[关键词]
[摘要]
目的研究魟鱼软骨多糖(raycartila geglycosam inoglycans,RCG)的提取、分离与纯化方法,并初步探讨其生物活性。方法采用盐酸胍提取、丙酮分级沉淀、膜超滤、Sephadex凝胶柱色谱分离、纯化获得魟鱼软骨多糖,由HPLC确定其相对分子质量和质量分数;建立Lewis肺癌小鼠模型,将实验分为生理盐水(Saline)组、不同剂量(500、250、125mg/kg)鱼软骨多糖组、环磷酰胺(CTX,60mg/kg)组,观察小鼠肿瘤生长情况,绘制肿瘤生长曲线,计算原发瘤与肺转移灶数抑瘤率,免疫组织化学染色计数肿瘤组织微血管密度(MVD),采用RT-PCR检测瘤组织血管内皮生长因子(VEGF)基因表达。结果经HPLC检测,该组分为单一多糖,其质量分数达99%以上。生物活性检测结果表明,与生理盐水组比较,该多糖各剂量组小鼠的肿瘤生长曲线平缓,原发瘤抑瘤率、肺转移灶数与生理盐水组比较差异显著(P<0.05、0.01);与生理盐水组比较,鱼软骨多糖各剂量组MVD显著减少、VEGFmRNA表达水平显著降低(P<0.01)。结论鱼软骨多糖明显抑制小鼠Lewis肺癌原发瘤的生长和转移,并可抑制肿瘤的血管形成。
[Key word]
[Abstract]
Objective To explore the methods of extraction, isolation, purification, and biological activities of ray cartilage glycosaminoglycans (RCG). Methods RCG was purified by guanidine hydrochlorid extraction, acetone fractional precipitation, ultrafiltration, and Sephadex column chromatography. The purity and molecular mass of RCG were measured by means of HPLC. The model of mouse with Lewis lung carcinoma was made, the experimental mice were randomly divided into normal saline group, RCG (500, 250, and 125 mg/kg) groups, and CTX (60 mg/kg) group. Tumor growth states of mice were observed, tumor growth curve was described, inhibitory rates of primary tumor and number of lung metastasis focus were measured; microvessel density (MVD) was quantitated by immunohistochemistry using monoclonal antibodies of CD31; the expression of vascular endothelial growth factor (VEGF) mRNA was determined with RT-PCR. Results Using HPLC, a single glycosaminoglycans with molecular mass 9.7×104 was collected and its purity exceeded ninty-nine percent. Tumor growth curves in RCG groups were smooth compared with saline group. There were significant differences of inhibitory rates of primary tumor, number of lung metastasis focus and MVD between RCG groups and saline group. VEGF mRNA expression levels in RCG groups were reduced significantly compared with saline group. Conclusion RCG could effectively inhibit the growth and metastasis of primary Lewis lung carcinoma in C57BL/6 mouse and angiogenesis.
[中图分类号]
[基金项目]
辽宁省教育厅基金资助项目(202173371)