In silico molecular docking and in vivo evidence of whey protein concentrate-mediated protection against thioacetamide-induced intestinal toxicity in male albino rats via modulation of oxidative stress, inflammation, apoptosis, and fibrosis

Abstract

Oxidative stress is a foremost cause in the etiology and progression of numerous illnesses. Thioacetamide (TA) has been demonstrated to promote oxidative stress and has been shown to be harmful in a variety of organs. The antioxidant activity of whey protein concentrate (WPC) has generated a lot of interest since it can help with the nutritional treatment of chronic disorders. The present study examined WPC’s antioxidant qualities, evaluating any potential defenses against TA-induced intestinal damage in rats. Forty rats were equally divided into four groups and treated for 5 days per week over 3 weeks: untreated control; TA (thioacetamide 100 mg/kg/day i.p.); WPC (whey protein 300 mg/kg/day oral); and WPC + TA (WPC orally followed 2 h later by TA i.p.). The results showed that TA treatment dramatically increased levels of nitric oxide and malondialdehyde while significantly decreasing levels of glutathione and activity of antioxidant enzymes (superoxide dismutase plus catalase) in intestinal tissue. Furthermore, TA injection showed higher values in apoptotic markers (Bcl-2 and Bax) and inflammatory indicators (IL-1 β and TNF- α ), and reduced expression of genes such as ZO-1 and HO-1 in intestinal tissue. Using a molecular docking study, the potential binding mechanisms of the antioxidant peptide with TGF- β and Keap1were examined. Additionally, there were notable immunopositive reactions for NF-kB and α . SMA, as well as significant histological changes, increased collagen fiber deposition, and duodenal goblet cell hyperplasia were observed in the TA group. However, WPC pretreatment significantly decreased intestinal tissue’s oxidative stress, pro-inflammatory, apoptotic, and fibrotic indicators, hence reducing TA-induced intestinal damage, suggesting WPC can be a useful and economical feed option that improves intestinal health.