软骨血管生成抑制因子抑制血管生成的研究

Inhibitory Effect fo Cartilage AngiogenesisInhibiting Factor on Angiogenesis.

小牛气管软骨经盐酸胍抽提，丙酮分级沉淀，膜超滤，柱层析等步骤得到软骨血管生成抑制因子（ｃａｒｔｉｌａｇｅａｎｇｉｏｇｅｎｅｓｉｓｉｎｈｉｂｉｔｉｎｇｆａｃｔｏｒ，ＣＡＩＦ）．ＳＤＳ－聚丙烯酰胺凝胶电泳显示ＣＡＩＦ由单一组分组成，分子量为２７７００．通过［￣３Ｈ］－ＴｄＲ掺入，活细胞检测等方法测定ＣＡＩＦ对内皮细胞、Ｈｅｌａ细胞、ＱＧＹ７７０３细胞与小鼠骨髓细胞、人皮肤成纤维细胞等的ＤＮＡ合成的影响，以及细胞毒作用．采用鸡胚绒毛尿囊膜实验测定ＣＡＩＦ对血管生成的抑制效应．结果显示：ＣＡＩＦ对内皮细胞产生强的抑制作用，对Ｈｅｌａ细胞抑制很弱，对ＱＧＹ７７０３细胞、小鼠骨髓细胞、人皮肤成纤维细胞均无抑制作用；对鸡胚绒毛尿囊膜的血管生成产生明显的抑制作用．提示ＣＡＩＦ能较特异地抑制血管生成，ＣＡＩＦ达到电泳纯，是专一性较强的血管生成抑制因子．

Cartilage angiogenesis inhibitingfactor(CAIF)was purified from cartilage byguanidine hydrochloride extraction,acetonefractional precipitation, ultrafiltration,col-umn chromatography.With SDS-PAGF,itshowed a single band of CAIF with molecularweight of 27 700.By the methods of [￣3H]-TdR incorporation and the determination ofmortality of cells,the effect of CAIF on theDNA synthesis and cytotoxicity of all celllines were investigated and the inhibitory ef-fect of CAIF on angiogenesis was measured bythe experiment of chorioallantoic membrane ofchicken embryos. The results showed thatCAIF inhibited the endothelial cell stronglyand Hela cell slightly; but not QGY7703 cell,human skin fibroblast, and mice myeloid cell.It also inhibited the angiogenesis in thechorioallantoic membrane of chicken embryossignificantly.All these results suggest thatCAIF has the inhibitory effect on angiogene-sis. It is a specific angiogenesis inhibiting fac-tor, and may be a useful drug on tumor thera-py in some day.