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目的 探索矽肺发病过程中 ,SiO2 对肺泡上皮细胞增殖和间隙连接通讯功能 (GJIC)的影响。方法 不同剂量的SiO2 刺激SD大鼠肺泡巨噬细胞 (PAM)培养上清液 ,作用于正常貂肺泡上皮细胞株CCL 6 4细胞。用四唑盐 (MTT)法测定CCL 6 4细胞增殖 (以A570nm值表示 ) ;采用激光扫描共焦显微镜 (LSCM ,LeicaTCSSP)用荧光光漂白后再恢复 (FRAP)技术测定CCL 6 4细胞GJIC功能 [以荧光扩散率K(× 10 -3 /s) ]表示。结果 体积分数为 5 %的SiO2 刺激的PAM上清液能诱导CCL 6 4细胞增殖 (F =9.6 79,P <0 .0 1) ,并抑制CCL 6 4细胞间GJIC功能 (F =2 0 .5 87,P <0 .0 1)。在 5 0~ 5 0 0 μg/mlSiO2 范围内 ,随SiO2 浓度增加 ,两者均呈良好的剂量依赖性关系 (GJIC :r =- 0 .943,P <0 .0 5 ;增殖 :r=0 .891,P <0 .0 5 )。结论 SiO2 刺激PAM培养上清液可以抑制肺泡上皮细胞的GJIC功能 ,并诱导细胞增殖。推论肺泡上皮细胞GJIC功能下调在SiO2 介导的肺泡上皮组织损伤中有重要的作用。
Objective To explore the effect of SiO2 on the proliferation and GJIC of alveolar epithelial cells in the pathogenesis of silicosis. Methods Different dosages of SiO2 were used to stimulate supernatant of alveolar macrophage (PAM) in SD rats, and the effect was on normal mink alveolar epithelial cell line CCL 6 4. The proliferation of CCL 6 4 cells was measured by MTT assay (A570 nm). The GJIC function of CCL 6 4 cells was determined by fluorescence photobleaching and restoring (FRAP) using laser scanning confocal microscope (LSCM, Leica TCSSP) [Expressed as fluorescence diffusivity K (× 10 -3 / s)]. Results The proliferation of CCL 6 4 cells induced by SiO 2 stimulated by 5% (F = 9.679, P <0.01) and inhibited the GJIC function of CCL 6 4 cells (F = 20. 5 87, P <0 01). In the range of 5 0 ~ 500 μg / ml SiO 2, the concentration of SiO 2 increased in a dose-dependent manner (GJIC: r = - 0.943, P <0.05; .891, P <0. 05). Conclusion SiO2 stimulated PAM culture supernatant can inhibit GJIC function of alveolar epithelial cells and induce cell proliferation. It is concluded that the down-regulation of GJIC function in alveolar epithelial cells plays an important role in SiO2-mediated alveolar epithelial injury.