Objective: The present study aimed to investigate the effect and molecular mechanism of the PKM2 small molecule agonist TEPP-46 on the development of methionine choline-deficient (MCD) diet-induced nonalcoholic steatohepatitis (NASH) in mice.
Materials and methods: In this study, C57BL/6 mice were fed an MCD diet for 15 days to establish a NASH model. The protein expression levels of pyruvate kinase M2 (PKM2), PKM1, hypoxia-inducible factor-1α (HIF-1α) and NLRP3 in liver Kupffer cells (KCs) were measured by Western blotting. Immunofluorescence analysis was used to analyze the nuclear translocation of PKM2 in KCs, and the levels of IL-1β and TNF-α in mouse serum and the cell polarization indexes were determined. The MCD diet-fed mice were injected with 30 mg/kg of TEPP-46 intraperitoneally every 5 days. After 15 days, the liver tissue and peripheral blood were collected for analysis.
Results: We found the NASH model was successfully established after the mice were fed an MCD diet for 15 days. MCD feeding promoted the expression of the PKM2 monomer/dimer and inhibited the expression of the PKM2 tetramer in KCs. Immunofluorescence analysis further confirmed that MCD feeding inhibited the nuclear translocation of PKM2. Besides, MCD feeding promoted the expression of HIF-1α and NLRP3 in KCs, promoted M1 KCs polarization and inhibited M2 KCs polarization. Intraperitoneal injection 30 mg/kg of TEPP-46 significantly inhibited the development of MCD diet-induced NASH, alleviated the pathological changes in the liver, improved liver function, promoted the expression of the PKM2 tetramer in KCs, and inhibited the expression of HIF-1α and NLRP3.
Conclusions: This study demonstrated that TEPP-46, a small molecule agonist of PKM2, may inhibit the nuclear translocation of PKM2 and the activation of KCs by promoting the expression of PKM2 tetramers in KCs, thus inhibiting the development of MCD diet-induced NASH in mice.