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目的:对微波辅助碱液提取孔鳐软骨多糖的方法及其抗氧化和血管生成抑制活性进行研究。方法:采用微波辅助碱液提取与木瓜蛋白酶水解相结合的加工工艺,以正交试验法优化提取工艺参数。通过还原力、DPPH·和·OH清除实验对多糖抗氧化能力进行测定。采用鸡胚绒毛尿囊膜(CAM)模型对其血管生成抑制活性进行评价。结果:微波辅助碱液提取最佳工艺参数是:微波功率650W、提取温度60℃、碱质量分数15%、液料比35:1、提取时间7min。木瓜蛋白酶脱蛋白精制孔鳐软骨多糖的最佳条件是:酶解温度50℃,pH7.0,酶解时间2h,酶量1.0%。在此条件下精制的多糖为白色粉末,得率13.77%,纯度88.75%。在相同浓度下精制孔鳐软骨多糖抗氧化能力均大于粗提多糖。精制多糖的血管生成抑制活性与硫酸软骨素(CS)相当,并且呈剂量依赖关系。结论:微波辅助碱液提取和木瓜蛋白酶脱蛋白所得精制多糖具有很好的抗氧化和血管生成抑制活性。
OBJECTIVE: To study the method for extracting polysaccharides of Pterygium chondrocytes by microwave-assisted alkaline extraction and its anti-oxidative and anti-angiogenic activity. Methods: The microwave assisted extraction of lye with papain hydrolysis process combined with orthogonal test method to optimize the extraction process parameters. Antioxidant capacity of polysaccharides was determined by reducing force, DPPH · and · OH scavenging experiments. The chick embryo chorioallantoic membrane (CAM) model was used to evaluate its angiogenesis inhibitory activity. Results: The optimal extraction parameters of microwave-assisted alkaline extraction were: microwave power 650W, extraction temperature 60 ℃, alkali mass fraction 15%, liquid to material ratio 35: 1 and extraction time 7min. The optimal conditions of papain deproteinization were as follows: enzymolysis temperature 50 ℃, pH7.0, hydrolysis time 2h, enzyme amount 1.0%. Refined polysaccharide in this condition is a white powder with a yield of 13.77% and a purity of 88.75%. In the same concentration, the anti-oxidation ability of the purified pericarpium cartilage polysaccharide is greater than the crude polysaccharides. The inhibitory activity of purified polysaccharides on angiogenesis was comparable to that of chondroitin sulfate (CS) in a dose-dependent manner. CONCLUSION: The polysaccharide extracted by microwave-assisted alkaline extraction and papain deproteinization has good antioxidant and angiogenic inhibitory activity.