State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, Sichuan Province, China.
World J Gastroenterol. 2012 Feb 21;18(7):654-61. doi: 10.3748/wjg.v18.i7.654.
To evaluate the hepatoprotective roles of (Z)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione (SKLB010) against carbon tetrachloride (CCl₄)-induced acute and chronic liver injury and its underlying mechanisms of action.
In the first experiment, rats were weighed and randomly divided into 5 groups (five rats in each group) to assess the protective effect of SKLB010 on acute liver injury. For induction of acute injury, rats were administered a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl₄ dissolved in olive oil (1:1). Group 1 was untreated and served as the control group; group 2 received CCl₄ for induction of liver injury and served as the model group. In groups 3, 4 and 5, rats receiving CCl₄ were also treated with SKLB010 at doses of 25, 50 and 100 mg/kg, respectively. Blood samples were collected at 6, 12 and 24 h after CCl₄ intoxication to determine the serum activity of alanine amino transferase. Tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β) were determined using enzyme-linked immunosorbent assay. At 24 h after CCl₄ injection, liver fibrogenesis was evaluated by hematoxylin-eosin (HE) staining and immunohistochemical analyses. Cytokine transcript levels of TNF-α, IL-1β and inducible nitric oxide synthase in the liver tissues of rats were measured using a reverse transcriptase reverse transcription-polymerase chain reaction technique. In the second experiment, rats were randomly divided into 2 groups (15 rats in each group), and liver injury in the CCl₄-administered groups was induced by a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl₄ dissolved in olive oil (1:1). The SKLB010-treated groups received oral 100 mg/kg SKLB010 before CCl₄ administration. Five rats in each group were sacrificed at 2 h, 6 h, 12 h after CCl₄ intoxication and small fortions of livers were rapidly frozen for extraction of total RNA, hepatic proteins and glutathione (GSH) assays. In the hepatic fibrosis model group, rats were randomly divided into 2 groups (5 rats each group). Rats were injected intraperitoneally with a mixture of CCl₄ (1 mL/kg body weight) and olive oil [1:1 (v/v)] twice a week for 4 wk. In the SKLB010-treated groups, SKLB010 (100 mg/kg) was given once daily by oral gavage for 4 wk after CCl₄ administration. The rats were sacrificed one week after the last injection and the livers from each group were harvested and fixed in 10% formalin for HE and immunohistochemical staining.
In this rat acute liver injury model, oral administration of SKLB010 blocked liver tissue injury by down-regulating the serum levels of alanine aminotransferase, suppressing inflammatory infiltration to liver tissue, and improving the histological architecture of liver. SKLB010 inhibited the activation of NF-κB by suppressing the degradation of IκB, and prevented the secretion of pro-inflammatory mediators such as tumor necrosis factor-α, interleukin-1β, and the reactive free radical, nitric oxide, at the transcriptional and translational levels. In this chronic liver fibrosis model, treatment with 100 mg/kg per day SKLB010 attenuated the degree of hepatic fibrosis and area of collagen, and blocked the accumulation of smooth-muscle actin-expressed cells.
These results suggest that SKLB010 is a potent therapeutic agent for the treatment of CCl₄-induced hepatic injury.
评估（Z）-5-（4-甲氧基苄叉）噻唑烷-2,4-二酮（SKLB010）对四氯化碳（CCl4）诱导的急性和慢性肝损伤的保护作用及其作用机制。
在第一个实验中，大鼠称重后随机分为 5 组（每组 5 只大鼠），评估 SKLB010 对急性肝损伤的保护作用。为诱导急性损伤，大鼠腹腔内注射 2 毫升/千克 50%（v/v）的 CCl4 溶于橄榄油（1:1）。第 1 组未处理，作为对照组；第 2 组给予 CCl4 诱导肝损伤，作为模型组。第 3、4 和 5 组大鼠给予 CCl4 后，分别给予 25、50 和 100 mg/kg 的 SKLB010 治疗。CCl4 中毒后 6、12 和 24 小时采集血样，测定血清丙氨酸氨基转移酶活性。采用酶联免疫吸附试验测定肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）。CCl4 注射后 24 小时，通过苏木精-伊红（HE）染色和免疫组织化学分析评估肝纤维化。采用逆转录-聚合酶链反应技术测定大鼠肝组织中 TNF-α、IL-1β 和诱导型一氧化氮合酶的细胞因子转录水平。在第二个实验中，大鼠随机分为 2 组（每组 15 只大鼠），CCl4 给药组通过腹腔注射 2 毫升/千克 50%（v/v）的 CCl4 溶于橄榄油（1:1）诱导肝损伤。SKLB010 治疗组在 CCl4 给药前给予 100 mg/kg SKLB010 口服。CCl4 中毒后 2、6、12 小时，每组各取 5 只大鼠处死，迅速将肝脏的小部分冷冻用于提取总 RNA、肝蛋白和谷胱甘肽（GSH）测定。在肝纤维化模型组中，大鼠随机分为 2 组（每组 5 只大鼠）。大鼠每周两次腹腔注射 CCl4（1 毫升/千克体重）和橄榄油[1:1（v/v）]，共 4 周。在 SKLB010 治疗组中，在 CCl4 给药后，每日口服给予 100 mg/kg 的 SKLB010 治疗 4 周。最后一次注射后一周处死大鼠，收集各组肝脏，用 10%福尔马林固定进行 HE 和免疫组织化学染色。
在该大鼠急性肝损伤模型中，口服 SKLB010 通过下调血清丙氨酸氨基转移酶水平、抑制炎性浸润到肝组织和改善肝组织学结构来阻止肝组织损伤。SKLB010 通过抑制 IκB 的降解来抑制 NF-κB 的激活，并防止肿瘤坏死因子-α、白细胞介素-1β 和活性自由基（一氧化氮）等促炎介质在转录和翻译水平的分泌。在该慢性肝纤维化模型中，每天给予 100 mg/kg SKLB010 可减轻肝纤维化程度和胶原面积，并阻止平滑肌肌动蛋白表达细胞的积聚。
这些结果表明，SKLB010 是治疗 CCl4 诱导的肝损伤的有效治疗药物。
