# Natural Protective Mechanisms of Cucumis callosus Leaves in Escherichia Species-Induced Urinary Tract Infection: An Integrated In Silico and In Vivo Study

## TL;DR

Cucumis callosus leaves show antibacterial and protective effects against urinary tract infections caused by Escherichia species, both in computer models and animal studies.

## Contribution

This is the first study to investigate Cucumis callosus leaf fractions for treating Escherichia-induced UTIs using an integrated in silico, in vitro, and in vivo approach.

## Key findings

- Chloroform fraction F1 of Cucumis callosus leaves showed potent antibacterial activity against uropathogenic Escherichia species.
- Molecular docking confirmed the binding of F1 compounds to bacterial resistance enzymes like AmpC β-lactamase and carbapenemases.
- In vivo studies showed F1 reduced bacterial load and inflammation in infected rats, with histopathological evidence of tissue protection.

## Abstract

Leaves of Cucumis callosus, traditionally employed in Ayurvedic medicine for the treatment of urinary disorders, were investigated in depth for their therapeutic potential against bacterially induced urinary tract infection (UTI) for the first time. The present work is the first to explore the antibacterial activity of C. callosus leaf fractions with an integrative in silico, in vitro, and in vivo approach. Through bioassay-guided fractionation, the chloroform fraction (F1) was identified as the most active, exhibiting potent activity against Uropathogenic Escherichia spp. species. Liquid chromatography–mass spectrometry (LC-MS) analysis of F1 revealed the presence of bioactive compounds, including stigmasterol, 1,2,3,4-tetrahydroisoquinoline, lactose, hydroxy(mesityl)acetic acid, and 2,4-di-tert-butylphenol. Molecular docking studies validated the strong binding affinities of these compounds for bacterial resistance enzymes, including AmpC β-lactamase and carbapenemases, thereby providing plausible mechanisms of antimicrobial action. In vivo studies carried out on female rats infected with Escherichia spp. species revealed a dose-dependent reduction in bacterial load, with a significant decrease in urinary tract inflammation upon F1 administration. Histopathological evaluation confirmed the protective effect, with reduced epithelial damage and inflammation in bladder tissues. These findings indicate significant antibacterial and tissue-protective effects of the C. callosus leaf fraction F1, supporting its ethnomedicinal use and establishing it as a promising phytotherapeutic agent for the treatment of urinary tract infections.