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[目的]探讨SiO2是否通过影响矽肺患者肺泡巨噬细胞(AM)和肺成纤维细胞(FB)基质金属蛋白酶(MMPs)/金属蛋白酶组织抑制因子(TIMPs)系统参与矽肺纤维化的发生发展。[方法]收集矽肺患者的AM,在体外以SiO2(50μg/ml)和不加血清的DMEM培养基培养2、6、12、18、24、36h,用免疫细胞化学的方法检测AM中MMP-9和TIMP-1的表达。收集矽肺患者AM培养上清,与人胚肺FB共同孵育6、12、18、24、36、48h,用免疫细胞化学的方法检测FB中MMP-1和TIMP-1的表达。[结果]经SiO2刺激的AMMMP-9表达明显上调,且随刺激时间不同表达量也不同,于18h达高峰(平均光密度值为0.440±0.021),与同期对照组AM(0.390±0.011)相比,差异有统计学意义(P﹤0.05);而TIMP-1的表达与对照组(平均光密度值分别为0.175±0.019、0.162±0.044)的差异无统计学意义(P﹥0.05)。未经SiO2刺激的矽肺患者AM培养上清作用于FB24h,FB中MMP-1的表达较空白对照组减少(平均光密度值分别为0.103±0.014、0.133±0.023,P﹤0.01),TIMP-1的表达增多(平均光密度值分别为0.108±0.012、0.065±0.006,P﹤0.01);经SiO2刺激24h的矽肺患者AM培养上清作用于FB后,FB中MMP-1的表达量进一步减少(平均光密度值分别为0.062±0.008、0.133±0.023,P﹤0.01),而TIMP-1的表达量进一步增加(平均光密度值分别为0.143±0.015、0.065±0.006,P﹤0.01)。[结论]SiO2上调AM中MMP-9的表达,可能与肺脏损伤早期,降解基底膜,导致包括成纤维细胞在内的多种细胞迁移有关。通过肺泡巨噬细胞的介导,SiO2影响了FB中MMP/TIMP系统的平衡,参与了矽肺纤维化的发生发展。
[Objective] To explore whether SiO 2 participates in the occurrence and development of silicosis through affecting the alveolar macrophages (AMs) and fibroblasts (FBs) of matrix metalloproteinases (MMPs) / tissue inhibitors of metalloproteinases (TIMPs) in silicosis patients. [Methods] The AM of silicosis patients were collected. The cells were cultured for 2, 6, 12, 18, 24, 36 h in vitro with SiO2 (50μg / ml) and serum-free DMEM medium. 9 and TIMP-1 expression. AM supernatant from patients with silicosis was collected and incubated with human embryonic lung FB for 6, 12, 18, 24, 36 and 48 hours. The expression of MMP-1 and TIMP-1 in FB was detected by immunocytochemistry. [Results] The expression of AMMMP-9 stimulated by SiO2 was significantly up-regulated, and the expression of AMMMP-9 was also different with different stimulation time. The peak reached the peak at 18h (mean optical density was 0.440 ± 0.021) (P <0.05). There was no significant difference between the expression of TIMP-1 and the control group (mean optical density values were 0.175 ± 0.019,0.162 ± 0.044 respectively) (P> 0.05). The supernatant of AM in non-SiO2-stimulated silicosis patients was treated with FB24h and the expression of MMP-1 in FB was decreased compared with the control group (mean optical density was 0.103 ± 0.014,0.133 ± 0.023, P <0.01) (Mean optical density 0.108 ± 0.012, 0.065 ± 0.006 respectively, P <0.01). After AMI supernatant was stimulated by SiO2 for 24 h, the expression of MMP-1 in FB was further decreased The average optical density values were 0.062 ± 0.008,0.133 ± 0.023, P <0.01), while the expression of TIMP-1 was further increased (average optical density values were 0.143 ± 0.015,0.065 ± 0.006, P <0.01). [Conclusion] SiO2 up-regulates the expression of MMP-9 in AM, which may be related to the early stage of lung injury, the degradation of basement membrane, leading to the migration of many kinds of cells including fibroblasts. Through the mediation of alveolar macrophages, SiO2 affects the balance of the MMP / TIMP system in FB and participates in the occurrence and development of silicosis fibrosis.