烟台刺参酶解工艺最佳用酶初筛及体外抗肿瘤研究
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摘要
通过考察不同酶对海参酶解产物的体外抗肿瘤活性及水解度的影响,确定最佳用酶,为海参抗肿瘤研究提供新思路。以海参水提液及100μg/mL顺铂为对照组,对比不同提取条件下海参水提液对喉癌Hep-2细胞的增殖抑制率,结合茚三酮显色法测定海参酶解液水解度,确定最佳用酶。结果显示:弹性蛋白酶、胰蛋白酶及胰凝乳蛋白酶处理组水解度分别为10.05%、7.86%、1.54%,其IC_(50)分别为0.642、0.763、0.917 mg/mL,胃蛋白酶处理组水解度为2.72%,抗肿瘤效果极差。说明海参酶解最佳用酶为弹性蛋白酶,其次是胰蛋白酶及胰凝乳蛋白酶。
By investigating the effects of different enzymes on the antitumor activity and hydrolysis degree of enzymatic hydrolysates of sea cucumber in vitro,the optimum enzymes were screened out to provide new ideas for the study of sea cucumber antitumor.Taking sea cucumber water extract and cisplatin as control group,the inhibition rates of sea cucumber water extracts on the proliferation of laryngeal cancer Hep-2 cells under different extraction conditions were compared.The hydrolysis degree of sea cucumber enzymatic hydrolysate was determined by Ninhydrin chromogenic method,and the optimum enzyme was determined.The results showed that the hydrolysis degrees of elastase,trypsin and chymotrypsin were 10.05%,7.86%and 1.54%,respectively,and the IC50 of them were 0.642,0.763,0.917 mg/mL.The hydrolysis degree of pepsin treatment was 2.72%,and the anti-tumor effect was very poor.The results showed that elastase was the best enzyme for enzymatic hydrolysis of sea cucumber,followed by trypsin and chymotrypsin.
By investigating the effects of different enzymes on the antitumor activity and hydrolysis degree of enzymatic hydrolysates of sea cucumber in vitro,the optimum enzymes were screened out to provide new ideas for the study of sea cucumber antitumor.Taking sea cucumber water extract and cisplatin as control group,the inhibition rates of sea cucumber water extracts on the proliferation of laryngeal cancer Hep-2 cells under different extraction conditions were compared.The hydrolysis degree of sea cucumber enzymatic hydrolysate was determined by Ninhydrin chromogenic method,and the optimum enzyme was determined.The results showed that the hydrolysis degrees of elastase,trypsin and chymotrypsin were 10.05%,7.86%and 1.54%,respectively,and the IC50 of them were 0.642,0.763,0.917 mg/mL.The hydrolysis degree of pepsin treatment was 2.72%,and the anti-tumor effect was very poor.The results showed that elastase was the best enzyme for enzymatic hydrolysis of sea cucumber,followed by trypsin and chymotrypsin.
引文
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[3]何丽霞,李勇.海参肽生物学功能研究进展[J].食品科学,2015,36(09):215-218.
[4]赵玲,殷邦忠,刘淇,等.4种海参多肽抗氧化活性的比较研究[J].中国海洋药物,2012,31(02):19-24.
[5]张伟伟,陆茵.海参的抗肿瘤作用研究进展[J].中华中医药杂志,2010,25(01):105-108.
[6]Bulysheba V S,Maslova M G,Boikova EA,et al.Effect of the Japanese cucumaria on phospholipid metabolism in the liver of mice with solid Ehrlich carcinoma[J].Eksp Onkol,1986,8:70(from Medline).
[7]王肇扬,王彦多,曹广超,苑文姝,成丽媛,张珊珊,王集会.鼠妇粉酶解最佳用酶的初步筛选[J].当代化工,2018,47(06):1151-1154.
[8]吕迎兰,刘颖,王彦多,毛会秀,曹广超,王集会,高世杰.地龙粉酶解用酶初步筛选[J].山东化工,2018,47(05):20-22+25.
[9]刘金虎,王哲,曹广超,等.蛴螬粉酶解工艺最佳用酶的初步筛选及体外抗肿瘤活性的研究[J].华西药学杂志,2018,33(04):454-456.
[10]刘颖,朱晓明,王彦多,曹广超,施东方,吕迎兰,王集会,史磊.正交实验优化美洲大蠊粉混合酶酶解工艺[J].当代化工,2018,47(06):1131-1134.
[11]刘春雨,张盼盼,王颖,等.蜈蚣粉酶解工艺最佳用酶的初步筛选[J].中南药学,2017,15(06):759-762.
[12]王彦多,方园,刘颖,等.蜈蚣酶解工艺优化及抗肿瘤活性研究[J].四川中医,2018,36(10):49-52.
[13]曹广超,王彦多,刘颖,李亚男,刘金虎,桑晓,毛会秀,王集会,史磊.发酵蜣螂粉乙醇提取物抗肿瘤活性研究[J].山东化工,2018,47(02):22-24.
[14]桑晓,张瑞,王彦多,毛会秀,史磊.野生与养殖全蝎酶解后主要成分含量及抗肿瘤作用的对比研究[J].中南药学,2018,16(02):171-174.
[15]王悦,容蓉,巩丽丽.解表类中药抗病毒作用研究进展[J].中南药学,2018,16(02):192-194.
[16]岳静.薏苡仁及其组分对类风湿关节炎大鼠抗炎作用研究[D].山东中医药大学,2017.
[17]王振立,刘桂敏,郑瑞等.刺参酸性粘多糖抑制小鼠肿瘤细胞DNA合成及其代谢研究[J].中国医药工业杂志,1993(09):405-408.