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目的 :通过体外实验 ,研究肺炎链球菌 (Streptococcus pneumoniae,S.pn)是否可触发肺 型上皮细胞 (A5 4 9)信号转导途径触发微丝肌动蛋白 (filam entous actin,F- actin)细胞骨架重排 ,进而侵袭 A5 4 9细胞 ,并初步分析触发 A5 4 9细胞 F- actin细胞骨架重排的细菌亚组分。方法 :采用 F- actin特异性 FITC-phalloidin荧光染料 ,观察 S.pn作用 A5 4 9细胞前后的 F- actin细胞骨架重排情况 ,依照重排百分率得分标准以 (% )表示 ;用 F- actin细胞骨架重排抑制剂细胞松弛素 D预处理 A5 4 9细胞 ,观察 S.pn对 A5 4 9细胞侵袭的改变情况 ;用变溶菌素提取 S.pn细胞壁以观察其对 F- actin细胞骨架重排的影响。结果 :S.pn作用A5 4 9细胞后 ,经 FITC- phalloidin荧光染色 ,F- actin细胞骨架呈黄绿色块状聚集 ,对照细胞呈现均匀黄绿色荧光外观 ;F- actin细胞骨架重排抑制剂细胞松弛素 D可明显降低 S.pn对 A5 4 9细胞的侵袭 ,在其浓度为 0 .2 5 μg/ m l时 ,未得到可测的细菌数 ;S.pn细胞壁作用 A5 4 9细胞后 ,经 FITC- phalloidn荧光染色 ,F-actin细胞骨架呈黄绿色块状聚集 ,二者存在剂量依赖性。结论 :S.pn及其细胞壁亚组分可触发 A5 4 9细胞F- actin细胞骨架重排 ,进而侵袭 A5 4 9细胞
OBJECTIVE: To investigate whether Streptococcus pneumoniae (S.pn) can trigger the signaling pathway of lung epithelial cells (A549) to trigger the filamentation of actin actin (F-actin) cells in vitro Skeleton rearrangement, and then invasion of A549 cells, and preliminary analysis of A549 cell trigger F-actin cytoskeleton rearrangement of the bacterial sub-components. Methods: F-actin-specific FITC-phalloidin fluorescent dye was used to observe the cytoskeletal rearrangement of F actin in A549 cells treated with S.pn. The percentages of rearrangement were expressed as% Cytoskeleton rearrangement inhibitor cytochalasin D pretreatment of A549 cells, to observe S.pn A549 cell invasion changes; with mutans bacilli extract S.pn cell wall to observe the F-actin cytoskeleton weight Row of impact. RESULTS: After A549 cells were treated with S.pn, the F-actin cytoskeleton was aggregated in yellow-green color with FITC-phalloidin staining, and the control cells showed a homogeneous yellow-green fluorescence appearance. F-actin cytoskeleton rearrangement inhibitor cells Relaxin D significantly decreased the invasion of A549 cells by S.pn, but no measurable bacterial counts were obtained at the concentration of 0.25 g / ml. After A549 cells were treated with S.pn cell wall, FITC-phalloidn fluorescence staining, F-actin cytoskeleton yellow-green mass aggregation, both in a dose-dependent manner. CONCLUSIONS: S.pn and its cell wall sub-components can trigger the cytoskeletal rearrangement of F-actin in A549 cells and further infect A549 cells