矽肺肺泡巨噬细胞对肺成纤维细胞基质金属蛋白酶及其抑制因子和胶原表达的影响

Effect of human silicotic alveolar macrophages on expression of matrix metaHoproteinase, tissue inhibitor of metalloproteinase and collagen in human lung fibroblasts

目的探讨经SiO：刺激的肺泡巨噬细胞（AM）通过人胚肺成纤维细胞（HELF）对基质金属蛋白酶-1（MMP-1）及其抑制因子（TIMP-1）和Ⅲ型胶原表达的影响。方法收集矽肺患者AM，将其分为加入SiO2粉尘悬液的处理组和仅加入无血清培养液的对照组。培养18h后吸出培养上清。将原代培养的HELF分为4组：（1）处理组：加入经SiO2刺激的AM上清；（2）对照组：加入未经SiO2刺激的AM上清；（3）10％血清组：仅加入含体积分数为10％胎牛血清的DMEM；（4）空白对照组：仅加入含体积分数为3％胎牛血清的DMEM。4组均分别于6、12、18、24、36、48h取出细胞爬片，吸出上清，用免疫细胞化学方法检测HELF中MMP-1和TIMP-1的表达，用Western blot法检测HELF条件培养上清中Ⅲ型胶原的表达。结果处理组18、24、36、48hMMP-1表达分别为0．0605±0．0201，0．0519±0．0117，0．0412±0．0105，0．0213±0．0106，较对照组和空白对照组明显降低，差异均有统计学意义（P〈0．05，P〈0．01）。与对照组和空白对照组相比，处理组各时间点TIMP一1表达和Ⅲ型胶原含量明显增高，差异均有统计学意义（P〈0．05，P〈O．01），TIMP-1／MMP-1与Ⅲ型胶原呈正相关（r=0．88，19〈0．01）。结论SiO2经AM的介导可促进FB表达TIMP-1和Ⅲ型胶原，抑制HELF表达TIMP-1，TIMP-1／MMP-1的失衡与Ⅲ型胶原的病理性累积有关。

Objective To study the effect of culture supernatant of alveolar macrophage alveolar macrophages（AM） stimulated by SiO2 on the expression of matrix metalloproteinases（MMP-1）,tissue inhibitor of metalloproteinase-1 （TIMP-1） and collagen of fibroblast human embroyonic lung fibroblasts （HELF） in the development of silicosis fibrosis. Methods AMs were collected from a silicotic patient by bronchoalveolar lavage and exposed to SiO2, cultured human embryo lung fibroblast were allocated into a treated group, a control group, a positive group, and a blank group. HELF was incubated with the cultured supernatant of AMs for 6, 12, 18, 24, 36, 48 h. Immunocytochemical and Western blot technology were used to detect MMP-1 and TIMP-1 expressions in HELF and collagen expression in supernatant of HELF respectively. Results The supernatant of AM exposed to SiO2 significantly decreased the expressions of MMP-1 （0.0605±0.0201,0.0519± 0.0117, 0.0412±0.0105 and 0.0213±0.0106 in the treated group at 18, 24, 36 and 48 h） compared with the control group and the blank group （P〈0.05, P〈0.01 ） but stimulated expressions of TIMP-1 and collagen （P〈 0.05 ,P〈0.01 ）. The ratio of TIMP-1 to MMP-1 increased. The ratio of TIMP-1 to MMP-1 was positively correlated with the expression of collagen m （r=0.88, P〈0.01 ）. Conclusion Through AM mediation SiO2 can accelerate the expression of TIMP-1 and collagen, and inhibit the expression of MMP-1. The imbalance between the expression of TIMP-1 and that of MMP-1 is related with the abnormal increase in collagen Ⅲ.