INTERSTITIAL COLLAGENASE IS REQUIRED FOR ANGIOGENESIS IN-VITRO
INTERSTITIAL COLLAGENASE IS REQUIRED FOR ANGIOGENESIS IN-VITRO
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DOI：
10.1006/dbio.1994.1104
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发表时间：
1994-04-01
影响因子：
2.7
通讯作者：
MITCHELL, MA
中科院分区：
文献类型：
--
作者：
FISHER, C;GILBERTSONBEADLING, S;MITCHELL, MA
人脐静脉内皮细胞（HUVEC）侵入胶原凝胶，并在佛波醇酯刺激后在凝胶内建立血管样结构。通过测量由[H-3]胶原组成的凝胶的放射性释放来定量该过程。胶原蛋白稳定降解了几个星期的佛波酯处理的细胞，而没有收到佛波酯的细胞胶原蛋白溶解很少注意到。基质金属蛋白酶（MMPs）分泌的HUVECs的检查显示了显着的诱导间质胶原酶。成熟形式的明胶酶A的产生也受到刺激，明胶酶B的分泌也受到刺激。未检测到溶基质素。两种MMPs抑制剂，天然存在的金属蛋白酶组织抑制剂（TIMP; 10 μ M/ml）和合成的肽抑制剂BB-94（1 μ M）都能有效阻断HUVEC介导的胶原降解。对照、PMA处理的HUVEC以及接受TIMP或BB-94的PMA处理的HUVEC的形态学检查显示，MMP抑制导致胶原凝胶内的侵袭和小管形成的阻断。MMP表达和抑制小管形成在体外与人真皮微血管内皮细胞获得了类似的结果。胶原蛋白水解片段的检查显示BB-94和TIMP均阻断I型胶原α 1和α 2链的裂解以及原胶原片段A和B的出现，表明抑制剂直接作用于间质胶原酶。我们的研究结果表明，间质胶原酶是体外血管生成所必需的。(C)1994年出版社出版。
Human umbilical vein endothelial cells (HUVECs) invade collagen gels and establish vascular-like structures within the gel following stimulation with phorbol esters. This process was quantitated by measuring release of radioactivity from gels composed of [H-3]collagen. Collagen was steadily degraded over the period of several weeks by phorbol ester-treated cells while little collagenolysis by cells not receiving phorbol ester was noted. Examination of matrix metalloproteinases (MMPs) secreted by HUVECs revealed a prominent induction of interstitial collagenase. Production of the mature forms of gelatinase A was also stimulated, as was the secretion of gelatinase B. Stromelysin was not detected. Two inhibitors of MMPs, the naturally occurring tissue inhibitor of metalloproteinases (TIMP; 10 mug/ml) and the synthetic, peptide inhibitor BB-94 (1 muM) were both effective at blocking HUVEC-mediated collagen degradation. Morphological examination of control, PMA-treated HUVECs, as well as PMA-treated HUVECs receiving TIMP or BB-94, revealed that MMP inhibition resulted in a block to invasion and tubule formation within the collagen gels. Similar results for MMP expression and inhibition of tubule formation in vitro were obtained with human dermal microvascular endothelial cells. Examination of collagen proteolytic fragments revealed that both BB-94 and TIMP blocked cleavage of the alpha1 and alpha2 chains of type I collagen and the appearance of tropocollagen fragments A and B, demonstrating that the inhibitors were acting directly upon interstitial collagenase. Our results demonstrate that interstitial collagenase is required for angiogenesis in vitro. (C) 1994 Academic Press, Inc.