3-methyladenine prevents sleep disturbance-induced liver injury by suppressing autophagy and endoplasmic reticulum stress

Abstract:

Objective To explore the protective effect and corresponding mechanisms of 3-methyladenine（3-MA） on liver injury induced by sleep disturbance.Methods Thirty SD mice were randomly divided into three groups，control group， modeling group，and 3-MA group， with 10 mice in each group. After the end of exposure， the weight changes in mice were measured and compared among the groups before and after the experiments. After the mice were sacrificed， the following experiments were made. ①The level changes of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were detected by ELISA method. ②HE staining was conducted to observe the histomorphological changes of the liver. ③Autophagosomes were analyzed using transmission electron microscope. ④TUNEL method was used to evaluate the apoptosis level of hepatocytes in each group and calculate the apoptosis index. ⑤RT-PCR and Western blot methods were used to detect the mRNA and expression levels of autophagy-related proteins，such as microtubule-associated protein light-chain-3 （LC3-Ⅱ），Beclin1，glucose-regulated protein 78（GRP78），and Caspase-3.Results The animal experiments showed that sleep disturbance could induce the liver damage，and which was accompanied with the increase of autophagy and endoplasmic reticulum stress（ERS）. Compared with the control group，the body weight increment of the model group decreased significantly，while the morphological structure of hepatocytes remarkably changed. Meantime，the levels of ALT and AST，apoptosis index，autophagosomes and the expression of related marker proteins increased respectively（P<0.05）. In contrast with the model group，the morphological structure，serum ALT and AST，apoptosis index，autophagosomes and the expression of related marker proteins were improved（P<0.05）.Conclusion Sleep disturbance exposure can induce liver damage in SD rats，and then further activate autophagy and ERS signaling pathways by up-regulating the protein expression and mRNA level of LC3-Ⅱ，Beclin1 and GRP78. 3-MA can antagonize autophagy and ERS signaling pathways to protect hepatocytes and resist apoptosis.