[晚期糖基化终产物受体在脂多糖诱导的小鼠肺微血管内皮细胞细胞骨架变化中的作用]
[Role of RAGE in lipopolysaccharide-induced cytoskeletal changes in mouse pulmonary microvascular endothelial cells].
作者信息
Zhou Xiao-Yan, Zhang Wei-Jin, Huang Qiao-Bing, Guo Xiao-Hua
机构信息
Department of Pathophysiology of Southern Medical University, Key Laboratory of Shock and Microcirculation of Guangdong Province, Guangzhou 510515, China.E-mail:
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2015 Jan;35(1):6-11.
OBJECTIVE
To investigate lipopolysaccharide (LPS)-induced changes of cytoskeletal filamentous actin in primary isolated pulmonary microvascular endothelial cells (PMVECs) from wild-type and RAGE knock-out mouse.
METHODS
The lungs of wild-type and RAGE knock-out mice were digested with collagenase type I to obtain endothelial cells purified by anti-CD31-coupled magnetic beads. The PMVEC identified by factor VIII labeling were stimulated with LPS at different concentrations and the changes of filamentous actin were observed by confocal microscopy.
RESULTS
The cultured primary cells showed typical endothelial cell phenotype as examined with factor VIII labeling. LPS stimulation caused rearrangement of the cytoskeletal filament F-actin in wild-type mouse PMVECs with stress fiber formation, but such changes were not obvious in RAGE knock-out mouse PMVECs.
CONCLUSION
Mouse PMVECs of a high purity can be obtained by immune magnetic beads. RAGE is involved in LPS-induced destruction of mouse PMVEC cytoskeletons.
目的
研究脂多糖（LPS）诱导的野生型和RAGE基因敲除小鼠原代分离肺微血管内皮细胞（PMVECs）中细胞骨架丝状肌动蛋白的变化。
方法
用I型胶原酶消化野生型和RAGE基因敲除小鼠的肺，以获得通过抗CD31偶联磁珠纯化的内皮细胞。用不同浓度的LPS刺激经VIII因子标记鉴定的PMVECs，通过共聚焦显微镜观察丝状肌动蛋白的变化。
结果
培养的原代细胞经VIII因子标记检测显示典型的内皮细胞表型。LPS刺激导致野生型小鼠PMVECs中细胞骨架丝状肌动蛋白F-肌动蛋白重排并形成应力纤维，但在RAGE基因敲除小鼠PMVECs中这种变化不明显。
结论
通过免疫磁珠可获得高纯度的小鼠PMVECs。RAGE参与LPS诱导的小鼠PMVEC细胞骨架破坏。
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