Sodium-Glucose Cotransporter-2 Inhibitor Suppresses Endoplasmic Reticulum Stress and Oxidative Stress in Diabetic Nephropathy Through Nrf2 Signaling: A Clinical and Experimental Study.

Diabetic nephropathy (DN), a severe complication of type 2 diabetes mellitus (T2DM), is marked by heightened endoplasmic reticulum stress (ERS) and oxidative stress (OS) due to protein misfolding and free radical generation. We investigated the sodium-glucose co-transporter-2 inhibitor (SGLT2i), canagliflozin (Cana), in alleviating ERS and OS in DN patients and THP-1 cells under hyperglycemic condition. A total of 120 subjects were divided into four groups, with 30 subjects in each group: healthy controls, T2DM individuals, DN patients receiving standard treatment, and those treated with Cana. The control group had no history of diabetes, cardiovascular or renal diseases, or other comorbidities. Cana was administered at doses of either 100 or 300 mg per day based on the estimated glomerular filtration rate (eGFR) value of DN individuals, with a mean follow-up of 6 months. Additionally, THP-1 monocytes were exposed to HGM (33.3 mM glucose with a cytokine cocktail of TNF-α and IFN-γ at 50 ng/mL each) to evaluate the relative levels of ERS, OS markers, and nuclear factor erythroid 2-related factor 2 (Nrf2), the transcription factor regulating cellular redox, which is downregulated in diabetes. Our results revealed that ERS markers GRP78 and PERK, as well as OS markers TXNIP and p22phox, were elevated in both DN patients and HGM-treated THP-1 monocytes and were reduced by Cana intervention. Furthermore, Cana regulated the phosphorylation of Nrf2, Akt, and EIF2α in HGM-treated monocytes. In conclusion, our findings highlight the role of Cana in activating Nrf2, thereby attenuating ERS and OS to mitigate DN progression.

Pterostilbene accelerates wound healing response in diabetic mice through Nrf2 regulation.

Pterostilbene (PTS), known for its diverse beneficial effects via Nuclear factor erythroid-2 related factor (Nrf2) activation, holds potential for Diabetic Foot Ulcer (DFU) treatment. However, PTS-mediated Nrf2 regulation in diabetic wounds has yet to be elucidated. We used IC21 macrophage-conditioned media to simulate complex events that can influence the fibroblast phenotype using L929 cells during the wound healing process under a hyperglycemic microenvironment. We found that PTS attenuated fibroblast migration and alpha-smooth muscle actin (α-SMA) levels and hypoxia-inducible factor- 1 alpha (HIF1α). Furthermore, we demonstrated that wounds in diabetic mice characterized by impaired wound closure in a heightened inflammatory milieu, such as the NOD-like receptor P3 (NLRP3) and intercellular adhesion molecule 1 (ICAM1), and deficient Nrf2 response accompanying lowered Akt signaling and heme oxygenase1 (HO1) expression along with the impaired macrophage M2 marker CD206 expression, was rescued by administration of PTS. Such an elicited response was also compared favorably with the standard treatment using Regranex, a commercially available topical formulation for treating DFUs. Our findings suggest that PTS regulates Nrf2 in diabetic wounds, triggering a pro-wound healing response mediated by macrophages. This insight holds the potential for developing targeted therapies to heal chronic wounds, including DFUs.

Histone deacetylase inhibitors as antidiabetic agents: Advances and opportunities.

The loss of function or dysfunction of ß-cells in the pancreas, attributed to the development of diabetes, involve alterations in genetic and epigenetic signatures. Recent evidences highlight the pathophysiological role of histone deacetylases (HDACs) in type 1 and type 2 diabetes. Indeed, most HDAC members have been linked to critical pathogenic events in diabetes, including redox imbalance, endoplasmic reticulum (ER) homeostasis perturbation, onset of oxidative stress and inflammation, which ultimately deteriorate ß-cell function. Accumulating evidence highlights the inhibition of HDACs as a prospective therapeutic strategy. Several chemically synthesized small molecules have been investigated for their specific ability to inhibit HDACs (reffered as HDAC inibitors) in various experimental studies. This review provides insights into the critical pathways involved in regulating different classes of HDACs. Further, the intricate signaling networks between HDACs and the stress mediators in diabetes are also explored. We exhaustively sum up the inferences from various investigations on the efficiency of HDAC inhibitors in managing diabetes and its associated complications.

Metabolic reprogramming and immune regulation in viral diseases.

The recent outbreak and transmission of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide and the ensuing coronavirus disease 2019 (COVID-19) pandemic has left us scrambling for ways to contain the disease and develop vaccines that are safe and effective. Equally important, understanding the impact of the virus on the host system in convalescent patients, healthy otherwise or with co-morbidities, is expected to aid in developing effective strategies in the management of patients afflicted with the disease. Viruses possess the uncanny ability to redirect host metabolism to serve their needs and also limit host immune response to ensure their survival. An ever-increasingly powerful approach uses metabolomics to uncover diverse molecular signatures that influence a wide array of host signalling networks in different viral infections. This would also help integrate experimental findings from individual studies to yield robust evidence. In addition, unravelling the molecular mechanisms harnessed by both viruses and tumours in their host metabolism will help broaden the repertoire of therapeutic tools available to combat viral disease.

Dysregulation of Nrf2 redox pathway in macrophages under diabetic microenvironment.

In the present study, we characterized the aberration in Nrf2 signaling in macrophages under a hyperglycemic microenvironment that reflects diabetic wounds in vitro and studied the effect of an Nrf2 activator pterostilbene (PTS) in these experimental conditions. Macrophages were exposed to pro-inflammatory cytokines TNFα and IFNγ with (HG+) or without high glucose (NG+) followed by the treatment with or without PTS. Western blotting was undertaken to assess the Nrf2 translocation from cytosol to nucleus followed by its downstream and upstream mediators, heme oxygenase-1 and Akt, respectively, the latter via phosphorylation. Quantitative PCR was also carried out to check the expression of macrophage mannose receptor CD206. We found a 2-fold reduction in the activation of Nrf2 in the HG+ group at 24 h compared to NG+, which was significantly improved by the treatment with PTS. Reduction in the levels of heme oxygenase-1 and phosphorylation of Akt in the HG+ group was also ameliorated by PTS. Furthermore, the gene expression of CD206 that was significantly reduced in the HG+ group was also restored by PTS treatment. The disruption of Nrf2 signaling in macrophages in a hyperglycemic microenvironment in vitro may indeed reflect diabetic wounds, as opposed to other non-diabetic wounds.

Crosstalk between endoplasmic reticulum stress and oxidative stress in the progression of diabetic nephropathy.

Increasing evidence in substantiating the roles of endoplasmic reticulum stress, oxidative stress, and inflammatory responses and their interplay is evident in various diseases. However, an in-depth mechanistic understanding of the crosstalk between the intracellular stress signaling pathways and inflammatory responses and their participation in disease progression has not yet been explored. Progress has been made in our understanding of the cross talk and integrated stress signaling network between endoplasmic reticulum stress and oxidative stress towards the pathogenesis of diabetic nephropathy. In this present study, we studied the crosstalk between the endoplasmic reticulum stress and oxidative stress by understanding the role of protein disulfide isomerase and endoplasmic reticulum oxidase 1α, a key player in redox protein folding in the endoplasmic reticulum. We had recruited a total of 90 subjects and divided into three groups (control (n = 30), type 2 diabetes mellitus (n = 30), and diabetic nephropathy (n = 30)). We found that endoplasmic reticulum stress markers, activating transcription factor 6, inositol-requiring enzyme 1α, protein kinase RNA-like endoplasmic reticulum kinase, C/EBP homologous protein, and glucose-regulated protein-78; oxidative stress markers, thioredoxin-interacting protein and cytochrome b-245 light chain; and the crosstalk markers, protein disulfide isomerase and endoplasmic reticulum oxidase-1α, were progressively elevated in type 2 diabetes mellitus and diabetic nephropathy subjects. The association between the crosstalk markers showed a positive correlation with endoplasmic reticulum stress and oxidative stress markers. Further, the interplay between endoplasmic reticulum stress and oxidative stress was investigated in vitro using a human leukemic monocytic cell line under a hyperglycemic environment and examined the expression of protein disulfide isomerase and endoplasmic reticulum oxidase-1α. DCFH-DA assay and flow cytometry were performed to detect the production of free radicals. Further, phosphorylation of eIF2α in high glucose-exposed cells was studied using western blot. In conclusion, our results shed light on the crosstalk between endoplasmic reticulum stress and oxidative stress and significantly contribute to the onset and progression of diabetic nephropathy and therefore represent the major therapeutic targets for alleviating micro- and macrovascular complications associated with this metabolic disturbance. Graphical abstract.

Macrophage mediation in normal and diabetic wound healing responses.

PURPOSE: The failure in timely healing of wounds is a central feature in chronic wounds that leads to physiological, psychological and economic burdens. Macrophages have been demonstrated to have various functions in wounds including host defense, the promotion and resolution of inflammation, the removal of apoptotic cells and tissue restoration following injury. Accumulated evidence suggests that macrophage dysfunction is a component of the pathogenesis of non-healing wounds. While the overall signaling cascades have been well understood, their complex interplay and a detailed characterization of events that are disrupted in chronic wounds have still not emerged satisfactorily. METHODS: The existing literature was reviewed to summarize the regulation of macrophage polarization in wound closure and dysregulation in non-healing wounds. Further, the review also underscored the role of Nrf2 in promoting macrophage-mediated regulation in wound responses and in particular, macrophage involvement in iron homeostasis that is impaired in chronic wounds such as in diabetes. RESULTS: The mechanisms involved in the reprogramming of macrophage subtypes in chronic wounds are still emerging. Furthermore, treating non-healing wounds has increasingly been shifting focus from generic treatments to the development of targeted therapies. Increasing evidence suggests the need for modeling wound tissue in vitro which may very well serve a critical aspect to characterize the relevant factors that sustain chronic wounds in vivo such as the constant iron overload at the wound site from recurrent infection and bleeding. CONCLUSION: The development of targeted therapies and also developing a reliable means to monitor assisted healing of chronic wounds are two major goals to be pursued. In addition, identifying molecular targets that can regulate macrophages to aid tissue restoration in chronic wounds would serve the crucial step in realizing both aforementioned goals.