Withaferin-A in Ameliorating the Effects of High Glucose on Inflammatory and Phagocytic Response of Mouse Macrophages.

Whyte, Trae; Sinha, Satyesh K.

Rapidly increasing rates of diabetes mellitus (DM) throughout the world represent an emerging epidemic with profound consequences including diabetic nephropathy (DN). Studies indicate that mφ-mediated inflammation correlates with the development of DN. Macrophages exhibit pro- (M1) and anti-inflammatory (M2) phenotypes. Therefore, in the present study, we tested our hypothesis that high glucose suppresses the M2 phenotype and phagocytosis, leading to aberrant cytokine release, and that withaferin-A (an anti-inflammatory molecule) will reduce the pro-inflammatory response of macrophages. We cultured J-774A.1 macrophage-like cells (ATCC) in RPMI 1640. After reaching 70- 80% confluence, the cells were serum starved for 18 hours. Cells were then treated with Dglucose (5, and 25mM) for 24h. We extracted the total protein from the cells to measure the expression level of arginase-1, TGF- beta, LC3 and GPNMB by way of the western blot process. In a second set of experiments, we treated the cells with withaferin-A. In these cells, we will measure the level of secreted pro-inflammatory cytokines in culture media using ELISA. Preliminary data under our ongoing experiments indicate that high glucose treated macrophages exhibited reduced levels of markers of M2 phenotype (Arginase-1, and TGF- beta) and phagocytosis (GPNMB, and LC3). We are planning to conduct the experiments related to the excretion of pro-inflammatory cytokines in high glucose with and without withaferin-A. Our results indicate that high glucose modulates the expression of markers of M2 phenotype and phagocytosis in macrophages. We are continuing our experiments to confirm the effects of withaferin-A on the secretion of pro-inflammatory cytokines in high glucose.

LACTONES; MACROPHAGES; INFLAMMATION; HYPERGLYCEMIA; DIABETIC nephropathies; LABORATORY mice

Journal of Health Disparities Research & Practice, 2016, Vol 9, p38

2166-5222

Academic Journal