Interventional Effects and Potential Mechanisms of Tibetan Medicine Shiwei Dida Capsules on Metabolic Dysfunction-Associated Steatohepatitis Based on Zebrafish Model

OBJECTIVE To evaluate the intervention effects of Tibetan medicine Shiwei Dida capsules on egg yolk powder-induced metabolic dysfunction-associated steatohepatitis(MASH) based on zebrafish model, and to preliminarily explore its possible mechanism.

METHODS Wild-type AB strain zebrafish larvae at 5 days post-fertilization(5 dpf) were used for the determination of maximum tolerated concentration. Zebrafish were randomly divided into blank group, model group, positive control group, and low-, medium-, and high-dose Shiwei Dida capsule groups(37.5, 75, 150 μg·mL⁻¹). Drug intervention was carried out simultaneously with egg yolk powder induction. After treatment, whole-mount oil red O staining was used to evaluate lipid deposition in the liver region, and HE staining was applied to observe histopathological changes of liver tissue. The levels of ALT, AST, TC, TG, HDL-C, and LDL-C were measured. The mRNA expression levels of PPARG, Fasn, LPL, TNF-α, and IL-1β were determined by qRT-PCR.

RESULTS Shiwei Dida capsules showed no apparent toxicity to zebrafish larvae at concentrations of 150 μg·mL⁻¹ and below. Compared with the blank group, the integrated optical density(IOD) values of oil red O staining in the liver region, the levels of ALT, AST, TC, TG, LDL-C, as well as the relative mRNA expression levels of PPARG, Fasn, TNF-α, and IL-1β were significantly increased in the model group, while the HDL-C level and relative LPL mRNA expression level were significantly decreased(P<0.05 or P<0.001). Hepatocellular swelling, tissue structural disorder and lipid droplet-like vacuolation could be observed pathologically. Compared with the model group, the IOD values of oil red O staining, levels of ALT, AST, TC and LDL-C were decreased and HDL-C level was increased in each dose group of Shiwei Dida capsules. The TG levels were reduced in medium- and high-dose groups of Shiwei Dida capsules(P<0.05 or P<0.01), while there was no statistically significant difference in TG level in the low-dose group. The expression of IL-1β mRNA was downregulated(P<0.001) and the expression of LPL mRNA was upregulated(P<0.05 or P<0.001) in all dose groups of Shiwei Dida capsules. The medium- and high-dose groups of Shiwei Dida capsules could downregulate the mRNA expression of PPARG and TNF-α(P<0.05, P<0.01 or P<0.001), and only the high-dose group could downregulate Fasn mRNA expression(P<0.05).

CONCLUSION Shiwei Dida capsules exert an explicit protective effect against egg yolk powder-induced MASH in zebrafish, and its effect may be related to improving lipid metabolic disorder and inhibiting inflammatory response.