Harnessing the Power of Bifidobacterium Isolates: Mitigating Aflatoxin B1 Toxicity in HT-29 Cell Lines and BALB/c Mice for Therapeutic Applications

Atieh Darbandi ¹ Naser Kakavandi ² Soheila Shahroodian ² Malihe Talebi ^2*

• ¹ Shahed University, Tehran, Tehran, Iran
• ² Iran University of Medical Sciences, Tehran, Iran

This study assessed the toxicity, mutagenicity, and genotoxicity of five probiotic candidate Bifidobacterium strains isolated from healthy individuals to introduce them as pharmacological supplements that inhibit aflatoxin-induced cancer cells to treat liver cancer.Genotoxicity evaluations of Bifidobacterium strains were conducted using three key methodologies: the Ames test with Salmonella typhimurium for bacterial assays, an in vitro mammalian chromosome aberration test for examining chromosomal anomalies in cell cultures, and an in vivo mouse micronucleus assay to assess genotoxic potential in the peripheral blood of mice. This study also examined strains' protection against aflatoxin B1 (AFB1) toxicity in human mucus-secreting cells.In the chromosomal aberration experiment, the strains were non-toxic. Moreover, single gavage of Bifidobacterium isolates at 2000, 1000, and 500 mg/kg b.w./day to BALB/c mice caused no mortality, clinical complaints, body weight alterations, or morphological abnormalities and had low acute toxicity. Probiotics lowered anticancer efficacy in HT-29 cells treated with Bifidobacterium isolates and exposed to AFB1 (P<0.05). They also lowered Bax and caspase-3 and boosted Bcl-2 and tight junction protein (Occludin and Zonula occludens-1) expression in AFB1-treated HT-29 cells. Inflammatory marker Interleukin-6 and nutrition transport genes decreased in this group. In animal studies, Bifidobacterium treatment reduced AFB1-induced liver damage and boosted the Nuclear factor erythroid Factor 2 and Glutathione S-transferase A3 production. These Bifidobacterium isolates also decreased cytochrome P450 (1A2 /3A4) expression.At the quantities studied, Bifidobacterium isolates did not indicate mutagenic potential in vitro by bacterial reverse mutation, clastogenicity, or in vivo. Probiotics may prevent mycotoxin cytotoxicity by protecting gut cell architecture and function and animal models.