Limits...
Advanced Glycation End Products Enhance Macrophages Polarization into M1 Phenotype through Activating RAGE/NF-κB Pathway.

Jin X, Yao T, Zhou Z, Zhu J, Zhang S, Hu W, Shen C - Biomed Res Int (2015)

Bottom Line: The present study showed that AGEs significantly promoted macrophages to express IL-6 and TNF-α.AGEs significantly increased RAGE expression in macrophages and activated NF-κB pathway, and the aforementioned effects were partly abolished by administration of anti-RAGE antibody or NF-κB inhibitor PDTC.In conclusion, our results suggest that AGEs enhance macrophage differentiation into proinflammatory M1 phenotype at least partly via RAGE/NF-κB pathway activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, 1665 Kongjiang Road, Shanghai 200092, China ; Department of Cardiology, Central Hospital of Minhang District, 170 Xinsong Road, Shanghai 201199, China.

ABSTRACT
Atherosclerotic lesions are accelerated in patients with diabetes. M1 (classically activated in contrast to M2 alternatively activated) macrophages play key roles in the progression of atherosclerosis. Since advanced glycation end products (AGEs) are major pathogenic factors and active inflammation inducers in diabetes mellitus, this study assessed the effects of AGEs on macrophage polarization. The present study showed that AGEs significantly promoted macrophages to express IL-6 and TNF-α. M1 macrophage markers such as iNOS and surface markers including CD11c and CD86 were significantly upregulated while M2 macrophage markers such as Arg1 and CD206 remained unchanged after AGEs stimulation. AGEs significantly increased RAGE expression in macrophages and activated NF-κB pathway, and the aforementioned effects were partly abolished by administration of anti-RAGE antibody or NF-κB inhibitor PDTC. In conclusion, our results suggest that AGEs enhance macrophage differentiation into proinflammatory M1 phenotype at least partly via RAGE/NF-κB pathway activation.

No MeSH data available.


Related in: MedlinePlus

Role of AGEs on RAGE/NF-κB pathway. The cells were incubated with AGEs 500 μg/mL for different time. The p-p65 and total p65 protein were measured as MFI (mean fluorescence intensity) by FCM. The NF-κB pathway was activated by AGEs (a). After pretreatment with anti-RAGE antibody for 60 min, NF-κB was still activated by AGEs (b). NF-κB pathway activation was partly inhibited by anti-RAGE antibody (c). Each point represents the mean value ± SD of triplicate determinations, representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4465680&req=5

fig6: Role of AGEs on RAGE/NF-κB pathway. The cells were incubated with AGEs 500 μg/mL for different time. The p-p65 and total p65 protein were measured as MFI (mean fluorescence intensity) by FCM. The NF-κB pathway was activated by AGEs (a). After pretreatment with anti-RAGE antibody for 60 min, NF-κB was still activated by AGEs (b). NF-κB pathway activation was partly inhibited by anti-RAGE antibody (c). Each point represents the mean value ± SD of triplicate determinations, representative of three independent experiments.

Mentions: Next, we evaluated the effect of AGEs on activation of NF-κB pathway in macrophages. Because activation of NF-κB pathway is associated with phosphorylation of NF-κB, phospho-NF-κB-p65 (p-p65) and NF-κB-p65 (p65) were measured as mean fluorescence intensity (MFI) by FCM in different time points after AGEs stimulation. The p-p65 level increased within 15 min after treatment with AGEs. Moreover, the effect peaked at 60 min and decreased thereafter. In contrast, the total NF-κB-p65 (p65) level remained unchanged over the time after AGEs treatment. Furthermore, the p-p65/p65 ratio increased significantly shortly after AGEs stimulation, peaked at 60 min, and then decreased (Figure 6(a)). These data indicate that AGEs can activate the NF-κB pathway.


Advanced Glycation End Products Enhance Macrophages Polarization into M1 Phenotype through Activating RAGE/NF-κB Pathway.

Jin X, Yao T, Zhou Z, Zhu J, Zhang S, Hu W, Shen C - Biomed Res Int (2015)

Role of AGEs on RAGE/NF-κB pathway. The cells were incubated with AGEs 500 μg/mL for different time. The p-p65 and total p65 protein were measured as MFI (mean fluorescence intensity) by FCM. The NF-κB pathway was activated by AGEs (a). After pretreatment with anti-RAGE antibody for 60 min, NF-κB was still activated by AGEs (b). NF-κB pathway activation was partly inhibited by anti-RAGE antibody (c). Each point represents the mean value ± SD of triplicate determinations, representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4465680&req=5

fig6: Role of AGEs on RAGE/NF-κB pathway. The cells were incubated with AGEs 500 μg/mL for different time. The p-p65 and total p65 protein were measured as MFI (mean fluorescence intensity) by FCM. The NF-κB pathway was activated by AGEs (a). After pretreatment with anti-RAGE antibody for 60 min, NF-κB was still activated by AGEs (b). NF-κB pathway activation was partly inhibited by anti-RAGE antibody (c). Each point represents the mean value ± SD of triplicate determinations, representative of three independent experiments.
Mentions: Next, we evaluated the effect of AGEs on activation of NF-κB pathway in macrophages. Because activation of NF-κB pathway is associated with phosphorylation of NF-κB, phospho-NF-κB-p65 (p-p65) and NF-κB-p65 (p65) were measured as mean fluorescence intensity (MFI) by FCM in different time points after AGEs stimulation. The p-p65 level increased within 15 min after treatment with AGEs. Moreover, the effect peaked at 60 min and decreased thereafter. In contrast, the total NF-κB-p65 (p65) level remained unchanged over the time after AGEs treatment. Furthermore, the p-p65/p65 ratio increased significantly shortly after AGEs stimulation, peaked at 60 min, and then decreased (Figure 6(a)). These data indicate that AGEs can activate the NF-κB pathway.

Bottom Line: The present study showed that AGEs significantly promoted macrophages to express IL-6 and TNF-α.AGEs significantly increased RAGE expression in macrophages and activated NF-κB pathway, and the aforementioned effects were partly abolished by administration of anti-RAGE antibody or NF-κB inhibitor PDTC.In conclusion, our results suggest that AGEs enhance macrophage differentiation into proinflammatory M1 phenotype at least partly via RAGE/NF-κB pathway activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, 1665 Kongjiang Road, Shanghai 200092, China ; Department of Cardiology, Central Hospital of Minhang District, 170 Xinsong Road, Shanghai 201199, China.

ABSTRACT
Atherosclerotic lesions are accelerated in patients with diabetes. M1 (classically activated in contrast to M2 alternatively activated) macrophages play key roles in the progression of atherosclerosis. Since advanced glycation end products (AGEs) are major pathogenic factors and active inflammation inducers in diabetes mellitus, this study assessed the effects of AGEs on macrophage polarization. The present study showed that AGEs significantly promoted macrophages to express IL-6 and TNF-α. M1 macrophage markers such as iNOS and surface markers including CD11c and CD86 were significantly upregulated while M2 macrophage markers such as Arg1 and CD206 remained unchanged after AGEs stimulation. AGEs significantly increased RAGE expression in macrophages and activated NF-κB pathway, and the aforementioned effects were partly abolished by administration of anti-RAGE antibody or NF-κB inhibitor PDTC. In conclusion, our results suggest that AGEs enhance macrophage differentiation into proinflammatory M1 phenotype at least partly via RAGE/NF-κB pathway activation.

No MeSH data available.


Related in: MedlinePlus