Herrera Bruno S, Hasturk Hatice, Kantarci Alpdogan, Freire Marcelo O, Nguyen Olivia, Kansal Shevali, Van Dyke Thomas E
Department of Applied Oral Sciences, Forsyth Institute, Cambridge, Massachusetts, USA.
Department of Applied Oral Sciences, Forsyth Institute, Cambridge, Massachusetts, USA
Infect Immun. 2015 Feb;83(2):792-801. doi: 10.1128/IAI.02444-14. Epub 2014 Dec 8.
Diabetic complications involve inflammation-mediated microvascular and macrovascular damage, disruption of lipid metabolism, glycosylation of proteins, and abnormalities of neutrophil-mediated events. Resolution of inflamed tissues to health and homeostasis is an active process mediated by endogenous lipid agonists, including lipoxins and resolvins. This proresolution system appears to be compromised in type 2 diabetes (T2D). The goal of this study was to investigate unresolved inflammation in T2D. Wild-type (WT) and genetically engineered mice, including T2D mice (db/db), transgenic mice overexpressing the human resolvin E1 (RvE1) receptor (ERV1), and a newly bred strain of db/ERV1 mice, were used to determine the impact of RvE1 on the phagocytosis of Porphyromonas gingivalis in T2D. Neutrophils were isolated and incubated with fluorescein isothiocyanate-labeled P. gingivalis, and phagocytosis was measured in a fluorochrome-based assay by flow cytometry. Mitogen-activated protein kinase (MAPK) (p42 and p44) and Akt (Thr308 and Ser473) phosphorylation was analyzed by Western blotting. The mouse dorsal air pouch model was used to evaluate the in vivo impact of RvE1. Results revealed that RvE1 increased the neutrophil phagocytosis of P. gingivalis in WT animals but had no impact in db/db animals. In ERV1-transgenic and ERV1-transgenic diabetic mice, phagocytosis was significantly increased. RvE1 decreased Akt and MAPK phosphorylation in the transgenic animals. In vivo dorsal air pouch studies revealed that RvE1 decreases neutrophil influx into the pouch and increases neutrophil phagocytosis of P. gingivalis in the transgenic animals; cutaneous fat deposition was reduced, as was macrophage infiltration. The results suggest that RvE1 rescues impaired neutrophil phagocytosis in obese T2D mice overexpressing ERV1.
糖尿病并发症涉及炎症介导的微血管和大血管损伤、脂质代谢紊乱、蛋白质糖基化以及中性粒细胞介导事件的异常。炎症组织恢复至健康和内稳态是一个由内源性脂质激动剂介导的活跃过程，这些激动剂包括脂氧素和消退素。在2型糖尿病（T2D）中，这种促消退系统似乎受到损害。本研究的目的是调查T2D中未消退的炎症。使用野生型（WT）和基因工程小鼠，包括T2D小鼠（db/db）、过表达人消退素E1（RvE1）受体（ERV1）的转基因小鼠以及新培育的db/ERV1小鼠品系，来确定RvE1对T2D中牙龈卟啉单胞菌吞噬作用的影响。分离中性粒细胞并与异硫氰酸荧光素标记的牙龈卟啉单胞菌一起孵育，通过基于荧光染料的流式细胞术测定吞噬作用。通过蛋白质免疫印迹法分析丝裂原活化蛋白激酶（MAPK）（p42和p44）以及Akt（Thr308和Ser473）的磷酸化情况。使用小鼠背部气囊模型评估RvE1的体内影响。结果显示，RvE1增加了WT动物中牙龈卟啉单胞菌的中性粒细胞吞噬作用，但对db/db动物没有影响。在ERV1转基因和ERV1转基因糖尿病小鼠中，吞噬作用显著增加。RvE1降低了转基因动物中Akt和MAPK的磷酸化。体内背部气囊研究表明，RvE1减少了转基因动物中中性粒细胞向气囊的流入，并增加了牙龈卟啉单胞菌的中性粒细胞吞噬作用；皮肤脂肪沉积减少，巨噬细胞浸润也减少。结果表明，RvE1可挽救过表达ERV1的肥胖T2D小鼠中受损的中性粒细胞吞噬作用。
