Horton M A, Dorey E L, Nesbitt S A, Samanen J, Ali F E, Stadel J M, Nichols A, Greig R, Helfrich M H
Department of Haematology, St. Bartholomew's Hospital, London, England.
J Bone Miner Res. 1993 Feb;8(2):239-47. doi: 10.1002/jbmr.5650080215.
This study details the investigation of induction of retractile shape change in the osteoclast through inhibition of adhesion between osteoclasts and matrix with (1) peptide analogs bearing an Arg-Gly-Asp (RGD) sequence, (2) antibodies to the integrin alpha V beta 3 vitronectin receptor, and (3) the RGD-containing snake venom peptide echistatin. Osteoclast retraction on dentin has been demonstrated for GRGDSP peptide, in contrast to the inactivity of the analog containing the conservative RGE sequence modification. An osteoclast adhesion assay employing rat or chick bone cells and serum-coated glass coverslips as substrate was developed for routine evaluation of inhibition of adhesion. Antibodies F4 and F11 to the beta 3 chain of rat vitronectin receptor were effective at submicromolar concentrations in rat osteoclasts (IC50 0.29 and 0.05 microM, respectively), whereas MAb 23C6 to human/chick vitronectin receptor was somewhat less effective against chick osteoclasts (IC50 1.6 microM). A rank order of RGD analog activity (mean IC50, microM) in the serum-coated glass adhesion assay was derived for the linear peptides GRGDSP (201 microM), GRGDTP (180 microM), Ac-RGDS-NH2 (84 microM), Ac-RGDV-NH2 (68 microM), RGDV (43 microM), GRGDS (38 microM), and RGDS (26 microM). The two most potent short peptides were the cyclic analog SK&F 106760 Ac-S,S-cyclo-(Cys-(N alpha Me)Arg-Gly-Asp-Pen)-NH2 (IC50 7.0 microM), and the Telios peptide H-Gly-S,S-cyclo-(Pen-Gly-Arg-Gly-Asp-Ser-Pro-Cys)-Ala-OH (IC50 6.6 microM). The snake venom peptide echistatin was the most potent substance evaluated in the serum-coated glass assay (IC50 0.78 nM) employing either rat or chick osteoclasts.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究详细阐述了通过以下方式抑制破骨细胞与基质之间的黏附，从而诱导破骨细胞发生可收缩形状变化的研究：（1）带有精氨酸 - 甘氨酸 - 天冬氨酸（RGD）序列的肽类似物；（2）整合素αVβ3玻连蛋白受体的抗体；（3）含RGD的蛇毒肽echistatin。与含保守RGE序列修饰的类似物无活性形成对比的是，已证实GRGDSP肽可使破骨细胞在牙本质上发生回缩。开发了一种以大鼠或鸡骨细胞以及血清包被的玻璃盖玻片为底物的破骨细胞黏附试验，用于常规评估黏附抑制情况。针对大鼠玻连蛋白受体β3链的抗体F4和F11在大鼠破骨细胞中，亚微摩尔浓度时就有效（IC50分别为0.29和0.05微摩尔），而针对人/鸡玻连蛋白受体的单克隆抗体23C6对鸡破骨细胞的效果稍差（IC50为1.6微摩尔）。在血清包被玻璃黏附试验中得出了RGD类似物活性的排序（平均IC50，微摩尔），线性肽GRGDSP（201微摩尔）、GRGDTP（180微摩尔）、Ac - RGDS - NH2（84微摩尔）、Ac - RGDV - NH2（68微摩尔）、RGDV（43微摩尔）、GRGDS（38微摩尔）和RGDS（26微摩尔）。两种最有效的短肽是环类似物SK&F 106760 Ac - S,S - 环 -（半胱氨酸 -（Nα甲基）精氨酸 - 甘氨酸 - 天冬氨酸 - 青霉胺）- NH2（IC50为7.0微摩尔）和Telios肽H - 甘氨酸 - S,S - 环 -（青霉胺 - 甘氨酸 - 精氨酸 - 甘氨酸 - 天冬氨酸 - 丝氨酸 - 脯氨酸 - 半胱氨酸）- 丙氨酸 - OH（IC50为6.6微摩尔）。在使用大鼠或鸡破骨细胞的血清包被玻璃试验中，蛇毒肽echistatin是评估的最有效物质（IC50为0.78纳摩尔）。（摘要截断于250字）
