盐皮质激素受体阻断剂对SiO_2诱导的肺巨噬细胞表型偏移的调节

Regulatory effects of spirolactone on SiO_2-induced lung macrophage subtype switching

目的:研究盐皮质受体阻断剂螺内酯(spirolactone,SPI)对二氧化硅(SiO_2)诱导的肺巨噬细胞表型偏移的影响。方法:将60只C57BL/6小鼠随机分为生理盐水(NS)组、SiO_2组、SPI治疗组(SiO_2+SPI组)及溶剂对照组(SiO_2+NS组),经口咽吸入SiO_2悬浊液(40 mg/kg)建立矽肺模型,对照组给予等量生理盐水,SiO_2+SPI组在造模后每天经口灌胃给予SPI(10 mg·kg^(-1)·d^(-1)),SiO_2+NS组以相同方式给予等量生理盐水。造模后3 d、14d、28 d取材,进行肺泡灌洗,取灌洗液细胞进行定量、分类计数及流式细胞术分析,右下肺叶用酶解法制成单细胞悬液行流式细胞术分析。结果:与SiO_2组相比,SPI治疗可以明显降低SiO_2干预后3 d、14 d支气管肺泡灌洗液总细胞数及巨噬细胞、中性粒细胞及淋巴细胞数。SiO_2干预后14 d、28 d,肺泡巨噬细胞和间质巨噬细胞由M1表型向M2表型偏移,SPI治疗可以有效逆转SiO_2诱导的巨噬细胞表型偏移。结论:SPI可以减轻SiO_2导致的肺泡炎症细胞浸润,可能是通过逆转SiO_2诱导的巨噬细胞表型偏移实现的。

AIM： To investigate the influence of spirolactone（ SPI） on mouse pulmonary macrophage subtype switching induced by silicon dioxide（ SiO2）. METHODS： C57BL/6 mice were divided into normal saline（ NS） group,SiO2 group,SiO2＋ SPI group and SiO2＋ NS group. A mouse model of silicosis was developed with crystalline SiO2 particles（ 40 mg / kg,via oropharyngeal instillation）. SPI（ 20 mg / kg） or（ NS） was delivered daily by oral gavage after SiO2 administration. The mice were sacrificed on days 3,14 and 28. Alveolar washing,total cell counting,differential cell counting and flow cytometry analysis were performed. The right lower lobe of lavaged lungs was collected to prepare singlecell suspension for flow cytometry analysis. RESULTS： Compared with SiO2 group,SPI treatment significantly alleviated SiO2-induced inflammatory cell accumulation in brochoalveolar lavage fluid on day 3 and 14. Compared with NS group,the M1 alveolar macrophages and M1 pulmonary interstitial macrophages in SiO2 group switched to M2 subtype dramatically,while SPI treatment reversed the switching effectively. CONCLUSION： SPI treatment alleviates SiO2-induced inflammatory cell accumulation,which may be related to reversing macrophage subtype switching.