Regulatory effect of PINK1/Parkin axis on mitophagy in isoniazide-induced hepatocellular injury.

Abstract

Patients with tuberculosis, who are treated with long-term high-dose combined use of anti-tuberculosis drugs, can cause many adverse reactions such as liver damage, but the mechanism is still unclear. Although phosphatase and tensin homolog induced kinase 1 (PINK1)/Parkin axis might participate in the process of liver damage through regulating mitochondrial autophagy and oxidative stress in liver cells. However, the association between the mitochondrial autophagy regulated by the PINK1/Parkin axis and the liver injury induced by anti-tuberculosis drugs is unknown. This study aims to explore the mechanism of PINK1/Parkin signal axis in regulating hepatocellular injury induced by anti-tuberculosis drugs through mitochondrial autophagy, and to provide new therapeutic targets for the patients with liver damage caused by anti-tuberculosis drugs. Mouse hepatocytes AML-12 were treated with isoniazide (INH) to induce liver injury. The mRNA and protein levels of PINK1, Parkin and autophagy associated factors were detected by real-time PCR and Western blotting in the normal AML-12 cells (control group), the AML-12 cells treated by INH (model group) and the AML-12 cells treated with INH and salidroside together (intervention group). Meantime, ELISA kit was used to detect the level of reactive oxygen species (ROS) in AML-12 cells, and the cell morphology and injury was observed by HE staining and transmission electron microscope (TEM). Compared with the control group, the mRNA (all P <0.01) and protein levels (all P <0.05) of PINK1, Parkin and microtubule associated protein 1 light chain 3 (LC3) were significantly decreased, the ubiquitination level was significantly decreased ( P <0.05), and the ROS level was increased ( P <0.05), and the hepatocellular cell showed obvious damage in the model group. Compared with the model group, the expression of PINK1, Parkin and LC3 were significantly increased in the intervention group (all P <0.05), as well as the ubiquitination level was also increased ( P <0.05). The ROS level mediated by mitochondria was decreased ( P <0.05). The results of HE staining and TEM showed that the cell status was improved and the damage of hepatocytes was significantly attenuated. In this study, the mechanism of mitochondrial autophagy mediated by PINK1/Parkin signal axis may be related to INH-induced injury in liver cells and it can regulate the damage state in vitro, indicating that Parkin is a potential molecular target for the prediction, diagnosis and treatment for liver injury induced by anti-tuberculosis drugs.