Dihydroquercetin alleviates mitochondrial dysfunction and inhibits NLRP3-mediated pyroptosis in primary hepatocytes from chronic liver failure patients.

Objective

To investigate the protective effects of dihydroquercetin (DHQ) on mitochondrial function in primary hepatocytes from patients with chronic liver failure (CLF) and discuss the underlying molecular mechanism.

Methods

Blood samples from 15 CLF patients and 15 healthy subjects were retrospectively analyzed. Primary hepatocytes were isolated using a modified two-step collagenase perfusion method. The optimal concentration of DHQ (20 μmol/L) was determined using the Cell Counting Kit-8 (CCK-8) assay. Cells were divided into a control group, a plasma group, and a DHQ intervention group. The assessed indicators included cell viability (CCK-8), apoptosis rate (flow cytometry), inflammatory factors (IL-1β, IL-6, TNF-α; ELISA), oxidative stress markers (MDA, SOD, GSH-Px), mitochondrial function (JC-1 staining, mtDNA/nDNA ratio) and related protein expression (Western blot).

Results

DHQ adjunctive therapy reduced serum circulating mitochondrial DNA levels in patients with CLF, increased serum SOD2 and Humanin expression, inhibited activation of the NLRP3-mediated pyroptotic pathway, improved remission rates of decompensated complications, and showed favorable safety and tolerability in patients with end-stage liver diseases. In vitro, hepatocyte viability was markedly decreased in the CLF group, with increased apoptosis, elevated IL-1β, IL-6, TNF-α, and MDA levels, as well as declined SOD and GSH-Px activities. DHQ reduced apoptosis, up-regulated mitochondrial functional markers (SOD2, Humanin), restored JC-1 red/green fluorescence ratios, inhibited NLR family pyrin domain-containing 3 (NLRP3) inflammasome activation, and suppressed the caspase-1/IL-1β axis.

Conclusion

DHQ exerts multidimensional protective effects on hepatocytes, providing a novel strategy and clinical evidence for mitochondria-targeted CLF therapy.