Azilsartan Reduced TNF-α and IL-1β Levels, Increased IL-10 Levels and Upregulated VEGF, FGF, KGF, and TGF-α in an Oral Mucositis Model

Abstract

Oral mucositis (OM) is a common complication of treatments for head and neck cancer, particularly radiotherapy with or without chemotherapy. OM is characterised by oral erythema, ulceration, and pain. The aim of this study was to evaluate the effect of azilsartan (AZT), an angiotensin II receptor antagonist, on 5-fluorouracil (5-FU)-induced oral mucositis (OM) in Syrian hamsters. OM was induced by the intraperitoneal administration of 5-FU on experimental days 1 (60mg/Kg) and 2 (40mg/Kg). Animals were pretreated with oral AZT (1, 5, or 10 mg/kg) or vehicle 30 min before 5-FU injection and daily until day 10. Experimental treatment protocols were approved by the Animal Ethics Committee Use/CEUA (Number 28/2012) of the UFRN. Macroscopic analysis and cheek pouch samples were removed for histopathologic analysis. Myeloperoxidase (MPO), Malonyldialdehyde (MDA), interleukin-1 beta (IL-1β), interleukin-10 (IL-10), and tumour necrosis factor-alpha (TNF-α) were analysed by Enzyme Linked Immuno Sorbent Assay (ELISA). Vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), keratinocyte growth factor (KGF), and transforming growth factor (TGF)-α were measured by immunohistochemistry. Analysis of variance followed by Bonferroni’s test was used to calculate the means of intergroup differences (p ≤ 0.05). Treatment with 1 mg/kg AZT reduced levels MPO (p<0.01), MDA (p<0.5) and histological inflammatory cell infiltration, and increased the presence of granulation tissue. AZT treatment at 1 mg/kg reduced the TNF-α (p<0.05) and IL-1β (p<0.05) levels, increased the cheek pouch levels of IL-10 (p<0.01), and upregulated VEGF, FGF, KGF, and TGF-α. Administration of AZT at higher doses (5 and 10 mg/kg) did not significantly reverse the OM. AZT at a dose of 1 mg/kg prevented the mucosal damage and inflammation associated with 5-FU-induced OM, increasing granulation and tissue repair.

Mechanisms of Normal Tissue Injury From Irradiation

“…Deficiency of TNF-α in a lung injury model was sufficient to prevent symptoms of radiation pneumonitis 70 . TNF-α has also been implicated in radiation mucositis, enteritis, and dermatitis 71–75 . The clinical availability of agents targeting TNF-α to treat rheumatologic disorders provides a unique opportunity for clinical translation, and will likely enhance investigation into this molecule and its pathway.…”

Role of the route of leukotrienes in an experimental model of oral mucositis induced by 5-fluorouracil

“…It was found that the IL-10 levels in hamsters with 5-FU-induced OM were higher than those in healthy animals 27 . The use of COX-2 inhibitors and cys-LTs inhibitors (at doses of 10 and 20 mg/ kg/d for 10 d) also did not significantly reduce the expression of this cytokine.…”

“…The OM model presents increased expression of TNF-α and IL1-β at its inflammatory peak (day 10) 13,[24][25][26][27][28] . The present study corroborates such results, demonstrating a significant increase in TNF-α and IL1-β expression in the positive (5-FU) control group.…”

Effect of Gold Nanoparticle on 5-Fluorouracil-Induced Experimental Oral Mucositis in Hamsters

“…The tissue blocks were cut with 5-µm microtome, fixed, and stained on histological slides with dewaxing in xylol, hydration in alcohol baths (100% to 70%), stained with hematoxylin and eosin, dehydrated in alcohol baths (70% to 100%), clarified with xylol, and slides were assembled for analysis by light microscopy (40× Nikon E200 LED, São Paulo, Brazil). Oral mucosa were classified into score: Score 1: Normal epithelium and connective tissue without vasodilation, absent or discrete cellular infiltration, or absence of hemorrhagic areas, ulcerations, or abscesses; score 2: Discrete areas of vasodilation or re-epithelialization, slight inflammatory infiltration with mononuclear prevalence, and absence of hemorrhagic areas, edema, ulcerations, or abscesses; score 3: Moderate vasodilation, areas of epithelial degeneration, inflammatory infiltration with neutrophil prevalence, presence of hemorrhagic areas, edema, and eventual ulceration, and absence of abscesses; and score 4: Severe vasodilation and inflammatory infiltrate with neutrophils [26].…”

“…The plates were read at 490 nm by an ELISA plate reader (Polaris, Celer, Belo Horizonte, Brazil). To obtain the results in pg/mL, the standard curve was obtained according to the manufacturer's instructions for each antibody [24,26].…”