3种壁虎酶解物对H22荷瘤小鼠及Bel7402肝癌细胞抑制活性差异的研究  

Anti-tumor effect of 3 gecko hydrolysates on H22 transplanted tumor in mice and Bel7402 cells

作　　者：孙艺茹 王春梅 谢雪晴 王慧慧 SUN Yi-ru;WANG Chun-mei;XIE Xue-qing;WANG Hui-hui(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 100029)

机构地区：[1]北京中医药大学生命科学学院,北京100029

出　　处：《中南药学》2020年第7期1132-1136,共5页Central South Pharmacy

基　　金：北京中医药大学新奥奖励基金课题(No.2019);中央高校基本科研业务费专项资金(No.2020-JYB-ZDGG-054);国家自然科学基金项目(No.81774014)。

摘　　要：目的研究3种解壁虎酶解物对其体内外抗肿瘤活性的影响。方法采用木瓜蛋白酶(papain)、碱性蛋白酶2.4L(alcalase)、胰蛋白酶(trypsin)分别水解壁虎粉,得水溶性粗肽混合物。采用甲醛滴定法测定游离氨基态氮(AN)含量,从而计算水解度(DH);MTT法评估水解产物对人肝癌Bel7402细胞体外增殖活性的影响;制备H22荷瘤小鼠模型,检测壁虎不同酶解终产物的抑瘤效果及其对荷瘤小鼠体重和免疫器官的影响。结果Alcalase水解效率较高,水解度可达63%;体外抗肿瘤活性排序为alcalase(79.91%)>papain(67%)>原粉组(49.37%);体内抗肿瘤活性为alcalase(67.11%)>papain(53.48%)>原粉组(17.47%),trypsin组体内外活性与原粉组相比均无显著差异。结论壁虎酶解物体内外抗肿瘤活性具有一致性且与水解度相关,alcalase酶解效果最优。Objective To determine the anti-tumor effects of 3 gecko hydrolysates in vitro and in vivo.Methods Gecko powder was hydrolyzed by 3 pure enzymes(papain,alcalase 2.4L,and trypsin)respectively to obtain the water-soluble crude peptide mixture.The formaldehyde titration method was used to determine the content of free amino nitrogen to calculate the degree of hydrolysis.MTT method was applied to evaluate the proliferation of hydrolysates to human liver cancer Bel7402 cells in vitro.The H22 tumor-bearing mouse models were established.The tumor suppression of different products and changes in immune organ indexes of the mice were detected.Results Alcalase showed the highest enzymatic hydrolysis efficiency up to 63%;the anti-tumor activity in vitro was alcalase(79.91%)>papain(67%)>original gecko powder(49.37%),and the anti-tumor activity in vivo was alcalase(67.11%)>papain(53.48%)>original gecko powder(17.47%).There was no significant difference between the activities of trypsin group and original gecko powder group in vitro or in vivo.Conclusion The anti-tumor activity of gecko hydrolysates in vivo and in vitro are consistent,and is related to the degree of hydrolysis with the best enzymatic effect alcalase.

关 键 词：壁虎 酶解 碱性蛋白酶 木瓜蛋白酶 胰蛋白酶 H22荷瘤小鼠 抗肿瘤活性 

分 类 号：R932[医药卫生—生药学] R286[医药卫生—药学]