Analysis of Plasma MicroRNAs as Predictors and Biomarkers of Aging and Frailty in Humans

Iryna Rusanova, María E Diaz-Casado, Marisol Fernández-Ortiz, Paula Aranda-Martínez, Ana Guerra-Librero, Francisco J García-García, Germaine Escames, Leocadio Mañas, Darío Acuña-Castroviejo, Iryna Rusanova, María E Diaz-Casado, Marisol Fernández-Ortiz, Paula Aranda-Martínez, Ana Guerra-Librero, Francisco J García-García, Germaine Escames, Leocadio Mañas, Darío Acuña-Castroviejo

Abstract

Although circulating microRNAs (miRNAs) can modulate gene expression and affect immune system response, little is known about their participation in age-associated frailty syndrome and sarcopenia. The aim of this study was to determine miRNAs as possible biomarkers of age and frailty and their correlation with oxidative and inflammatory state in human blood. Three inflammation-related miRNAs (miR-21, miR-146a, and miR-223) and one miRNA related with the control of melatonin synthesis (miR-483) were analyzed. Twenty-two healthy adults, 34 aged robust, and 40 aged fragile patients were selected for this study. The expression of plasma miRNAs was assessed by RT-qPCR; plasma cytokines (IL-6, IL-8, IL-10, and TNFα) were analyzed by commercial kits, and plasma advanced oxidation protein products (AOPP) and lipid oxidation (LPO) were spectrophotometrically measured. Fragile subjects had higher miR-21 levels than control subjects, whereas miR-223 and miR-483 levels increased at a similar extend in both aged groups. All cytokines measured increased in aged groups compared with controls, without differences between robust and fragile subjects. The fragile group had a TNFα/IL-10 ratio significantly higher than robust and control groups. Aged groups also had higher AOPP and LPO levels than controls. Women presented higher AOPP and LPO levels and increased expression of miR-483 compared with men. Positive correlations between miR-21 and AOPP and between miR-483 and IL-8 were detected. The expression of miR-21 and the TNFα/IL-10 ratio were correlated positively with the presence of frailty, which suggests that these markers can be considered as possible biomarkers for age-related frailty.

Figures

Figure 1
Figure 1
miRNA levels in control and aged groups (a)–(d). Relative expression of these miRNAs was calculated using the 2−ΔΔCt method. The expression levels were normalized against U6 snRNA and against the average of the control group (Ct, general normalization). Data are presented as means ± SEM. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001 versus the control group. Regression and correlation analysis with relative expression of miRNAs, calculated using analysis of the Spearman correlation coefficient. Only those miRNAs that had significant correlation are shown (e) and (f). C: control group; R: robust group; F: fragile group.
Figure 2
Figure 2
Plasma levels of IL-6, IL-8, TNFα, and IL-10 in control and aged groups of subjects ((a)–(d)). TNFα/IL-10 ratio in the three studied groups (e). Data are presented as means ± SEM. ∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001 versus the control group. #P < 0.05 versus the robust group.
Figure 3
Figure 3
Lipid peroxidation (LPO) and advanced oxidation protein product (AOPP) levels in plasma of subjects of three studied groups ((a) and (b)). LPO and AOPP levels in plasma were also classified according to gender ((c) and (d)). Data are presented as means ± SEM. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001 versus the control group. Correlation analysis between miR-21 relative expression and AOPP levels (e), calculated using analysis of the Spearman correlation coefficient. C: control group; R: robust group; F: fragile group.

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Source: PubMed

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