Abstract 214: Role Of Sirtuin 6 In Macrophage Polarization In Diabetes

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
G V Velmurugan ◽  
Suresh K Verma ◽  
Alexander R Mackie ◽  
Erin E Vaughan ◽  
Raj kishore ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Glucose ◽  
Macrophage Polarization ◽  
Experimental Studies ◽  
Raw 264.7 Cells ◽  
Lv Function ◽  
Mouse Bone Marrow ◽  
Intramyocardial Injection ◽  
Disease States ◽  
High Glucose Condition

Clinical and experimental studies provide evidence that metabolic and inflammatory pathways are functionally interconnected to cardiovascular diseases. Dynamic changes in macrophage activation [classical M1 activation (promote inflammation) or alternative M2 activation (promote wound healing)], in response to various signals, modulate tissue pathology in diabetes. Sirtuin 6 (SIRT6), a NAD-dependent nuclear deacetylase plays an important role in genomic stability, cellular metabolism, stress response and aging. However, the mechanism by which SIRT6 activity affects macrophage function in diabetes is still unclear. Mouse bone marrow-derived macrophages (BMM) exposed to high glucose (HG, 25mM D-glucose) showed reduced expression of SIRT6 as compared to low glucose (LG, 5mM D-glucose)- and osmotic control (OC, 5mM D-glucose+20mM D-mannitol)-treated cells. SIRT6 knockdown in RAW 264.7 cells exaggerated inflammatory response in macrophages exposed to high glucose (HG) and in contrast, IL4-induced alternative macrophage (M2) phenotype was defective in SIRT6 deficient BM-macrophages under high glucose condition. SIRT6 protein expression was low in failing (MI-induced) and diabetes-affected hearts. Interestingly, mice receiving intramyocardial injection of SIRT6-deficient macrophages showed further deterioration in LV function, post-MI. Taken together, these data highlighting a role for SIRT6 in regulating both M1 and M2 polarization might have broad implications for numerous inflammatory disease states including insulin resistance and wound healing.

Abstract 11059: Role of Sirtuin 6 in Macrophage Polarization and Cardiac Repair in Diabetes

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Rajarajan A Thandavarayan ◽  
Darukeshwara Joladarashi ◽  
Sahana S Babu ◽  
Garikipati V Srikanth ◽  
Alexander R Mackie ◽  
...  
Keyword(s):  
Macrophage Polarization ◽  
Experimental Studies ◽  
Peritoneal Macrophages ◽  
Left Ventricular ◽  
Cardiac Repair ◽  
Raw 264.7 Cells ◽  
Mouse Bone Marrow ◽  
Macrophage Phenotype ◽  
Intramyocardial Injection ◽  
M2 Phenotype

Clinical and experimental studies provide evidence that metabolic and inflammatory pathways are functionally interconnected to cardiovascular diseases. Dynamic changes in macrophage activation [classical M1 activation (promote inflammation) or alternative M2 activation (promote wound healing)], in response to various stress signals, modulate cardiac physiopathology in diabetes. Sirtuin 6 (SIRT6), a NAD-dependent nuclear deacetylase plays an important role in genomic stability, cellular metabolism, stress response and aging. However, the mechanism by which SIRT6 activity affects macrophage phenotype and cardiac function in diabetes is still unexplored. Mouse bone marrow-derived macrophages (BMM) exposed to high glucose (HG, 25mM D-glucose) showed reduced expression of SIRT6 as compared to low glucose (LG, 5mM D-glucose)- and osmotic control (OC, 5mM D-glucose+20mM D-mannitol)-treated cells, associated with increased expression of proinflammatory cytokine and transcription factors (NFkb, c-JUN, FOXO, SP1 and STAT1). In addition, SIRT6 level was reduced in peritoneal macrophages of both diabetic models (streptozotocin-induced and db/db mice) as compared to non-diabetic mice. SIRT6 knockdown in RAW 264.7 cells exaggerated inflammatory response when exposed to HG. In contrast, IL-4-induced increase in mRNA expression of macrophage M2 phenotype markers like Arg1, Chi4l4, Retnla and IRS-2, but not IRS-1 expression was repressed suggesting that alternative macrophage (M2) phenotype was defective in SIRT6 deficient BM-macrophages under HG condition. SIRT6 protein expression was low in myocardial infarction-induced (MI) and diabetes-affected hearts. Interestingly, mice receiving intramyocardial injection of SIRT6-deficient macrophages showed further deterioration in left ventricular function, post-MI. Taken together, these data highlight a role for SIRT6 in regulating the balance of M1/M2 polarization, therefore, modulate macrophage mediated cardiac repair and regeneration in numerous inflammatory disease states including diabetes

scholarly journals Protective Effects of Thalidomide on High-Glucose-Induced Podocyte Injury through In Vitro Modulation of Macrophage M1/M2 Differentiation

2020 ◽  
Vol 2020 ◽  
pp. 1-14
Author(s):  
Hui Liao ◽  
Yuanping Li ◽  
Xilan Zhang ◽  
Xiaoyun Zhao ◽  
Dan Zheng ◽  
...  
Keyword(s):  
High Glucose ◽  
Macrophage Polarization ◽  
Mannose Receptor ◽  
Raw 264.7 Cells ◽  
Protective Effects ◽  
Activated Macrophages ◽  
Podocyte Injury ◽  
Protein Expressions ◽  
Activated Macrophage

Objective. It has been shown that podocyte injury represents an important pathological basis that contributes to proteinuria and eventually leads to kidney failure. High glucose (HG) activates macrophage polarization, further exacerbating HG-induced podocyte injury. Our previous study on diabetic nephropathy rats indicated that thalidomide (Tha) has renoprotective properties. The present study explored the effects of Tha on mRNA and protein expressions of inducible nitric oxide synthase (iNOS), tumor necrosis factor- (TNF-) α, mannose receptor (CD206), and arginase- (Arg-) 1 in HG-activated macrophages. iNOS and TNF-α are established as markers of classically activated macrophage (M1). CD206 and Arg-1 are regarded as markers of alternatively activated macrophages (M2). During the experiment, the supernatants of (HG)-treated and (Tha)-treated macrophages, designated as (HG) MS and (Tha) MS, were simultaneously collected and processed. TNF-α and interleukin- (IL-) 1β levels as well as protein expressions of nephrin and podocin in HG, (HG) MS, and (Tha) MS-cultured podocytes were evaluated. The results showed that compared to the 11.1 mM normal glucose (NG), the 33.3 mM HG-cultured RAW 264.7 cells exhibited upregulated iNOS and TNF-α mRNAs and protein expressions, and downregulated CD206 and Arg-1 expressions significantly (p<0.05). Tha 200 μg/ml suppressed iNOS and TNF-α, and promoted CD206 and Arg-1 expressions significantly compared to the HG group (p<0.05). Furthermore, (HG) MS-treated podocytes showed an increase in TNF-α and IL-1β levels and a downregulation in nephrin and podocin expression significantly compared to NG-treated and HG-treated podocytes (p<0.05). The (Tha 200 μg/ml) MS group exhibited a decrease in TNF-α and IL-1β level, and an upregulation in nephrin and podocin expressions significantly compared to the (HG) MS group (p<0.05). Our research confirmed that HG-activated macrophage differentiation aggravates HG-induced podocyte injury in vitro and the protective effects of Tha might be related to its actions on TNF-α and IL-1β levels via its modulation on M1/M2 differentiation.

P6297Genetic mutation of galectin-3 altered the temporal evolution of macrophage polarization and healing affecting the post myocardial infarct remodeling in mice

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
G Gonzalez ◽  
P D Cassaglia ◽  
F Penas ◽  
N L Martinez ◽  
C Bettazza ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mrna Expression ◽  
Temporal Evolution ◽  
Macrophage Polarization ◽  
Weight Ratio ◽  
Lv Function ◽  
M2 Macrophage ◽  
Infarct Zone ◽  
Tnf Α ◽  
Galectin 3

Abstract Background Myocardial infarction (MI) is a dynamic process that leads to ventricular remodeling (VR) and largely to heart failure (HF). Previous studies established that Galectin-3 (Gal-3) is highly increased in the infarct zone from the beginning of MI and also that it is a prognostic marker of HF. Purpose We aimed to study the effects of genetic deletion of Gal-3 on macrophage (MΦ) infiltration, cytokines expression, fibrosis and MMP-2 activity as well as VR and function after MI in mice. Methods Male C57BL/6J and Gal-3 KO mice were subjected to permanent coronary ligature or sham. At 1 week post-MI LV function and VR were studied by echocardiography. We also studied in the infarct zone: 1) F4/80+ MΦ infiltration by flow cytometry; 2) M2 macrophage polarization by detection of mannose receptor (MR) and chitinase-3-like protein-3 (YM1) phenotype markers by rt-qPCR; 3) mRNA expression of TNF-α, IL-6, IL-10 and TGF-β; 4) MMP-2 activity by zymography and 5) fibrosis by histology. Results Results are expressed as X±SEM; *p<0.05 C57 MI vs Gal-3 KO MI. After 1 week post-MI, the pulmonary congestion assessed by the lung weight/body weight ratio (mg/g) was 9±0.4, 8±0.5 and 9±0.4 in C57 sham, Gal-3 KO sham and C57 MI, respectively, nevertheless it was severely increased to 15±1.2* in infarcted Gal-3 KO mice. MΦ infiltration, cytokine expression and MMP-2 activity in the infarct zone are shown in the table. Table 1 Groups F4/80+ MR YM1 TNF-α IL-6 IL-10 TGF-β MMP-2 MΦ (%) (A.U.) (A.U.) (A.U.) (A.U.) (A.U.) (A.U.) (A.U.) C57 MI 5.6±0.9 (8) 0.5±0.2 (3) 1.7±0.4 (3) 26±0.3 (3) 1.7±0.5 (3) 0.7±0.03 (3) 1.8±0.5 (3) 1±0.1 (5) Gal-3 KO MI 2.6±0.4* (8) 1.8±0.4*(8) 4.2±0.4* (8) 0.4±0.5* (8) 6.7±0.9 * (8) 2.8±0.3 * (8) 0.5±0.2 * (8) 2±0.4* (6) A.U.: Arbitrary units. In MR, YM1, TNF-α, IL-6, IL-10 and TGF-β represent mRNA expression. In MMP-2 represent gelatinolytic activity. Number of samples is shown between parentheses. After 1 week post-MI, LV end diastolic dimension was increased from 4.4±0.1 to 4.8±0.2* at the same time that ejection fraction (%) was significantly reduced from 47±2 to 38±3* in C57+MI (13) and Gal-3 KO+MI (16) respectively. Collagen concentration in the infarct zone was significantly reduced from 30±1.2% (6) to 17±0.5* % (8) in C57 and Gal-3 KO respectively. Conclusion(s) Gal-3 is an essential regulatory factor for the early wound healing since it regulates the dynamics of the reparative process through the phenotypic profile of MΦ, the pro- and anti-inflammatory cytokines expression and fibrosis along the temporal evolution of MI in mice. The deficit of Gal-3 diminished the infiltration of MΦ altering its phenotypic polarization and consequently, the dynamics of the wound healing as well as aggravating the functional and structural evolution of cardiac remodeling. Acknowledgement/Funding Argentine Agency for Promotion of Science and Technology (PICT 2014-2320), University of Buenos Aires (UBACyT 20020170100619BA)

Differential responses of bone to angiotensin II and angiotensin(1-7): beneficial effects of ANG(1-7) on bone with exposure to high glucose

2021 ◽  
Vol 320 (1) ◽  
pp. E55-E70
Author(s):  
Nan-Nan Sha ◽  
Jia-Li Zhang ◽  
Christina Chui-Wa Poon ◽  
Wen-Xiong Li ◽  
Yue Li ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Glucose Level ◽  
High Glucose ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Ang Ii ◽  
Mineral Density ◽  
High Glucose Condition ◽  
High Glucose Level ◽  
Glucose Condition

Osteoporosis, diabetes, and hypertension are common concurrent chronic disorders. This study aimed to explore the respective effects of angiotensin II (ANG II) and angiotensin(1-7) [ANG(1-7)], active peptides in the renin-angiotensin system, on osteoblasts and osteoclasts under high-glucose level, as well as to investigate the osteo-preservative effects of ANG II type 1 receptor (AT1R) blocker and ANG(1-7) in diabetic spontaneously hypertensive rats (SHR). ANG II and ANG(1-7), respectively, decreased and increased the formation of calcified nodules and alkaline phosphatase activity in MC3T3-E1 cells under high-glucose level, and respectively stimulated and inhibited the number of matured osteoclasts and pit resorptive area in RANKL-induced bone marrow macrophages. Olmesartan and Mas receptor antagonist A779 could abolish those effects. ANG II and ANG(1-7), respectively, downregulated and upregulated the expressions of osteogenesis factors in MC3T3-E1 cells. ANG II promoted the expressions of cathepsin K and MMP9 in RAW 264.7 cells, whereas ANG(1-7) repressed these osteoclastogenesis factors. ANG II rapidly increased the phosphorylation of Akt and p38 in RAW 264.7 cells, whereas ANG(1-7) markedly reduced the phosphorylation of p38 and ERK under high-glucose condition. After treatments of diabetic SHR with valsartan and ANG(1-7), a significant increase in trabecular bone area, bone mineral density, and mechanical strength was only found in the ANG(1-7)-treated group. Treatment with ANG(1-7) significantly suppressed the increase in renin expression and ANG II content in the bone of SHR. Taken together, ANG II/AT1R and ANG(1-7)/Mas distinctly regulated the differentiation and functions of osteoblasts and osteoclasts upon exposure to high-glucose condition. ANG(1-7) could protect SHR from diabetes-induced osteoporosis.

scholarly journals 4-hexylresorcinol-induced protein expression changes in human umbilical cord vein endothelial cells as determined by immunoprecipitation high-performance liquid chromatography

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243975
Author(s):  
Yeon Sook Kim ◽  
Dae Won Kim ◽  
Seong-Gon Kim ◽  
Suk Keun Lee
Keyword(s):  
Wound Healing ◽  
Endothelial Cells ◽  
Protein Expression ◽  
Macrophage Polarization ◽  
Raw 264.7 Cells ◽  
Human Umbilical Cord ◽  
Raw 264.7 ◽  
Related Proteins ◽  
Umbilical Cord Vein ◽  
Human Umbilical Cord Vein

4-Hexylresorcinol (4HR) is used as a food preservative and an ingredient of toothpaste and cosmetics. The present study was performed using 233 antisera to determine the changes in protein expression induced by 4HR in human umbilical cord vein endothelial cells (HUVECs), and evaluated the 4HR-induced effects in comparison with previous results (Kim et al., 2019). Similar to RAW 264.7 cells, 4HR-treated HUVECs showed decreases in the expression of the proliferation-related proteins, cMyc/MAX/MAD network proteins, p53/RB and Wnt/β-catenin signaling, and they showed inactivation of DNA transcription and protein translation compared to the untreated controls. 4HR upregulated growth factors (TGF-β1, β2, β3, SMAD2/3, SMAD4, HGF-α, Met, IGF-1) and RAS signaling proteins (RAF-B, p38, p-p38, p-ERK-1, and Rab-1), and induced stronger expression of the cellular protection-, survival-, and differentiation-related proteins in HUVECs than in RAW 264.7 cells. 4HR suppressed NFkB signaling in a manner that suggests potential anti-inflammatory and wound healing effects by reducing M1 macrophage polarization and increasing M2 macrophage polarization in both cells. 4HR-treated HUVECs tended to increase the ER stress mediators by upregulating eIF2AK3, ATF4, ATF6, lysozyme, and LC3 and downregulating eIF2α and GADD153 (CHOP), resulting in PARP-1/AIF-mediated apoptosis. These results indicate that 4HR has similar effects on the protein expression of HUVECs and RAW 264.7 cells, but their protein expression levels differ according to cell types. The 4HR-treated cells showed global protein expression characteristic of anticancer and wound healing effects, which could be alleviated simultaneously by other proteins exerting opposite functions. These results suggest that although 4HR has similar effects on the global protein expression of HUVECs and RAW 264.7 cells, the 4HR-induced molecular interferences in those cells are complex enough to produce variable protein expression, leading different cell functions. Moreover, HUVECs have stronger wound healing potential to overcome the impact induced by 4HR than RAW 264.7 cells.

scholarly journals Inflammatory M1-like macrophages polarized by NK-4 undergo enhanced phenotypic switching to an anti-inflammatory M2-like phenotype upon co-culture with apoptotic cells

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Keizo Kohno ◽  
Satomi Koya-Miyata ◽  
Akira Harashima ◽  
Takahiko Tsukuda ◽  
Masataka Katakami ◽  
...  
Keyword(s):  
Wound Healing ◽  
Chronic Wounds ◽  
Macrophage Polarization ◽  
Phenotypic Switching ◽  
Apoptotic Cells ◽  
Phenotypic Switch ◽  
Inflammatory Phase ◽  
M1 Macrophage ◽  
Tnf Α ◽  
Anti Inflammatory

Abstract Background NK-4 has been used to promote wound healing since the early-1950s; however, the mechanism of action of NK-4 is unknown. In this study, we examined whether NK-4 exerts a regulatory effect on macrophages, which play multiple roles during wound healing from the initial inflammatory phase until the tissue regeneration phase. Results NK-4 treatment of THP-1 macrophages induced morphological features characteristic of classically-activated M1 macrophages, an inflammatory cytokine profile, and increased expression of the M1 macrophage-associated molecules CD38 and CD86. Interestingly, NK-4 augmented TNF-α production by THP-1 macrophages in combination with LPS, Pam3CSK4, or poly(I:C). Furthermore, NK-4 treatment enhanced THP-1 macrophage phagocytosis of latex beads. These results indicate that NK-4 drives macrophage polarization toward an inflammatory M1-like phenotype with increased phagocytic activity. Efferocytosis is a crucial event for resolution of the inflammatory phase in wound healing. NK-4-treated THP-1 macrophages co-cultured with apoptotic Jurkat E6.1 (Apo-J) cells switched from an M1-like phenotype to an M2-like phenotype, as seen in the inverted ratio of TNF-α to IL-10 produced in response to LPS. We identified two separate mechanisms that are involved in this phenotypic switch. First, recognition of phosphatidylserine molecules on Apo-J cells by THP-1 macrophages downregulates TNF-α production. Second, phagocytosis of Apo-J cells by THP-1 macrophages and activation of PI3K/Akt signaling pathway upregulates IL-10 production. Conclusion It is postulated that the phenotypic switch from a proinflammatory M1-like phenotype to an anti-inflammatory M2-like phenotype is dysregulated due to impaired efferocytosis of apoptotic neutrophils at the wound site. Our results demonstrate that NK-4 improves phagocytosis of apoptotic cells, suggesting its potential as a therapeutic strategy to resolve sustained inflammation in chronic wounds.

scholarly journals High glucose levels inhibit focal adhesion kinase-mediated wound healing of rat peritoneal mesothelial cells

2003 ◽  
Vol 63 (2) ◽  
pp. 722-731 ◽  
Author(s):  
Masahito Tamura ◽  
Akihiko Osajima ◽  
Shingo Nakayamada ◽  
Hirofumi Anai ◽  
Narutoshi Kabashima ◽  
...  
Keyword(s):  
Wound Healing ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
High Glucose ◽  
Mesothelial Cells ◽  
Peritoneal Mesothelial Cells ◽  
Glucose Levels

scholarly journals Computational Investigation of Transmural Differences in Left Ventricular Contractility

2016 ◽  
Vol 138 (11) ◽  
Author(s):  
Hua Wang ◽  
Xiaoyan Zhang ◽  
Shauna M. Dorsey ◽  
Jeremy R. McGarvey ◽  
Kenneth S. Campbell ◽  
...  
Keyword(s):  
Myocardial Contractility ◽  
Ex Vivo ◽  
Experimental Studies ◽  
Contractile Force ◽  
Left Ventricular ◽  
Lv Function ◽  
Fe Model ◽  
Myocardial Layers ◽  
Best Fit

Myocardial contractility of the left ventricle (LV) plays an essential role in maintaining normal pump function. A recent ex vivo experimental study showed that cardiomyocyte force generation varies across the three myocardial layers of the LV wall. However, the in vivo distribution of myocardial contractile force is still unclear. The current study was designed to investigate the in vivo transmural distribution of myocardial contractility using a noninvasive computational approach. For this purpose, four cases with different transmural distributions of maximum isometric tension (Tmax) and/or reference sarcomere length (lR) were tested with animal-specific finite element (FE) models, in combination with magnetic resonance imaging (MRI), pressure catheterization, and numerical optimization. Results of the current study showed that the best fit with in vivo MRI-derived deformation was obtained when Tmax assumed different values in the subendocardium, midmyocardium, and subepicardium with transmurally varying lR. These results are consistent with recent ex vivo experimental studies, which showed that the midmyocardium produces more contractile force than the other transmural layers. The systolic strain calculated from the best-fit FE model was in good agreement with MRI data. Therefore, the proposed noninvasive approach has the capability to predict the transmural distribution of myocardial contractility. Moreover, FE models with a nonuniform distribution of myocardial contractility could provide a better representation of LV function and be used to investigate the effects of transmural changes due to heart disease.

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