Interference With the AMPKα/mTOR/NLRP3 Signaling and the IL-23/IL-17 Axis Effectively Protects Against the Dextran Sulfate Sodium Intoxication in Rats: A New Paradigm in Empagliflozin and Metformin Reprofiling for the Management of Ulcerative Colitis

Mahmoud E Youssef; Eslam E Abd El-Fattah; Amir M Abdelhamid; Hanan Eissa; Eman El-Ahwany; Noha A Amin; Helal F Hetta; Mohamed H Mahmoud; Gaber El-Saber Batiha; Naglaa Gobba; Ahmed Gaafar Ahmed Gaafar; Sameh Saber

doi:10.3389/fphar.2021.719984

Interference With the AMPKα/mTOR/NLRP3 Signaling and the IL-23/IL-17 Axis Effectively Protects Against the Dextran Sulfate Sodium Intoxication in Rats: A New Paradigm in Empagliflozin and Metformin Reprofiling for the Management of Ulcerative Colitis

Front Pharmacol. 2021 Aug 16:12:719984. doi: 10.3389/fphar.2021.719984. eCollection 2021.

Authors

Affiliations

¹ Department of Pharmacology, Faculty of Pharmacy, Delta University for Science and Technology, Gamasa, Egypt.
² Department of Biochemistry, Faculty of Pharmacy, Delta University for Science and Technology, Gamasa, Egypt.
³ Department of Clinical Pharmacology, Faculty of Medicine, Mansoura University, Mansoura, Egypt.
⁴ Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt.
⁵ Department of Hematology, Theodor Bilharz Research Institute, Giza, Egypt.
⁶ Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut, Egypt.
⁷ Department of Internal Medicine, College of Medicine, University of Cincinnati, Cincinnati, OH, United States.
⁸ Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia.
⁹ Department of Pharmacology and Therapeutics, Faculty of Veterinary Medicine, Damanhour University, Damanhour, Egypt.
¹⁰ Department of Pharmacology and Toxicology, College of Pharmacy, Misr University for Science and Technology, 6th of October City, Egypt.
¹¹ Department of Pharmacology and Toxicology, Faculty of Pharmacy, Port-Said University, Port-Said, Egypt.

Abstract

Empagliflozin and metformin are widely used for the treatment of type 2 diabetes. These drugs showed marked anti-inflammatory effects in different animal models via enhancing AMPK activity. Yet, the protective anti-inflammatory effects of their combination against ulcerative colitis have not been previously investigated. The current study aimed to explore the potential of empagliflozin/metformin combination to mitigate the DSS-induced rat colitis model. The modulating effects of empagliflozin and metformin on the AMPK/mTOR/NLRP3 axis and T cell polarization were delineated. In this study, distal colons were examined for macroscopic and microscopic pathological alterations. ELISA, qRT-PCR, and immunohistochemistry techniques were applied to detect proteins and cytokines involved in AMPK/mTOR/NLRP3 axis and T Cell polarization. Oral administration of empagliflozin (10 mg/kg/day) and metformin (200 mg/kg/day) combination alleviated colitis as revealed by the reduced disease activity index, macroscopic damage index, colon weight/length ratio, and histopathologic scoring values. Interestingly, empagliflozin/metformin combination significantly enhanced AMPK phosphorylation and depressed mTOR and NLRP3 expression leading to a subsequent reduction in caspase-1 cleavage and inhibition of several inflammatory cytokines, including IL-1β, and IL-18. Reduced mTOR expression and reduced IL-6 levels led to a reduction in Th17 cell polarization and maintenance. Together, the current study reveals that the protective effects of empagliflozin and metformin against DSS-induced colitis are fundamentally mediated via enhancing AMPK phosphorylation. Since adult humans with diabetes mellitus are at greater risk for developing inflammatory bowel diseases, clinical application of empagliflozin/metformin combination represents a novel therapeutic approach for treating diabetic patients with ulcerative colitis.

Keywords: AMPK; IL-23/1L-17 axis 4; NLRP3; dextran sulfate sodium; empagliflozin; mTOR; metformin; ulcerative colitis.