Objective: To investigate the effect of
berberine on programmed
necrosis of hepatocytes induced by metabolic-associated
fatty liver disease (MAFLD) in mice and its related molecular mechanism. Methods: Twenty male C57BL/6N mice were randomly divided into four groups (n=5 in each group): control group (S),
fatty liver group (H),
berberine group(B), nuclear factor erythroid 2-related factor 2 inhibitor group (Nrf2), and
all-trans-retinoic acid (ATRA) group (A). Serum
alanine aminotransferase (ALT),
aspartate aminotransferase (AST),
lactate dehydrogenase (LDH),
triglycerides (TG), total
cholesterol (TC),
tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) concentrations were detected at the end of week 12 to calculate
fatty liver index (liver mass/body mass ratio). Liver tissue was stained with HE, Masson and
Oil Red O, and
SAF score was used to evaluate the degree of liver injury. The expression levels of hepatic programmed
necrosis-related
proteins, namely receptor-interacting
protein kinase 3 (RIPK3), phosphorylated mixed series
protease-like domain (p-MLKL) and Nrf2 were detected by Western blot method. One-way ANOVA was used for intragroup comparisons and
LSD-t tests were used for intergroup comparisons. Results: Compared with S group, H group serum ALT, AST, LDH, TG, TC, TNF-α, IL-1β levels and
fatty liver index were significantly increased. The liver tissue was filled with vacuolar-like changes and inflammatory cell infiltration. Numerous red lipid droplets were observed with
oil red O staining.
Collagen fiber
hyperplasia was evident with Masson staining.
SAF scores (6.60 ± 0.55 and 0.80 ± 0.45) were significantly increased. The expressions of RIPK3 and p-MLKL were up-regulated. Nrf2 level was relatively increased, and the differences were statistically significant (P < 0.05). Compared with H group,
berberine intervention group liver biochemical indexes,
lipid levels, pro-inflammatory mediator expression,
fatty liver index, and
SAF score were significantly reduced, and the expression of RIPK3 and p-MLKL were down-regulated, while Nrf2 levels were further increased, and the differences were statistically significant (P<0.05). Compared with B group, treatment with Nrf2 inhibitor had antagonized the protective effect of
berberine on
fatty liver. Serum ALT, AST, LDH, TG, TC and TNF-α, IL-1β levels,
fatty liver index, and
SAF scores were significantly increased and the expressions of RIPK3 and p-MLKL were relatively increased, and the differences were statistically significant (P < 0.05). Conclusion:
Berberine can significantly improve the metabolic-associated
fatty liver disease injury in mice, and its mechanism is related to activation of Nrf2 and inhibition of programmed
necrosis of hepatocytes.