SiO_2刺激肺泡巨噬细胞对晚期糖基化终末产物受体及配体的影响  

Effect of stimulation to alveolar macrophage by SiO_2 on advanced glycation end product receptor and ligand

作　　者：白宇[1] 李素平[1] 付志华[2] 葛伦睿[3] 

机构地区：[1]山西医科大学公共卫生学院,山西太原030001 [2]太原市中心医院,山西太原030001 [3]复旦大学医院管理处,上海200032

出　　处：《中国工业医学杂志》2013年第2期92-94,157,共4页Chinese Journal of Industrial Medicine

摘　　要：目的探讨SiO2刺激肺泡巨噬细胞(AM)对晚期糖基化终末产物受体(RAGE)及配体——钙粒蛋白(S100)、高迁移率组蛋白-1(HMGB1)的影响,以为矽肺的早期防治提供理论依据。方法支气管灌洗法收集SD大鼠肺泡巨噬细胞进行纯化培养并计数细胞为1×106个/ml,分空白对照组和实验组,实验组加100μg/ml SiO2,空白对照组加无血清培养液,培养4、24、48h。用荧光免疫细胞化学方法、酶联免疫吸附法、Western blot检测肺泡巨噬细胞RAGE、S100和HMGB1的表达。将培养4、24、48 h的SiO2实验组及空白组AM培养上清液培养肺成纤维细胞24h,用MTT法测量成纤维细胞增殖情况。结果 SiO2能诱导细胞S100、HMGB1的高表达并有时间-效应关系(P<0.05),但会降低细胞膜RAGE表达,且低于空白对照组,亦具时间-效应关系(P<0.05);含SiO2的AM培养上清液能诱导肺成纤维细胞的增殖,也有时间-效应关系(P<0.05)。结论 RAGE在正常肺泡巨噬细胞膜上高表达;SiO2可以降低其表达。SiO2能诱导肺泡巨噬细胞分泌S100、HMGB1,且随作用时间的延长,分泌增多。SiO2的AM培养上清液能诱导肺成纤维细胞的增殖,且随作用时间的延长,数量增加。Objective To investigate the effect of stimulation to alveolar macrophage by SiOz on the receptor of advanced glycation end products （RAGE） and its ligands including calcium binding protein or calgranulin （S100）, high mobility group box-1 protein （HMGB1） thereby find a theoretical basis for the early prevention of silicosis. Methods SD rat alveolar macrophages were collected by bronchial lavage, through purification and cultivation, dilute these cells to the concentration of 1 × 10^6/ml, and divided into blank control group （with serum-free medium） and experimental group （with 100μg/ml of SiO2 ）, cultured for 4, 24, 48 hours. Then the expressions of RAGE, S100 and HMGB1 in alveolar macrophages were measured by immunofluorescence technic, enzyme-linked immunosorbent assay and Western blot analysis, respectively. Meanwhile, the culturing supernatant of both control group and experimental group were used to cultivate lung fibroblasts for 24 hours, then measure the proliferation situation of fibroblasts by MTI＇ assay. Results The results showed that SiOz could induce high-expression of S100 and HMGB1 of alveolar macrophages with time-effect relationship （ P 〈 0.05 ）, but decreased the expression of membrane RAGE （ even lower than that of blank control group）, which also showed time-effect relationship （P 〈 0. 05）. Addi tionally, the results showed that SiO2-contained AM culturing supernatant could induce proliferation of fibroblasts, which was time-effect relationship as well （ P 〈 0. 05 ）. Conclusion The results suggested that （ 1 ） RAGE is highly expressed on the membrane of normal alveolar macrophages, SiO2 may reduce its expression; （2） SiO2 can induce alveolar macrophages to secrete S100 and HMGB1, which has time-effect relationship; （3） SiO2 contained AM culturing supernatant may stimulate proliferation of fibroblasts, and the effect will be risen with culturing time.

关 键 词：晚期糖基化终末产物受体(RAGE) 配体 高迁移率组蛋白-1 钙粒蛋白 肺泡巨噬细胞 

分 类 号：R994[医药卫生—毒理学]