摘要
目的:空间失重条件下机体会出现心血管功能失调,血管内皮细胞的改变情况可能是导致此种异常的直接原因,本研究对模拟微重力(MMG)干预下人脐静脉内皮细胞(HUVECs)的黏着斑(FAs)多种参数进行半定量观测,并对FAs主要受体整合素(integrins)的表达进行评价,以期探索心血管功能失调的内在机制。方法:激光共聚焦结合高通量激光半定量计算方法,对FAs的各项参数进行评价,其参数包括FAs平均数量(VN);FAs平均面积(VA);FAs平均交叠面积(OSP);FAs平均到细胞边缘距离(DS)。Western blot以及RT-PCR检测integrinβ1和β4的表达。结果:FAs在MMG干预下,形成的数量(VN)与面积(VA)明显减少,VN较NG对照组减少-50.6%(P<0.05),VA较NG对照组减少-60.2%(P<0.05),FAs定位指标DS也不同于NG对照组-28.3%(P<0.05);MMG可导致integrinβ1和β4在mRNA与蛋白水平表达下调。结论:MMG可能通过影响HUVECs的FAs的生成与定位,以及下调integrins表达水平抑制内皮细胞增殖与迁移的;而FAs与integrins正是细胞进行胞内外增殖、迁移等信号交流的窗口,这也是导致产生心血管功能失调的直接原因之一。
Objective： The cardiovascular system undergoes major changes in stress with space flight; the endothelial cells may play a main role in this process. The objective of this study was to investigate this part to detect the parameters of FAs （mean vinculin spot number per cell, VN; mean vinculin spot area, VA; the area percentage of overlap of FAs OSP; average distance of vineulin spots from the cell edge, DS）; and to evaluate the expression of kinases and integrins of FAs. Methods： Semiquantitative Fas parameters was detected by confocal and high-pass filtered image. The expressions of integrins were detected by RT-PCR and Western blot. Result： After 48 h of microgravity exposure, the HUVECs presented a major reduction in VN （-50%） （P〈0.05） and VA（-60%） （P〈0.05）, and the DS were different from NG controls （-28%） （P〈0.05）; MMG inhibited the expression of integrinβ 1 and 13 4 both in protein and mRNA level. Conclusion： MMG could affect FAs formation and location, moreover, it could down-regulate integrins expression.
出处
《现代生物医学进展》
CAS
2013年第29期5625-5630,共6页
Progress in Modern Biomedicine
基金
国家自然科学基金青年科学基金项目(81102678)
关键词
模拟微重力MMG
脐静脉内皮细胞HUVECs
黏着斑
整合素
Modeled microgravity
MMG
Human umbilical vein endothelial cells
HUVECs
Focal adhesions, FAs
Integrin
