[PubMed] [Google Scholar]Paolisso G, Rizzo MR, Mazziotti G, Tagliamonte MR, Gambardella A, Rotondi M, Carella C, Giugliano D, Varricchio M, D’Onofrio F. role of mitochondrial reactive oxygen species in regulation of the NF-B pathway and corresponding age-related inflammatory activation of endothelium. and AC710 [20-25]. In the current study, we used SkQ1 antioxidant, based on the plastoquinone moiety linked to dodecyltriphenylphosphonium cation that targets SkQ1 to mitochondria . SkQ1 and its analogs are efficient in the prevention of some age-associated pathologies, and they have therapeutic effects in animal models of diseases associated with inflammatory response (heart, brain, and kidney ischemic injury [27, 28], pyelonephritis , vision diseases [30, 31], sarcopenia , and dermal wound healing ). SkQ1 delays the development of various markers of aging and prolongs the lifespan of various animals [20, 22, 34, 35]. The antiinflammatory and vasoprotective action of SkQ1 could underlie some of these effects. Using mitochondria-targeted antioxidants, we show that mtROS are critical for the increase in CAM expression both in aortas of aged mice and in endothelial cells treated with TNF acting through the NF-B pathway. RESULTS Mitochondria-targeted antioxidant SkQ1 inhibits expression of adhesion molecules ICAM1 in the aortas of aged mice In aortal tissue of aged (24 month) CBAxC57bl/6 mice, mRNA expression of inflammatory markers such as adhesion molecules ICAM1 and VCAM and cytokines TNF and MCP1 were higher than in young (8-month-old) animals (Fig. ?(Fig.1).1). Long-term consumption of the mitochondria-targeted antioxidant SkQ1 (100 nmol/kg body weight per Rabbit Polyclonal to SLC25A6 day, 8 months) decreased mRNA expression of ICAM1 to the level of young animals (Fig. ?(Fig.1).1). SkQ1 also slightly decreased expression of other markers of inflammation, though these effects were not statistically significant. In aged mice the levels of the inflammatory cytokines TNF and IL-6 in the blood plasma were higher than in young animals, in agreement with previously published data [36-38]. SkQ1 treatment did not significantly influence the level of these cytokines in aged mice (Fig. ?(Fig.1).1). Thus, SkQ1 did not suppress the generation AC710 of TNF and IL-6, but it inhibited the activation effect of these cytokines in the aortic tissue. These data suggest that mtROS are involved in inflammatory response of endothelium in aged mice. Open in a separate window Physique 1 SkQ1 suppresses age-related increase in mRNA expression of some inflammatory markers in aortas of aged mice(A) ICAM1; (B) VCAM; (C) TNF; (D) MCP-1. The animals were treated as indicated on Fig. 1. Data are represented as mean +/? SEM. AC710 n = 10. ** p 0.001. Open up in another window Shape 2 Long-term SkQ1 treatment (250 nmol/kg each day, 8 weeks) will not influence circulatory degrees of proinflammatory cytokines in outdated (two years) versus youthful (six months) mice(A) TNF; (B) IL-6. Data are displayed as mean +/? SEM. n = 10. SkQ1 inhibits TNF-induced activation of endothelium To review the part of mtROS in the inflammatory response of endothelium, we looked into the result of SkQ1 on activation of endothelial cells in tradition activated with TNF. TNF can be widely used to review inflammatory response in both major cell cultures and in immortalized AC710 cell lines. Inside our function, we used the principal endothelial cell tradition HUVEC as well as the immortalized EA.hy926 cell line founded as the right model in lots of research [39, 40]. TNF-induced endothelial activation was evaluated using the next requirements: (i) upsurge in mRNA manifestation degree of adhesion substances ICAM, VCAM, and E-selectin; (ii) upsurge in ICAM1 manifestation on the top of cells; (iii) upsurge in IL-6 and IL-8 secretion; and (iv) upsurge in adhesion of human being promyelocytic leukemia cells (HL-60) towards the endothelial monolayer. SkQ1 suppresses TNF-induced mRNA manifestation of adhesion substances TNF (50 pg/ml) significantly increased mRNA manifestation of ICAM1 in both HUVEC and EA.hy926 endothelial cells (Fig. ?(Fig.3A,3A, ?,4A).4A). TNF considerably improved manifestation of E-selectin and VCAM in HUVEC also, however, not in EA.hy926 (Fig. ?(Fig.4B).4B). This difference between your cell lines was referred to previously . SkQ1 suppressed both fundamental and TNF-induced mRNA manifestation of ICAM1 (Fig. ?(Fig.3A,3A, ?,4A),4A), E-selectin, and VCAM (Fig. 4B, C). The result of SkQ1 was dose-dependent, and its own most effective focus were 0.2 nM. The traditional antioxidants N-acetylcysteine (NAC) (5 mM) and Trolox (0.1 mM) also reduced both fundamental and TNF-induced mRNA expression degree of ICAM1 (Fig. ?(Fig.3B),3B), related to published data [42-45] previously. It really is noteworthy how the SkQ1 analogs SkQBerb AC710 and SkQR1, holding berberine and rhodamine-19 cationic organizations, correspondingly , got the same actions (data not demonstrated). Open up in another window.