趋化因子受体2抑制剂在急性痛风小鼠模型中的作用研究

Role of chemokine receptor 2 inhibitor in mouse model of acute gout

目的探讨趋化因子受体2(CXCR2)抑制剂在急性痛风小鼠模型中的作用。方法小鼠急性痛风足掌模型:12只6~8周龄C57BL/6雄性小鼠随机分为3组,每组4只,对照组:小鼠右后足掌注射生理盐水;实验组:小鼠右后足掌注射单钠尿酸盐(MSU)建立急性痛风足掌模型;干预组:小鼠右后足掌注射MSU及CXCR2抑制剂SB225002。于注射8 h后测量3组小鼠右/左后足掌厚度比值,采用苏木素-伊红(HE)染色评估小鼠足掌关节炎症程度,采用蛋白质免疫印迹法(Western blot)检测足掌组织白细胞介素(IL)-1β水平。小鼠急性痛风腹腔模型:9只6~8周龄C57BL/6雄性小鼠随机分为3组,每组3只。对照组:小鼠腹腔注射生理盐水;实验组:小鼠腹腔注射MSU建立急性痛风腹腔模型;实验组:小鼠腹腔注射MSU及CXCR2抑制剂SB225002。于MSU注射4 h后收取腹腔灌洗液检测中性粒细胞密度及比例、IL-1β及趋化因子水平。结果在小鼠急性痛风足掌模型中,实验组小鼠右/左后足掌厚度比值高于对照组,干预组低于实验组(P<0.05);与对照组比较,实验组小鼠足掌关节炎症细胞浸润程度明显增加;与实验组比较,干预组小鼠足掌关节炎症细胞浸润程度明显下降;实验组足掌组织IL-1β水平高于对照组,干预组低于实验组。在小鼠急性痛风腹腔模型中,实验组小鼠腹腔灌洗液中中性粒细胞密度和比例均高于对照组,干预组上述指标均低于实验组(P<0.001或P<0.05);实验组小鼠腹腔灌洗液中IL-1β、CXCL1、CXCL2、CXCL8水平均高于对照组,干预组小鼠腹腔灌洗液中CXCL1、CXCL2、CXCL8水平均低于实验组(P<0.05)。结论抑制CXCR2可减轻小鼠急性痛风炎症反应,CXCR2可能成为急性痛风治疗中抑制急性炎症的可行靶点。

Objective To explore the role of chemokine receptor 2(CXCR2)inhibitor in mouse model of acute gout.Methods Pawls model of acute gout mice:12 C57 BL/6 male mice aged 6 to 8 weeks old were randomly divided into 3 groups(4 mice in each group).Mice in control group were injected with saline in the right hind pawl.Mice in experimental group were injected with MSU in the right hind pawl to establish pawls model of acute gout.Mice in intervention group were injected with MSU and CXCR2 inhibitor SB225002 in the right hind pawl.Right/left hind paw thickness ratio of mice in 3 groups was measured 8 h after injection.Degree of mice paw joint inflammation was assessed by hematoxylin-eosin(HE)staining.Interleukin(IL)-1 level was measured by western blotting.Peritoneal model of acute gout mice:9 C57 BL/6 male mice aged 6 to 8 weeks old were randomly divided into 3 groups(3 mice in each group):Mice in control group were injected with saline intraperitoneally,mice in experimental group were injected with MSU intraperitoneally to establish peritoneal model of acute gout,mice in intervention group were injected with MSU and CXCR2 inhibitor SB225002 intraperitoneally.Intraperitoneal lavage fluid was collected for detection of neutrophil density and proportion,IL-1 and chemokines levels 4 h after MSU injection.Results In pawls model of acute gout mice,right/left hind paw thickness ratio in experimental group was higher than that in control group,and the ratio in intervention group was lower than that in experimental group(P<0.05);compared with the control group,infiltration degree of inflammatory cell in pawl was significantly increased in experimental group,while compared with experimental group,it was significantly reduced in intervention group;IL-1 level of paw tissue was higher in experimental group than that in control group,and the intervention group was lower than that in experimental group.In peritoneal model of acute gout mice,density and proportion of neutrophil of peritoneal lavage fluid in experimental group were higher than those in control group,and above indexes in intervention group were lower than those in experimental group(P<0.001 or P<0.05);levels of IL-1,CXCL1,CXCL2 and CXCL8 in experimental group were significantly higher than those in control group,and levels of CXCL1,CXCL2 and CXCL8 were lower in intervention group than those in experimental group(P<0.05).Conclusion Inhibition of CXCR2 can reduce acute inflammatory response in mouse model of acute gout.CXCR2 may be a viable target to inhibit acute inflammation in the treatment of acute gout.