周细胞Toll样受体-4的脂多糖激活调节共培养通透性。
Lipopolysaccharide activation of pericyte's Toll-like receptor-4 regulates co-culture permeability.
作者信息
Edelman David A, Jiang Yang, Tyburski James G, Wilson Robert F, Steffes Christopher P
机构信息
Department of Surgery, Wayne State University, 3900 John R, Suite 400 POB, Detroit, MI 48201, USA.
出版信息
Am J Surg. 2007 Jun;193(6):730-5. doi: 10.1016/j.amjsurg.2006.08.086.
BACKGROUND
Pericytes (PCs) have a synergistic relationship with endothelial cells (MVEC) in regulating capillary permeability. PCs express Toll-like receptor-4 (TLR-4). We hypothesize one mechanism of MVEC/PC co-culture permeability is regulated through lipopolysaccharide (LPS) activation of pericyte TLR-4.
METHODS
Rat PCs were harvested and cultured. PCs were transfected with siRNA targeted to TLR-4. Western blotting was used to confirm gene silencing of TLR-4. A previously described co-culture permeability assay was performed after LPS treatment.
RESULTS
Western blot confirmed successful silencing of TLR-4 in PCs, which was sustained for 7 days. A dose- and time-dependent effect of LPS on albumin clearance was seen in MVEC/PC co-cultures. Co-cultures with TLR-4 silenced in PCs eliminated the LPS dose-dependent increase in albumin clearance.
CONCLUSIONS
TLR-4 regulates pericyte mediated capillary leak seen with LPS exposure. Our TLR-4 silencing model can be used to further investigate TLR-4's role in pericyte mediated capillary leak.
背景
周细胞（PCs）与内皮细胞（MVEC）在调节毛细血管通透性方面存在协同关系。周细胞表达Toll样受体4（TLR-4）。我们推测MVEC/PC共培养通透性的一种机制是通过脂多糖（LPS）激活周细胞TLR-4来调节的。
方法
收获并培养大鼠周细胞。用靶向TLR-4的小干扰RNA（siRNA）转染周细胞。采用蛋白质印迹法确认TLR-4基因沉默。在LPS处理后进行先前描述的共培养通透性测定。
结果
蛋白质印迹证实周细胞中TLR-4成功沉默，并持续7天。在MVEC/PC共培养物中观察到LPS对白蛋白清除率有剂量和时间依赖性效应。周细胞中TLR-4沉默的共培养消除了LPS剂量依赖性白蛋白清除率增加。
结论
TLR-4调节LPS暴露时周细胞介导的毛细血管渗漏。我们的TLR-4沉默模型可用于进一步研究TLR-4在周细胞介导的毛细血管渗漏中的作用。
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