Scanu A, Oliviero F, Luisetto R, Sfriso P, Dayer J M, Burger D, Punzi L
Cattedra e U.O.C. di Reumatologia, Università di Padova, Padova, Italia.
Reumatismo. 2010 Oct-Dec;62(4):266-72. doi: 10.4081/reumatismo.2010.266.
To evaluate whether high density lipoproteins (HDL) affect monosodium urate (MSU) crystal-induced inflammation in the murine air pouch model.
MSU crystals were prepared by Denko's method and sterilized by heating at 180°C for 2 h before each experiment. Human HDL were isolated from peripheral blood of healthy volunteers. MSU crystals (2 mg in 1 ml of PBS) were injected into subcutaneous air pouches in mice in the presence or absence of HDL (0.1 mg). Negative control pouches received 1 ml of PBS. To recover pouch fluid, the pouches were washed with 2 ml of PBS after the animals were sacrificed. The leukocyte count in the lavage fluids was obtained using a hemocytometer and differential leukocyte count was determined by May-Grunwald-Giemsa staining. IL-6, KC, CCL2 and TNF-α levels were measured in exudates by ELISA.
MSU crystals increased the number of leukocytes and the neutrophil migration, as well as the concentrations of IL-6, KC and CCL2 in pouch fluids, while the TNF-α levels were not detectable. The treatment with HDL led to a reduction in all inflammatory parameters: the leukocyte count decreased by 73%; the neutrophil density decreased by 35%; the IL-6, KC and CCL2 concentration decreased by 4-, 6- and 5-fold respectively.
This study shows that HDL may limit the inflammatory process by inhibiting leukocyte recruitment and cytokine release. HDL are likely to represent a mechanism of control of crystal-induced inflammation.
在小鼠气袋模型中评估高密度脂蛋白（HDL）是否影响尿酸单钠（MSU）晶体诱导的炎症。
采用Denko法制备MSU晶体，每次实验前在180°C加热2小时进行灭菌。从健康志愿者的外周血中分离人HDL。在有或无HDL（0.1mg）存在的情况下，将MSU晶体（2mg溶于1ml PBS）注射到小鼠皮下气袋中。阴性对照气袋注射1ml PBS。动物处死后，用2ml PBS冲洗气袋以回收袋内液体。使用血细胞计数器获得灌洗液中的白细胞计数，并通过May-Grunwald-Giemsa染色确定白细胞分类计数。通过ELISA测定渗出液中IL-6、KC、CCL2和TNF-α的水平。
MSU晶体增加了白细胞数量和中性粒细胞迁移，以及袋内液体中IL-6、KC和CCL2的浓度，而未检测到TNF-α水平。HDL治疗导致所有炎症参数降低：白细胞计数降低73%；中性粒细胞密度降低35%；IL-6、KC和CCL2浓度分别降低4倍、6倍和5倍。
本研究表明，HDL可能通过抑制白细胞募集和细胞因子释放来限制炎症过程。HDL可能代表了一种控制晶体诱导炎症的机制。
