Integrins inhibit LPS-induced DNA strand breakage in cultured lung endothelial cells.
Integrins inhibit LPS-induced DNA strand breakage in cultured lung endothelial cells.
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DOI：
10.1152/ajplung.1996.270.4.l689
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发表时间：
1996-04
期刊：
影响因子：
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通讯作者：
D. Hoyt;R. Mannix;M. Gerritsen;Simon C Watkins;J. Lazo;B. Pitt
中科院分区：
文献类型：
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作者：
D. Hoyt;R. Mannix;M. Gerritsen;Simon C Watkins;J. Lazo;B. Pitt
胶原抑制脂多糖（LPS）处理的绵羊肺动脉内皮细胞（SPAEC）的急性DNA链断裂和凋亡。在这里，我们测试了主要的基底膜成分，IV型胶原蛋白，层粘连蛋白和纤连蛋白，和整合素配体和抗整合素抗体，以抑制SPAEC和BALB/c小鼠肺内皮细胞（MLEC）的LPS引起的DNA断裂的能力。用末端脱氧核苷酸转移酶原位标记的DNA链断裂显示，在与1微克LPS/ml孵育后2小时内，附着的SPAEC和MLEC中的DNA断裂相似。在明胶、IV型胶原、层粘连蛋白、细胞纤连蛋白或血浆纤连蛋白上铺板的细胞中，急性DNA链断裂减少。将细胞接种在用整联蛋白配体六肽GRGDSP（40微克/cm 2）包被的表面上，而不是用GRADSP包被的表面上，也抑制了DNA断裂。LPS诱导的DNA链断裂在涂有鼠α 5-、β 1或β 3-整联蛋白亚基抗体的表面上的MLEC中受到抑制。在细胞单层上方的培养基中加入抗整联蛋白抗体（而非GRGDSP）可抑制链断裂。尽管有类似的急性DNA断裂，但MLEC比SPAEC表现出更少的脱离和细胞凋亡，这与这两种细胞类型中细胞凋亡的传感或处理系统的差异一致。这些结果表明，细胞外基质和整合素活化可以抑制LPS的遗传毒性。
Collagen inhibits acute DNA strand breakage and apoptosis in sheep pulmonary artery endothelial cells (SPAEC) treated with lipopolysaccharide (LPS). Here we tested the ability of major basement membrane components, type IV collagen, laminin and fibronectin, and integrin ligands and anti-integrin antibodies to inhibit DNA breakage caused by LPS in SPAEC and BALB/c murine lung endothelial cells (MLEC). In situ labeling of DNA strand breaks with terminal deoxynucleotidyl transferase revealed similar DNA breakage in attached SPAEC and MLEC within 2 h after incubation with 1 microgram LPS/ml. Acute DNA strand breakage was reduced in cells plated on gelatin, type IV collagen, laminin, cellular fibronectin, or plasma fibronectin. DNA breakage was also suppressed by plating cells on surfaces coated with the integrin ligand hexapeptide, GRGDSP (40 micrograms/cm2), but not with GRADSP. LPS-induced DNA strand breakage was inhibited in MLEC plated on surfaces coated with antibodies to murine alpha 5-, beta 1, or beta 3-integrin subunits. Addition of anti-integrin antibodies, but not GRGDSP, to the medium above cell monolayers inhibited strand breakage. Despite similar acute DNA breakage, MLEC exhibited less detachment and apoptosis than SPAEC, consistent with a difference in the sensing or processing systems for apoptosis in these two cell types. These results demonstrate that extracellular matrices and integrin activation can inhibit the genotoxicity of LPS.