褪黑素对肝内胆汁淤积大鼠肝组织NO、MDA、GSH及NADPH的影响

目的: 研究褪黑素对肝内胆汁淤积大鼠的干预作用及氧化应激干扰的相关机制。方法:腹腔注射α-萘异硫氰酸酯建立肝内胆汁淤积大鼠模型，给予褪黑素灌胃进行干预治疗。检测肝功能生化指标，进行肝组织病理学观察以评价模型及药效；TBA法、微板法检测肝组织匀浆中一氧化氮（NO）、丙二醛（MDA）、谷胱甘肽（GSH）的含量，全自动定量易Western检测（WES）、逆转录-聚合酶链反应（RT-PCR）检测还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）蛋白/核酸在肝组织中的表达水平。 结果:血清肝功能生化检测结果及肝脏病理组织切片观察结果显示α-萘异硫氰酸酯较为成功地复制了肝内胆汁淤积大鼠模型，褪黑素对该模型有较好的干预作用。与正常组相比，模型组大鼠肝组织GSH显著降低（P<0.05），NO、MDA显著升高（P<0.01）；经褪黑素干预后，与模型组相比，褪黑素组大鼠肝组织GSH显著升高（P<0.05），NO、MDA显著降低（P<0.01）；各组大鼠肝组织NADPH蛋白/核酸表达无统计学差异（P>0.05）。结论:褪黑素对肝内胆汁淤积大鼠干预作用显著，可有效改善肝内胆汁淤积大鼠的氧化应激状态；抗氧化应激机制为褪黑素干预肝内胆汁淤积大鼠模型的重要机制之一，具有进一步深入研究的意义。

Effects of melatonin on NO,MDA, GSH and NADPH expression in liver tissues of intrahepatic cholestasis rats

AIM:To investigate the intervention of melatonin on rat intrahepatic cholestasis model and related mechanisms of oxidative stress interference. METHODS: Intraperitoneal injection of alpha-naphthylisothiocyanate was used to establish rat model of intrahepatic cholestasis, and melatonin was administered intragastrically. The biochemical indexes of liver function were detected and liver histopathology were observed and evaluated, the content of NO, MDA, GSH in liver homogenate was detected by TBA and microplate test, the expression of NADPH protein and mRNA in liver homogenate was detected by WES and RT-PCR.RESULTS:The results of biochemical detection of liver function in rat serum and observation of liver pathological sections showed that alpha-naphthylisothiocyanate could successfully reproduce the intraahepatic cholestasis rat model, and melatonin had an ideal intervention effect on the model. Compared with the normal group, the GSH of liver tissue in the model group was significantly lower than that in the normal group (P<0.05), the expression levels of NO and MDA increased significantly (P<0.01). After treatment with melatonin, GSH expression was significantly elevated in the liver tissue of the melatonin group as compared with the model group (P<0.05) while NO, MDA expression levels are significantly reduced (P<0.01). There was no significant difference in NADPH protein/mRNA expression levels in liver tissue of each group of rats (P>0.05).CONCLUSION:The effect of melatonin on intrahepatic cholestasis in rats is significant, which can effectively improve the oxidative stress status of rats with intrahepatic cholestasis. The anti-oxidative stress mechanism may be considered as one of the important mechanisms of melatonin intervention in the Intrahepatic cholestasis rat model, which has important significance for further research.