摘要
目的 研究肺炎链球菌是否可触发肺Ⅱ型上皮细胞(A549)微丝肌动蛋白F-actin重排,进而导致对A549细胞的侵袭及其与磷脂酶C信号转导分子的关系。方法 采用F-actin特异性FITC-phalloidin荧光染料,观察肺炎链球菌作用A549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A549细胞,观察肺炎链球菌对A549细胞的侵袭数;使用PLC信号转导分子抑制剂U73122预处理A549细胞,观察其与F-actin细胞骨架重排百分率间是否存在剂量依赖关系。结果 肺炎链球菌作用A549细胞后,经FITC-phalloidin荧光染色,F-actin细胞骨架呈亮黄色圆形聚集;F-actin细胞骨架重排抑制剂细胞松弛素D可明显降低肺炎链球菌对A549细胞的侵袭,在其浓度为0.25μg/ml时,未得到可测的细菌数;PLC信号转导分子抑制剂U73122可部分抑制A549细胞F-actin细胞骨架重排,并与F-actin细胞骨架重排百分率间存在量效关系,其相关系数为rPLC=-0.98(P<0.05)。结论 肺炎链球菌可通过激活A549细胞PLC信号转导分子而触发A549细胞F-actin细胞骨架重排,而侵袭A549细胞。
Objective To investigate whether S. pn can provoke filamentous actin (F-actin) rearrangements in vitro , which will further lead to S. pn invasion of A549 and the relationship between PLC signaling molecule and. the invasion events. Methods Labelled F-actin with FITC-phalloidin, we observed F-actin rearrangements by S. pn adhesion of type Ⅱ pneumocytes ( A549). S. pn invasion of A549 cells was determined by pretreating A549 cells with Cytochalasin D . To investigate whether F-actin rearrangements can be blocked by PLC inhibitor, A549 cells were pretreated with PLC inhibitors U73122. Results Intact S. pn can promote F-actin rearrangements. Cytochalasin D is able to prevent S. pn invasion of A549 cells. Inhibitors of PLC signal transduction molecules block F-actin rearrangements dose dependently. Conclusion S. pn can provoke F-actin rearrangements through PLC signaling pathways, which will further lead to S. pn invasion of A549 cells.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2004年第7期456-459,共4页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金资助项目(30170050)
