The Combination of Orlistat and Spirulina Attenuates Angiogenesis, Proliferation, Invasion, and Oxidative Stress in a C57bl/6 Mice Model of Hepatocellular Carcinoma Publisher



Shiri H ; Meshkani R ; Taheri M A ; Noorbakhsh F ; Allameh A ; Panahi G
Source: Algal Research Published:2026

Abstract

Hepatocellular carcinoma (HCC) is the leading cause of cancer-related mortality worldwide. Drug resistance, systemic toxicity, and poor outcomes limit current first-line sorafenib therapy, which calls for novel therapeutic strategies. Orlistat and Spirulina have each demonstrated independent anticancer effects. HCC was induced in male C57BL/6 J mice using a DEN/CCl₄ protocol. Animals were randomly allocated to six groups for 4 weeks of treatment: control, HCC model, Spirulina (500 mg/kg/day), orlistat (200 mg/kg/day), sorafenib (30 mg/kg/day), and the Spirulina-orlistat combination. Glypican-3 as a specific marker of HCC evaluated via immunohistochemistry. Angiogenesis was evaluated by assessing HIF-1α protein levels, Vegf, and Vegf receptor gene expression. Proliferation was assessed using Ki-67 immunofluorescence. Zymography was performed to evaluate invasiveness by measuring MMP-2/MMP-9 activity and Timp1 gene expression. Oxidative stress was assessed by measuring SOD/CAT activity, TAC, MDA, and Nrf2 gene expression. The combination of orlistat and Spirulina significantly reduced tumorigenic parameters, including glypican-3 expression, angiogenesis (decreased HIF-1α, Vegf, and Vegf receptor), proliferation (reduced Ki-67), invasion (decreased MMP-2/9 activity and increased Timp1 expression), and improved oxidative stress (elevated CAT/SOD activity, upregulation of Nrf2 gene expression, and reduced MDA). These effects were comparable to those of sorafenib across all assessed parameters. Moreover, the combination reduced orlistat's pro-oxidative liability while restoring sorafenib's effect on hepatic antioxidant balance. This combination exerts anti-tumor effects by targeting angiogenesis, proliferation, invasion, and oxidative stress, with efficacy comparable to sorafenib while uniquely surpassing it in restoring hepatic redox homeostasis. This combination strategy may serve as an alternative or adjunctive therapy for HCC; however, further investigation is needed to establish its synergistic effects via single-agent and combination studies with sorafenib, to explore additional HCC-related pathways, and through in vitro experiments. © 2026 Elsevier B.V.