IN SILICO AND IN VITRO EVALUATION OF ENROFLOXACIN ON AFLATOXIN B1-INDUCED CYTOTOXICITY

Abstract

Additional therapeutics are required to minimize the toxicity of aflatoxin B1 (AFB1) in animals due to the drawbacks of mycotoxin binders. The cytochrome P450 enzyme system is necessary for the metabolic activation of AFB1 before it produces the cytotoxic AFB1-exo-8, 9-epoxide (AFBO). It is already proven that enrofloxacin (ENR) has been shown to substantially decrease the activity of the cytochrome P450 enzymes. To understand how ENR affects AFB1-induced cytotoxicity, it is important to highlight this context. In this present study, molecular docking was performed between ENR with CYP3A4 protein, apoptotic proteins (Bax, caspase 3, caspase 8, fas L, MAPK1), and catalase. To assess the effect of ENR on AFB1-induced cytotoxicity by MTT assay and trypan blue dye exclusion techniques, the mammalian simulative Vero cell lines were used in different treatment groups as AFB1 alone, ENR alone, AFB1+ENR, and AFB1+silymarin (known cell protective agent). This was done to further evaluate the in silico analysis. ENR interacted more positively with all of the proteins (CYP3A4, Bax, catalase, caspase 3, caspase 8, MAPK1, and fas L), according to the findings of molecular docking studies. On the other hand, cytotoxicity caused by AFB1 is successfully prevented by ENR at a dose of 25 µg/mL. The results of the present study suggest that ENR primarily protects against cytotoxicity induced by AFB1 in Vero cells as the evidence of in silico studies revealed that ENR may protect AFB1-induced cytotoxicity by interacting with the CYP3A4 enzyme which is primarily required for activation AFB1.

Keywords

• aflatoxin b1
• enrofloxacin
• molecular docking
• apoptotic proteins
• in vitro study