Toll-like receptor-4 message is up-regulated in lipopolysaccha ride-exposed rat lung pericytes
Toll-like receptor-4 message is up-regulated in lipopolysaccha ride-exposed rat lung pericytes
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DOI：
10.1016/j.jss.2006.03.007
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发表时间：
2006-07-01
影响因子：
2.2
通讯作者：
Steffes, Christopher
中科院分区：
文献类型：
--
作者：
Edelman, David A.;Jiang, Yang;Steffes, Christopher
背景周细胞是多功能的、多态的血管周细胞，位于微血管基底层内，位于内皮细胞的近腔侧，并且被认为在脓毒症中观察到的毛细血管渗漏中起调节作用。Toll样受体4（TLR-4）在脂多糖（LPS）诱导的脓毒症的活化过程中被认为是LPS/CD 14复合物的近端跨膜受体。我们假设TLR-4存在于肺周细胞上并且响应于LPS而上调。分离培养大鼠肺微血管周细胞。使用来自第3-5代的细胞并用LPS（对照、10 ng/mL和100 ng/mL）处理18小时。进行免疫染色和免疫印迹以检测CD-14、TLR-2和TLR-4的存在。实时荧光定量PCR检测CD-14、TLR-2、TLR-4 mRNA的表达。免疫染色和免疫印迹显示每个处理组的周细胞中存在CD-14、TLR-2和TLR-4，实时聚合酶链反应证实存在CD-14、TLR-2和TLR-4的mRNA。在治疗后4 h，在存在增加的LPS的情况下，观察到CD-14、TLR-2和TLR-4的mRNA增加。在LPS处理后18 h，注意到mRNA的减少。TLR-4在LPS增加的情况下的上调表明其在周细胞LPS诱导的活化中的重要性。外周细胞TLR-4对LPS的识别可能在脓毒症中观察到的毛细血管渗漏中起作用。这些数据还表明，周细胞，一旦被认为是被动参与炎症级联反应，可能是积极的成员。(c)2006爱思唯尔公司All rights reserved.
Background. Pericytes are multifunctional, polymorphic perivascular cells that lie within the microvessel basal lamina, are located on the abluminal side of endothelial cells, and are thought to play a regulatory role in capillary leak observed in sepsis. Toll-Like receptor 4 (TLR-4) has been implicated as the proximal transmembrane receptor for the LPS/CD 14 complex during the activation of lipopolysacharide (LPS)-induced sepsis. It is our hypothesis that TLR-4 is present on lung pericytes and is up-regulated in response to LPS.Methods. Rat microvascular lung pericytes were isolated and cultured. Cells from passage 3-5 were used and treated with LPS (control, 10 ng/mL, and 100 ng/mL) for 18 h. Immunostaining and immunoblotting were performed to detect the presence of CD-14, TLR-2, and TLR-4. Real-time polymerase chain reaction was used to analyze the presence and quantity of mRNA for CD-14, TLR-2, and TLR-4.Results. Immunostaining and immunoblotting revealed the presence of CD-14, TLR-2, and TLR-4 in pericytes from each treatment group, and real-time polymerase chain reaction confirmed the presence of mRNA for CD-14, TLR-2, and TLR-4. An increase in the mRNA was observed in CD-14, TLR-2, and TLR-4 in the presence of increasing LPS 4 h after treatment. At 18 h after LPS treatment, a decrease in mRNA was noted.Conclusions. The up-regulation of TLR-4 in the presence of increasing LPS suggests its importance in pericyte LPS-induced activation. Pericyte TLR-4 recognition of LPS could play a role in capillary leak seen in sepsis. These data also demonstrates that pericytes, once thought to be passive participants in the inflammatory cascade, may be active members. (c) 2006 Elsevier Inc. All rights reserved.