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目的探讨非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)大鼠肝脏硫化氢(hydrogen sulfide,H2S)变化及与肝细胞线粒体损伤的可能关系。方法采用雄性SD大鼠26只,分为正常对照组(n=6),NASH模型组(n=10)及复方牛胎肝提取物治疗组(n=10),高脂饮食12周建立NASH大鼠模型。亚甲基蓝分光光度法测定肝组织H2S生成率,RT-PCR及免疫组织化学测定肝组织CSEmRNA及其表达。透射电子显微镜观察肝细胞超微结构、PCR法测定肝细胞线粒体DNA(mtDNA)片段ND1、ND6、CO1、CYB及ATP6。结果 NASH大鼠模型组肝细胞线粒体肿大、膜模糊或嵴消失,正常组大鼠mtDNA片段ND1、ND6、CO1、CYB及ATP6表达明显高于模型组和治疗组(P<0.05)。NASH模型组肝组织H2S生成率〔(1.26±0.08)nmol/min〕较正常组〔(2.11±0.17)nmol/min〕明显减少(P<0.05),且肝脏CSEmRNA水平及其表达减少,治疗组与模型组比较差异无统计学意义。结论 NASH大鼠肝脏H2S合成减少可能与肝细胞线粒体损伤有关,H2S在NASH发病机制中可能具有保护肝脏的作用。
Objective To investigate the changes of hepatic hydrogen sulfide (H2S) and its possible relationship with hepatocyte mitochondrial damage in nonalcoholic steatohepatitis (NASH) rats. Methods Twenty-six male Sprague-Dawley rats were randomly divided into normal control group (n = 6), NASH model group (n = 10) and compound bovine fetal liver extract treatment group (n = 10) Rat model. H2S production in liver tissue was determined by methylene blue spectrophotometry, and expression of CSE mRNA in liver tissue was detected by RT-PCR and immunohistochemistry. The ultrastructure of hepatocytes was observed by transmission electron microscopy. The mitochondrial DNA (mtDNA) fragments of ND1, ND6, CO1, CYB and ATP6 were detected by PCR. Results The mitochondria of hepatocytes in the NASH model group were swollen and the membrane was fuzzy or disappeared. The expression of mtDNA ND1, ND6, CO1, CYB and ATP6 in normal group were significantly higher than those in model group and treatment group (P <0.05). Compared with the normal group (2.11 ± 0.17) nmol / min〕 〔(1.26 ± 0.08) nmol / min〕 in NASH model group significantly reduced the level of liver CSE mRNA expression and its expression, the treatment group Compared with the model group, the difference was not statistically significant. Conclusions The reduction of H2S synthesis in the liver of NASH rats may be related to the mitochondrial damage of liver cells. H2S may play a role in liver protection in the pathogenesis of NASH.