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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 effects of SiO2 on the expression of advanced glycation end-product receptor (RAGE), ligand-calcitonin (S100) and high-mobility group histone-1 (HMGB1) The early prevention and treatment of silicosis provide a theoretical basis. Methods The bronchoalveolar macrophages of SD rats were collected and purified by bronchoalveolar lavage. The cells were counted as 1 × 106 cells / ml, divided into blank control group and experimental group. 100 μg / ml SiO2 was added to the experimental group and serum-free culture Liquid, cultured 4,24,48 h. The expressions of RAGE, S100 and HMGB1 in alveolar macrophages were detected by fluorescence immunocytochemistry, enzyme-linked immunosorbent assay and Western blot. The cultured fibroblasts were cultured for 4, 24, and 48 h in SiO2 experimental group and blank group, respectively. The proliferation of fibroblasts was measured by MTT assay. Results SiO2 could induce the high expression of S100 and HMGB1 in a time-dependent manner (P <0.05), but lower the expression of RAGE in the cell membrane than in the blank control group (P <0.05) The supernatant of SiO2 supernatant could induce the proliferation of lung fibroblasts as well as the time-effect relationship (P <0.05). Conclusion RAGE is highly expressed on the membrane of normal alveolar macrophages. SiO2 can reduce its expression. SiO2 can induce alveolar macrophages to secrete S100, HMGB1, and with the prolongation of time, increased secretion. SiO2 supernatant of AM can induce the proliferation of lung fibroblasts, and with the extension of time, the number increased.