Myricitrin pretreatment ameliorates mouse liver ischemia reperfusion injury.

BACKGROUND

Myricitrin has been reported to exert protective effects on liver diseases, but the protective effects of myricitrin against liver ischemia reperfusion (I/R) injury and the underlying mechanisms remain unexplored. This study aimed to investigate the effects of myricitrin on liver I/R injury and elucidate the underlying mechanisms.

METHODS

Mice were pretreated with myricitrin before liver I/R injury modeling. The mice were pretreated with either myricitrin or vehicle prior to liver ischemia. Some mice were further pretreated with the PI3K inhibitor LY294002. Liver tissues and blood samples were collected after 6 h of reperfusion. The degree of liver damage was determined by the serum levels of alanine aminotransferase (ALT), aspartate transaminase (AST), and lactic dehydrogenase (LDH) and histological examinations. The tumour necrosis factor-α (TNF-α), interleukin--1β (IL-1β), IL-4 and IL-10 expression levels were assessed by qRT-PCR and enzyme-linked immunosorbent assays (ELISAs). Serum superoxide dismutase (SOD) activity, catalase (CAT) activity, and contents of malondialdehyde (MDA), glutathione (GSH) and nitric oxide (NO) contents were measured. Western blotting and caspase-3 activity were conducted to determine the effect of myricitrin on apoptosis. The expression levels of proliferation related genes (Cyclin D1 and Cyclin E1) were determined by qRT-PCR and western-blotting. The expression of p-Akt, p-mTOR and p-eNOS in liver tissue were investigated by western-blotting.

RESULTS

Myricitrin not only significantly decreased the ALT, AST and LDH levels but also reduced the necrotic areas in the liver tissue compared with liver I/R injury group. In addition, myricitrin pretreatment alleviated liver injury by inhibiting the inflammatory response and suppressing oxidative stress. Western blotting and caspase-3 activity revealed that myricitrin inhibited liver I/R induced-apoptosis. Myricitrin promoted hepatocyte proliferation following liver I/R injury by upregulating the expression levels of Cyclin D1 and Cyclin E1. Further experiments indicated that the myricitrin pretreatment increased nitric oxide (NO) production by activating the PI3K/Akt signaling pathway. However, myricitrin triggered the hepatocyte proliferation and NO synthase activation was blocked by LY294002.

CONCLUSION

These results demonstrate that myricitrin alleviates liver I/R injury by suppressing oxidative stress, the inflammatory response, and apoptosis, improving liver proliferation and upregulating p-eNOS expression.