistry for alpha-smooth muscle actin was performed to identify HSC activation. Collagen deposition was quantitated by Sirius red staining and digital imaging techniques. Hepatocyte apoptosis and liver injury (bile infarcts and serum ALT values) were reduced in IDN-6556-treated versus saline-treated 3-day BDL mice. Markers for liver inflammation [ chemokine (C-X-C) ligand 1 and macrophage inflammatory protein-2 chemokine expression] and hepatic fibrogenesis ( transforming growth factor-beta and collagen I expression) were also attenuated. Consistent with these data, HSC activation as assessed by alpha-smooth muscle actin mRNA expression and immunohistochemistry was markedly reduced in both 3- and 10-day BDL animals. Collectively, these data suggest hepatocyte apoptosis initiates cascades culminating in liver injury and fibrosis. The pan-caspase inhibitor IDN-6556 is a promising agent for cholestatic liver injury.