Korean J Physiol Pharmacol.  2020 Sep;24(5):395-402. 10.4196/kjpp.2020.24.5.395.

Effects of troxerutin on vascular inflammatory mediators andexpression of microRNA-146a/NF-B signaling pathway in aortaof healthy and diabetic rats

• Xing C
• Xiang D
• Caiying L

• KMID: 2505774
• DOI: http://doi.org/10.4196/kjpp.2020.24.5.395

Abstract

This study has investigated the effect of a potent bioflavonoid, troxerutin,on diabetes-induced changes in pro-inflammatory mediators and expression ofmicroRNA-146a and nuclear factor-kappa-B (NF-κB) signaling pathway in aortic tissueof type-I diabetic rats. Male Wistar rats were randomly divided into four groups(n = 6/each): healthy, healthy-troxerutin, diabetic, and diabetic-troxerutin. Diabeteswas induced by streptozotocin injection (60 mg/kg; intraperitoneally) and lasted 10weeks. Troxerutin (150 mg/kg/day) was administered orally for last month of experiment.Inflammatory cytokines IL-1, IL-6, and TNF-, as well as intercellular adhesionmolecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), cyclooxygenase-II(COX-II), and inducible-nitric oxide synthase (iNOS) were measured on aortic samplesby enzyme-linked immunosorbent assay. Gene expressions for transcription factorNF-κB, interleukin-1 receptor-associated kinase-1 (IRAK-1), TNF receptor-associatedfactor-6 (TRAF-6), and microRNA-146a were determined using real-time polymerasechain reaction. Ten-week diabetes significantly increased mRNA levels of IRAK-1,TRAF-6, NF-κB, and protein levels of cytokines IL-1, IL-6, TNF-, adhesion moleculesICAM-1, VCAM, and iNOS, COX-II, and decreased expression of microRNA-146a ascompared with healthy rats (p < 0.05 to p < 0.01). However, one month treatmentof diabetic rats with troxerutin restored glucose and insulin levels, significantly decreasedexpression of inflammatory genes and pro-inflammatory mediators andincreased microRNA level in comparison to diabetic group (p < 0.05 to p < 0.01). Inhealthy rats, troxerutin had significant reducing effect only on NF-κB, TNF- and COXIIlevels (p < 0.05). Beside slight improvement of hyperglycemia, troxerutin preventedthe activation of NF-κB-dependent inflammatory signaling in

Keyword

Aorta; Diabetes mellitus; Nuclear factor-kappa B; Troxerutin; Vacscular injury

Figure

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  Fig. 1 Chemical structure of troxerutin.

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  Fig. 2 Real-time PCR analysis of genes expressions in the aortic tissue. The expression (mRNA) levels of NF-κB (A), IRAK-1 (B) and TRAF-6 (C). The data were expressed as mean ± standard error. n = 6 for each group. NF-κB, nuclear factor kappa B; IRAK-1, interleukin-1 receptor-associated kinase-1; TRAF-6, tumor necrosis factor receptor-associated factor-6; Cont, control; TXR, troxerutin. *p < 0.05, and **p < 0.01 vs. Healthy-Cont group; and #p < 0.05 vs. Diabetic-Cont group.

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  Fig. 3 Real-time PCR analysis of miR-146a expression in the aortic tissue. The data were expressed as mean ± standard error. n = 6 for each group. Cont, control; TXR, troxerutin. **p < 0.01 vs. Healthy-Cont group; and #p < 0.05 vs. Diabetic-Cont group.

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  Fig. 4 The levels of inflammatory cytokines in the endothelial cells of aortic tissue. IL-6 (A), IL-1β (B) and TNF-α (C). The data were expressed as mean ± standard error. n = 6 for each group. IL, interleukin; TNF-α, tumor necrosis factor-α; Cont, control; TXR, troxerutin. *p < 0.05 and **p < 0.01 vs. Healthy-Cont group; and #p < 0.05 vs. Diabetic-Cont group.

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  Fig. 5 The levels of adhesion molecules in aortic tissue. ICAM-1 (A), VCAM (B). The data were expressed as mean ± standard error. n = 6 for each group. ICAM-1, intercellular adhesion molecule-1; VCAM, vascular cell adhesion molecule; Cont, control; TXR, troxerutin. *p < 0.05 and **p < 0.01 vs. Healthy-Cont group; and #p < 0.05 vs. Diabetic-Cont group.

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  Fig. 6 The levels of inflammatory inducible enzymes in aortic tissue. iNOS (A), and COX-II (B). The data were expressed as mean ± standard error. n = 6 for each group. iNOS, inducible-nitric oxide synthase; COX-II, cyclooxygenase-II; Cont, control; TXR, troxerutin. *p < 0.05 and **p < 0.01 vs. Healthy-Cont group; and #p < 0.05 vs. Diabetic-Cont group.

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  Fig. 7 Conceptual diagram of the study. NF-κB, nuclear factor kappa B; IRAK-1, interleukin-1 receptor-associated kinase-1; TRAF-6, tumor necrosis factor receptor-associated factor-6; ICAM-1, intercellular adhesion molecule-1; IL, interleukin; TNF-α, tumor necrosis factor-α.

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