Oxyhydrogen nanobubbles modulate β-catenin and CD8⁺ in a rat model of hepatocarcinoma

Abstract

Hepatocellular carcinoma (HCC) continues to be a significant contributor to cancer-associated fatalities, highlighting the worldwide necessity for efficacious treatment approaches. This research assessed the impact of oxyhydrogen nanobubbles (HHOnbs) on β-catenin expression and CD8⁺ T-cell infiltration in a rat model of HCC caused by diethylnitrosamine (DEN) and carbon tetrachloride (CCl₄), while also contrasting their dose-dependent effects with Lenvatinib. Male Wistar rats were divided into six categories: negative control, positive control, HHOnbs given 12 or 24 times, and Lenvatinib provided 3 or 5 times. We evaluated β-catenin expression and the infiltration of CD8⁺ T-cells by immunohistochemical methods. HCC induction elevated β-catenin expression and augmented CD8⁺ T-cell infiltration. HHOnb treatment dose-dependently reduced β-catenin expression by 59.98% and 89.87% and increased CD8⁺ T-cell infiltration by 49.23% and 129.23% following 12 and 24 administrations, respectively. HHOnbs showed stronger β-catenin suppression, whereas Lenvatinib elicited greater CD8⁺ T-cell infiltration. These findings indicate that HHOnbs can suppress oncogenic signaling and enhance antitumor immune responses in experimental HCC. Future studies should assess their long-term efficacy and safety profiles and investigate combination therapies, as the present study did not evaluate these parameters.