摘要
以赤缸软骨为原料，对赤缸软骨中新生血管抑制组分的提取方法及其生物学活性进行了初步研究。采用盐酸胍抽提、冷冻离心、透析、冷冻干燥等方法，得到分子量为3～300kDa的活性组分，活性鉴定采用改进的Robert法观察赤缸软骨提取物对鸡胚绒毛尿囊膜（CAM）血管生成的作用情况。赤缸软骨提取物活性给药组为质量分数为109／6、19／6、0．1％的甲基纤维素溶液，空白对照组为质量分数为10％的MC溶液，加样24h后对各给药组样品给药部位进行形态学观察，并通过计算机图像分析软件原理编程进行血管数量统计。试验结果表明：赤缸软骨提取物具有抑制新生血管形成的作用，其活性与给药组质量分数成正比。且软骨血管生成抑制因子直接来源于动物组织，无明显毒副作用，在肿瘤治疗方面可能具有良好的应用前景。
Anti-angiogenesis therapy to inhibit tumor growth has aroused the interest of many researchers. Shark cartilage angiogenesis inhibitor factor （SCAIF） extracted from shark＇s cartilage has strong angiogenesis inhibitory activity. Pharmacopeia records that Dasyatis akajei has the same effect as shark, but it has much higher resources than shark. Extracting method of anti-angiogenesis component from Dasyatis akajei cartilage and its bioactivity is discussed. Using 1. 0 mol/L guanidine hydrochloride and 0. 02 mol/L MES solution （containing 0. 02% EDTA）, the material was vibrated and extracted for 48 h at 28℃ and then centrifugalized for 30 min at 4 ℃. The supernatant was centrifugalized again at the same condition, and then pour the interception supernatant into a dialysis bag with molecular weight of 3 kDa to be dialysed for 24 h in 0.02 mol/L Tris-HCl buffer of pH 7.6, and dialysed again in a dialysis bag with molecular weight 300 kDa for another 24 h to obtain the interception with molecular weight 3-300 kDa. Freeze drying of the interception resulted in the DCGE （Dasyatis akajei cartilage guanidine hydrochloride extract） of molecular weight 3-300 kDa from the Dasyatis akajei cartilage. Improved Robert method was used to identify the activity of DCGE on CAM （chicken embryo chorioallantoic membrane）. DCGE treatment groups were given 10%, 1%,0.1% of the methyl cellulose solution and the control group was given 10% of the MC solution. After 24 hours morphological observation was made on all the groups and the number of blood vessels was investigated with computer image analysis system（CIAS）. The results show that CAM membrane of the control group was fused with carrier and some large blood vessels ran through it and grew well. Light-coloured spots resulted from less blood vessels appeared on all the groups given DCGE of low, middle and high dosage. Blood vessels in the spots reduced in accordance with the cartilage extract from low to high concentration, which shows that DCAI from Dasyatis akajei cartilage has evident angiogenesis inhibiting effect, and there is a positive relation between the concentration and inhibiting effect. The statistics of new angiogenesis number in the point sample and the significant difference analysis indicates that no matter the area of blood vessels or the ratio of vessels areas, three groups given different concentrations of DCGE all have a significant difference from the control group in accordance with the descending order of concentration, the ratio of vessels areas （white/ Black） has an increasing trend. The result of the present study indicates that DCGE has an angiogenesis inhibitory effect, which is in direct proportion with the concentration. DCAI which is directly from animal tissues and has no significant side effects, have a good prospect in tumor therapy.
出处
《海洋学研究》
北大核心
2008年第4期61-65,共5页
Journal of Marine Sciences
基金
浙江省科技厅2004年科研资助项目(2004C33097)
浙江省新苗人才计划资助项目(2207G60G2110004)
关键词
赤缸软骨
血管抑制组分
鸡胚绒毛尿囊膜
Dasyatis akajei cartilage
angiogenesis inhibitory components
chicken chorioallantoic membrane
