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Published on: July 10, 2019
IQ-RKT Formulation mitigates cardiomyocytes injury by targeting AGEs-RAGE-ROS-dependent TRAF3IP2/JNK apoptotic nexus
Humera Jahan1, Urooba Fatima2, Sana Asad2
1Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan; Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore 54000, Pakistan.
Abstract:
Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality in diabetic populations. Elevated advanced glycation end products (AGEs) in diabetes foster the on-set, and progression of CVDs. However, the underlying AGEs-induced signaling nexus, involved in cardiomyocytes apoptosis, remain unexplored. Currently no anti-AGEs drug available to address CVDs in diabetes. Indeed, natural products remained a major sources of new medicine, and currently being focused in the drug development. The objective of this study was to explore the mechanism of AGEs-associated apoptotic pathway in cardiomyocytes. Additionally, to harness the medicinal properties of natural products, a newly developed formulation, comprised of rutin, kaempferol, and thymoquinone, named IQ-RKT, was characterized for its anti-apoptotic potential against AGEs-induced cardiotoxicity. We studied the role of MGO-, and glucose-AGEs in cardiomyocytes apoptosis under diabetic environment in H9c2 cells in vitro, as well as in SD diabetic rats in vivo. The inhibition of AGEs-induced apoptosis of cardiomyocytes was investigated by a treatment with IQ-RKT. Using H9c2 cells, as well as SD diabetic rats models in vivo, we found that AGEs-induced elevated levels of RAGE was reduced by a treatment with IQ-RKT. AGEs stimulate intracellular ROS generation, TRAF3 interacting protein 2 (TRAF3IP2) expression, and TRAF3IP2-dependent-JNK activation. TRAF3IP2/JNK causes transactivation of AP-1/NF-κB transcription factors. AGEs increase Bax, cytochrome c, and activate caspase-3, and suppress anti-apoptotic Bcl-2. IQ-RKT significantly inhibited this apoptotic pathway, and tilted the balance towards anti-apoptosis. Moreover, IQ-RKT decreased lipid peroxidation, cardiac injury, and glycooxidative biomarkers in the plasma of diabetic rats. Interestingly, this effect of IQ-RKT was independent of hyperglycemic environment in diabetic rats. The IQ-RKT also appeared as a stable formulation at different pH, and temperature ranges. The study provides experimental evidences that AGE-RAGE axis is likely associated with cardiomyocytes death via ROS-dependent TRAF3IP2/JNK pathway. Therefore, targeting this pathway appeared to have therapeutic potential. The newly developed formulation IQ-RKT was identified as an effective anti-apoptotic cardioprotective agent to be further investigated through pre-clinical, and clinical studies.
Insights
Cardiovascular diseases in diabetes are linked to advanced glycation end products (AGEs). A natural product formulation, IQ-RKT, effectively reduced AGE-induced cardiomyocyte apoptosis via a novel signaling pathway.
Area of Science:
- Cardiovascular research
- Diabetology
- Natural product chemistry
Background:
- Cardiovascular diseases (CVDs) are a major complication in diabetic populations, significantly driven by elevated advanced glycation end products (AGEs).
- The specific signaling pathways through which AGEs induce cardiomyocyte apoptosis remain incompletely understood, and effective anti-AGE therapies for diabetic CVDs are lacking.
- Natural products offer a promising avenue for developing novel therapeutic agents.
Purpose of the Study:
- To elucidate the mechanism of AGE-induced apoptosis in cardiomyocytes.
- To evaluate the cardioprotective potential of a novel natural product formulation, IQ-RKT (comprising rutin, kaempferol, and thymoquinone), against AGEs in a diabetic context.
- To investigate the role of the AGE-RAGE axis and downstream signaling in AGE-mediated cardiomyocyte death.
Main Methods:
- In vitro studies using H9c2 cells and in vivo studies using Sprague-Dawley diabetic rats were employed to model AGE-induced cardiovascular damage.
- The effects of IQ-RKT on AGE-induced apoptosis, receptor for AGEs (RAGE) levels, reactive oxygen species (ROS) generation, TRAF3IP2/JNK pathway activation, and apoptotic markers (Bax, Bcl-2, caspase-3) were assessed.
- Biomarkers of lipid peroxidation, cardiac injury, and glycooxidative stress were measured in plasma from diabetic rats.
Main Results:
- IQ-RKT treatment significantly reduced AGE-induced RAGE elevation, ROS generation, and TRAF3IP2-dependent JNK activation in cardiomyocytes.
- The formulation inhibited the AGEs-triggered activation of transcription factors AP-1 and NF-κB, leading to increased Bax, cytochrome c, and caspase-3, and decreased Bcl-2.
- In vivo, IQ-RKT decreased lipid peroxidation, cardiac injury, and glycooxidative biomarkers in diabetic rats, independent of hyperglycemia, and demonstrated formulation stability.
Conclusions:
- The AGE-RAGE axis promotes cardiomyocyte death through a ROS-dependent TRAF3IP2/JNK signaling pathway.
- The natural product formulation IQ-RKT exhibits significant anti-apoptotic and cardioprotective effects by targeting this pathway.
- IQ-RKT represents a promising therapeutic candidate for managing diabetic cardiovascular complications, warranting further clinical investigation.
