Molecular mechanism of inhaled anesthetic-induced hepatotoxicity

吸入麻醉药肝毒性的分子机制

• 批准号: 11671499
• 负责人: HIRAKAWA Masahisa
• 金额: $ 2.43万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671499/
• 关键词: inhaled anesthetics; hepatotoxicity; heme; heme oxygenase-1; cytochrome P450-2E1; oxidative stress; CYP2E1遺伝子導入肝細胞; ヘム; 吸収麻酔薬; CYP2E1; 四塩化炭素; CYP2E1遺伝子導入細胞; イソフルラン; ハロタン; ヘムオキシゲナーゼ-1; ストレス蛋白; 低酸素; ハロタン肝障害; ヘムオキシゲナーゼ

We investigated the molecular mechanism of inhaled anesthetic-induced hepatotoxicity in vivo and in vitro. Exposure of phenobarbital-pretreated rats to halothane-hypoxia caused a rapid increase in cytosolic free heme concentration, which was preceded by a significant decrease in microsomal cytochrome P450 (CYP) content in the liver. There were also marked induction in heme oxygenase-1 (HO-1). hemin pretreatment of these animals not only induced hepatic HO-1 , but also almost completely abrogated the halothane-induced hepatotoxicity. These findings indicate that halothane-induced hepatotoxicity is due to an increase in hepatic free heme concentration that is a potent prooxidant, and HO-1 induction is an important protective response against such changes. Next, we examined the induction of HSP70 and HO-1, in rat livers pretreated with phenobarbital, followed by exposure to isoflurane, or halothane under hypoxic condition. The induction of HSP70 was observed by halothane-hypoxia treatment … More is higher than that by isoflurane-hypoxia treatment. Serum alanine aminotransferase (ALT) activity correlated well with the extent of centrilobular necrosis, showed similar changes with increases in HSP70. In contrast, HO-1 was induced only by halothane-hypoxia treatment, but not by other treatments. These findings demonstrate that there is a significant difference in hepatic injury, HO-1 and HSP70 induction, between halothane-hypoxia and isoflurane-hypoxia. Isoflurane is known to be safer than halothane, which may be in part accounted for by its lesser oxidative stress as assessed by a smaller induction of HSPs than halothane treatment. Finally, since inhaled anesthetics are metabolized by cytochrome P450-2E1 (CYP2E1) and inhaled anesthetics are derivates of carbon tetrachloride, the cytotoxic effects of CCl_4 in a liver cell line expressing CYP2E1 (HLE/2E1) in comparison to those in the mother cell line (HLE) were determined. The effects of CCl_4 on the gene expression of HSP70, a potential marker of oxidative stress, were also examined. The viability of HLE/2E1 cells after exposure to CCl_4 was significantly decreased compared with that of HLE cells. Northern blot analysis revealed that the HSP70 mRNA level was significantly increased after CCl_4 treatment in both cell lines, while the magnitude of its increase was much greater in HLE/2E1 cells than HLE cells. These results suggest that the oxidative stress induced by CYP2E1 plays an important role in the increase in cytotoxicity of CCl_4 in CYP2E1-overexpressing cells and that CYP2E1-overexpressing human liver cell line may be suitable for investigating the hepatotoxicity of volatile anesthetics. Less

我们在体内和体外研究了吸入麻醉药引起肝毒性的分子机制。暴露苯巴比妥预处理大鼠氟烷缺氧引起细胞溶质游离血红素浓度的迅速增加，这是由肝微粒体细胞色素P450（P450）含量的显着下降之前。血红素氧合酶-1（HO-1）也有明显的诱导作用。氯化血红素预处理这些动物不仅诱导肝HO-1，但也几乎完全消除氟烷诱导的肝毒性。这些研究结果表明，氟烷诱导的肝毒性是由于肝游离血红素浓度的增加，这是一种有效的促氧化剂，HO-1诱导是一个重要的保护性反应，对这种变化。接下来，我们研究了诱导HSP 70和HO-1，在大鼠肝脏预处理苯巴比妥，然后暴露于异氟烷，或氟烷在缺氧条件下。氟烷低氧诱导HSP 70表达 ...更多信息 高于异氟醚低氧处理。血清丙氨酸氨基转移酶（ALT）活性与小叶中心坏死的程度相关，与HSP 70的增加表现出类似的变化。而HO-1仅在氟烷-低氧处理下被诱导，其他处理均不诱导。这些结果表明，氟烷缺氧和异氟烷缺氧对肝损伤、HO-1和HSP 70的诱导有显著差异。已知异氟烷比氟烷更安全，这可能部分是由于其氧化应激更低，如通过比氟烷治疗更小的HSP诱导所评估的。最后，由于吸入麻醉药是由细胞色素P450- 2 E1（CYP 2 E1）代谢的，而吸入麻醉药是四氯化碳的衍生物，因此测定了CCl_4对表达CYP 2 E1的肝细胞系（HLE/2 E1）和母细胞系（HLE）的细胞毒性作用。此外，还检测了CCl_4对氧化应激的潜在标志物HSP 70基因表达的影响。HLE/2 E1细胞经CCl_4处理后，其存活率明显低于HLE细胞。北方印迹分析显示，CCl_4处理后，HLE/2 E1和HLE/2 E1细胞中HSP 70 mRNA的表达均显著增加，但HLE/2 E1细胞中HSP 70 mRNA的表达量明显高于HLE细胞。提示CYP 2 E1诱导的氧化应激在CCl_4对CYP 2 E1过表达细胞的毒性增强中起重要作用，CYP 2 E1过表达细胞系可用于研究挥发性麻醉药的肝毒性。少

项目成果

Yasuo Odaka, et.al. 9persons: "Prevention of halothane-induced hepatotoxicity by hemin pretreatment: The protective role of heme oxygenase-1 induction"Biochemical Pharmacology. Vol.59, No.6(印刷中). (2000)

Yasuo Odaka，等9人：“通过血红素预处理预防氟烷诱导的肝毒性：血红素加氧酶-1诱导的保护作用”《生物化学药理学》第59卷，第6期（出版中）。

Akira Yamasaki, et al., 6 persons: "Differential effects of isoflurane and halothane of the induction of heat shock proteins"Biochemical Pharmacology. Vol.62. 375-382 (2001)

Akira Yamasaki等，6人：“异氟烷和氟烷对热休克蛋白诱导的不同作用”生化药理学。

Shuji Takahashi, et al., 8 persons: "Increased cytotoxicity of carbon tetrachloride in a human hepatoma cell line (HLE/2E1) overexpressing cytochrome P450 2E1"The Journal of International Medical Research. Vol.30(印刷中). (2002)

Shuji Takahashi 等，8 人：“四氯化碳在过表达细胞色素 P450 2E1 的人肝癌细胞系 (HLE/2E1) 中的细胞毒性增加”《国际医学研究杂志》第 30 卷（出版中）。

Akira Yamasaki, et al.,6 persons: "Differential effects of isoflurane and halothane on the induction of heat shock proteins"Biochemical Pharmacology. (印刷中). (2001)

Akira Yamasaki 等人，6 人：“异氟烷和氟烷对热休克蛋白诱导的不同影响”生化药理学（2001 年出版）。