组蛋白去乙酰化酶抑制剂ACY1215对急性肝衰竭大鼠肝细胞线粒体的保护作用*

doi:10.3969/j.issn.1672-5069.2017.06.009

摘要/Abstract

摘要： 目的探讨ACY1215对急性肝衰竭大鼠肝细胞线粒体损伤的保护作用及其相关机制。方法24只SD大鼠被随机分为对照组、模型组和ACY1215处理组。以脂多糖（LPS）联合D-氨基半乳糖（D-Gal）注射建立急性肝衰竭大鼠模型,药物干预组在建立急性肝衰竭组前2 h给予ACY1215（10 mg·kg-1）注射。造模48 h后处死动物,在光镜及电镜下观察肝组织学变化,采用梯度离心法分离肝细胞线粒体,采用荧光酶标法测定线粒体膜通透性转换孔（MPTP）,采用ELISA法测定线粒体细胞色素C（Cytc）含量,常规测定血清ALT、AST和总胆红素（TBil）水平。结果与模型组比,ACY1215处理组肝组织学破坏程度减轻,电镜显示肝细胞线粒体肿胀减轻;急性肝衰竭模型组大鼠血清ALT、AST和TBIL水平分别为（5114.1±252.6） U/L、（2909.8±31.7） U/L和（97.6±1.4）μmol/L,均显著高于正常组大鼠的（56.0±4.4） U/L、（130.4±12.6） U/L和（7.4±0.7）μmol （P均<0.05）,但处理组ALT、AST和TBIL水平均较模型组减低,分别为（799.4±11.2） U/L、（401.0±5.6） U/L和（28.0±1.2） μmol /L（P均<0.05）;急性肝衰竭模型组大鼠MPTP相对荧光值（RFU）为（96822.0±16733.1） RFU/mgprot,较正常组大鼠的（156300.0±24043.3）RFU/mgprot显著升高（P<0.05）,也显著高于处理组的（127150.0±12337.6） RFU/mgprot（P<0.05）;模型组大鼠线粒体内Cytc为（0.17±0.03） ng/mgprot,较正常组大鼠线粒体内的（0.39±0.10） ng/mgprot显著减少（P<0.05）,也低于处理组的（0.32±0.06） ng/mgprot （P<0.05）。结论ACY1215对急性肝衰竭大鼠肝脏线粒体有一定的保护作用,其作用机制可能是通过抑制肝细胞线粒体MPTP的开放,从而减少了Cytc进入到细胞质内有关。

关键词: 急性肝衰竭, 线粒体, 组蛋白去乙酰化抑制剂 ACY1215, 大鼠

Abstract: Objective To investigate the protective mechanism of histone deacetylase inhibitor ACY1215 on liver mitochondrial damage in rats with acute hepatic failure（AHF）. Methods 24 male SD rats were randomly divided into control group,model group and ACY1215-intervened group. Lipopolysaccharide（LPS） and D-galactosamine （D-Gal） were used to establish acute liver failure model in rats. The ACY1215 （10 mg·kg-1） was given 2 hours before the establishment of acute liver failure. Serum and liver samples of rats were obtained at end of 48 hours after AHF induction. The liver histological changes were observed by light and electron microscopy,and the mitochondria of hepatocytes were separated by gradient centrifugation. The relative fluorescence （RFU） of mitochondrial membrane permeability transition porosity （MPTP） was tested by fluorescence spectrometry. The content of cytochrome C（Cytc） in mitochondria supernatants was determined by ELISA. The contents of serum ALT,AST and total bilirubin were detected by routine biochemical method. Results Compared with in the model group,the degree of liver histological damage was reduced in the ACY1215-intervened group,and the mitochondrial swelling of the hepatocytes was improved;serum ALT [（5114.1±252.6） U/L vs. （56.0±4.4） U/L],AST [（2909.8±31.7） U/L vs. （130.4±12.6） U/L] and total bilirubin levels [（97.6±1.4） μmol/L vs. （7.4±0.7） μmol/L] in the model group were significantly higher than those in the control group （P<0.05）,while they[（799.4±11.2） U/L,（401.0±5.6） U/L and（28.0±1.2） μmol/L,respectively] in the ACY1215-intervened group significantly decreases as compared to those in the model group（P<0.05）;the MPTP in acute liver failure model group was significantly higher than that in the normal control group[（968220.0±16733.1） RFU/mgprot vs.（156300.0±24043.3）RFU/mgprot，P<0.05）]，and also higher than in the ACY1215-intervened group [（127150.0±12337.6） RFU/mgprot，P<0.05]；the mitochondrial Cytc level in the model group was significantly lower than that in the control group [（0.17±0.03） ng/mgprot vs. （0.39±0.10） ng/mgprot，P<0.05]，and also lower than that in the ACY1215-intervened group [（0.32±0.06） ng/mgprot，P<0.05]. Conclusion ACY1215 has a protective effect on liver mitochondria in rats with acute hepatic failure，and the mechanism might be the inhibition of opening of mitochondrial MPTP in the livers and reduction of translocation of Cytc into the cytoplasm.

Key words: Acute hepatic failure, Mitochondria, Histone deacetylase inhibitor ACY1215, Rats

陈倩, 焦方舟, 张海月, 张文斌, 刘扬, 龚作炯. 组蛋白去乙酰化酶抑制剂ACY1215对急性肝衰竭大鼠肝细胞线粒体的保护作用*[J]. 实用肝脏病杂志, 2017, 20(6): 672-675.

Chen Qian, Jiao Fangzhou, Zhang Haiyue. Protective effect of histone deacetylase inhibitor ACY1215 on hepatocyte mitochondria in rats with acute hepatic failure[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2017, 20(6): 672-675.

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