Resveratrol Upregulates Antioxidant Factors Expression and Downmodulates Interferon-Inducible Antiviral Factors in Aging

Abstract

Immunosenescence, a process with a dysfunctional immune response that may favor infection is associated with an increase in inflammatory responses mediated by proinflammatory cytokines, characteristic of inflammaging. Aging and immunosenescence have a relationship relating to oxidative stress and inflammaging. Therefore, natural antioxidant compounds could be candidates for the control of the oxidative process. Our purpose was to evaluate the effect of resveratrol (Resv) on the antioxidant, antiviral, and anti-inflammatory responses induced by toll-like receptors (TLRs) 3, 4, and 7/8 agonists stimulation on peripheral blood mononuclear cells (PBMCs) of elderly and healthy female individuals (63-82 years old) and young and healthy female individuals (21-31 years old). Our data show that Resv may upregulate antioxidant factor expression, such as catalase (CAT) and SIRT1, in response to TLR4 and TLR7/8 agonists, similarly in both young and aged groups. Moreover, the Resv anti-inflammatory effect was detected by inhibiting IL-1β, TNF-α, and IL-10 secretion levels, as well as by the chemokines CCL2 and CCL5, induced by TLR4 and TLR7/8 stimulation. Curiously, Resv decreased antiviral genes, such as MxA, STING, and IRF7 expression, possibly by reducing the inflammatory effects of interferon-induced genes. Taken together, our results demonstrate the ability of Resv to stimulate antioxidant factors, leading to a downmodulation of the inflammatory response induced by innate immune stimulation. These findings point out Resv as a strategy to control the upregulation of inflammatory response, even in elderly individuals.

Keywords: aging; antioxidant; antiviral; inflammaging; innate immunity; resveratrol; toll-like receptors.

Figure 1

Resv downmodulates antiviral factors upon TLR3 activation. (A) Heatmap of antioxidant and antiviral factors expressed by PBMC stimulated with TLR agonist 3 (POLY(I:C)) and addition of Resv. The young group is colored in green, and the elderly group is colored in yellow. Red shade gene expression means above the row average, and blue shade expression means below the average. Z-score represents subtract mean, divided by standard deviation. (B) Comparison of the constitutive gene expression of young healthy volunteers and elderly healthy volunteers by qPCR. The relative expression of the targets was calculated in comparison to the amplification of the constitutive gene, GAPDH, and in comparison, to the non-stimulated situation. N = 9–10 individuals per group. Data are expressed as median and interquartile range. Paired Wilcoxon test: * p < 0.05, ** p < 0.001 (stimulated vs. unstimulated).

Figure 2

Resv upregulates CAT/SIRT1 expressions while decreasing antiviral factors upon TLR4 activation. (A) Heatmap of antioxidant and antiviral factors expressed by PBMCs stimulated with TLR agonist 4 (LPS) and addition of Resv. The young group is colored in green, and the elderly group is colored in yellow. Red shade gene expression means above the row average, and blue shade expression means below the average. Z-score represents subtract mean, divided by standard deviation. (B) Comparison of the constitutive gene expression of young healthy volunteers and elderly healthy volunteers by qPCR. The relative expression of the targets was calculated in comparison to the amplification of the constitutive gene, GAPDH, and in comparison to the non-stimulated situation. N = 9–10 individuals per group. Data are expressed as median and interquartile range. Paired Wilcoxon test: * p < 0.05, ** p < 0.001 (stimulated vs. unstimulated).

Figure 3

Resv upregulates CAT expressions while decreasing antiviral factors upon TLR7/8 activation. (A) Heatmap of antioxidant and antiviral factors expressed by PBMC stimulated with TLR agonists 7 and 8 (CL097) and addition of Resv (N = 20). The young group is colored in green, and the elderly group is colored in yellow. Red shade gene expression means above the row average, and blue shade expression means below the average. Z-score represents subtract mean, divided by standard deviation. (B) Comparison of the constitutive gene expression of young healthy volunteers and elderly healthy volunteers by qPCR. The relative expression of the targets was calculated in comparison to the amplification of the constitutive gene, GAPDH, and in comparison to the non-stimulated situation. N = 9–10 individuals per group. Data are expressed as median and interquartile range. Paired Wilcoxon test: * p < 0.05, ** p < 0.001 (stimulated vs. unstimulated).

Figure 4

Proinflammatory cytokines induced by TLR4 and TLR7/8 stimulation were inhibited by Resv. PBMCs were incubated for 24 h with CL097 (2.5 µg/mL), LPS (1 µg/mL), POLY(I:C) (10 µg/mL), and Resv (100 µM). The production of cytokines IL-1β, TNF-α, IFN-γ, and IL-10 was assessed by flow cytometry. N = 9–10 individuals per group. Data expressed in the median and interquartile range. Unpaired Mann–Whitney test: # p < 0.05 (young vs. elderly). Wilcoxon paired test: * p < 0.05; ** p < 0.01 (stimulated vs. unstimulated).

Figure 5

CCL2 and CCL5 induced by TLR4 and TLR3 stimulation were inhibited by Resv. PBMCs were incubated for 24 h with CL097 (2.5 µg/mL), LPS (1 µg/mL), and POLY(I:C) (10 µg/mL) in the presence of Resv (100 µM). CCL2 and CCL5 chemokines were assessed by flow cytometry. N = 9–10 individuals per group. Data expressed as median and interquartile range. One-way ANOVA test: * p < 0.05; ** p < 0.01. Unpaired Mann Whitney test: # p < 0.05 (stimulated vs. unstimulated).