不同生长期壁虎水取物对肝癌细胞Bel-7402增殖抑制及诱导分化活性的影响  

Effects of Gecko Aqueous Extracts from Different Growth Stages on Proliferation Inhibition and Differentiation Induction Activities of Human Hepatoma Carcinoma Bel-7402 Cells

作　　者：王慧慧 孙艺茹 陈心怡 位雪 谢雪晴 王春梅 WANG Hui-hui;SUN Yi-ru;CHEN Xin-yi;WEI Xue;XIE Xue-qing;WANG Chun-mei(School of Life Science,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学生命科学学院,北京100029

出　　处：《特产研究》2021年第3期1-5,18,共6页Special Wild Economic Animal and Plant Research

基　　金：国家自然基金青年科学基金项目(81774014);北京中医药大学新奥奖励基金课题(2016XAJLJJ-015);北京中医药大学发展基金(2020071420021);中央高校基本科研业务费专项资金资助(2020-JYB-ZDGG-054)。

摘　　要：本文研究不同生长时期鲜无蹼壁虎水提粗蛋白的抗肿瘤活性差异,为确定其最适入药期提供理论依据。选取Ⅰ(3月龄)、Ⅱ(5月龄)、Ⅲ(8月龄)、Ⅳ(1年龄)和Ⅴ(3年龄)共5种生长期的壁虎,观测体型差异,制得鲜壁虎水提物,通过BCA法测量样品的总蛋白质浓度,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)法检测可溶性蛋白质分子量分布多样性;通过MTT法评估用药24、48、72 h后5种体型壁虎对人肝癌Bel-7402细胞的细胞增殖活性的影响;通过磷酸苯二钠微板法检测癌细胞碱性磷酸酶(ALP)活力,反映其诱导分化活性。结果显示,5组壁虎样品单只体长和体重分布为6.1~12.5 cm/0.59~6.3 g,随着体型及年龄增大,水提物可溶性蛋白含量占比有所减少,蛋白种类依分子量不同表现出20~25 k Da、30 kDa、40~65 kDa,区间显著差异;肿瘤细胞增殖抑制活性与壁虎年龄、用药时间和浓度呈正相关,、Ⅳ、Ⅴ相较Ⅰ、Ⅱ表现出更好的抑制活性,分别为86.45%(Ⅲ)、88.94%(Ⅳ)和82.94%(Ⅴ),较强的Bel-7402细胞ALP活力抑制性,分别为52.62%(Ⅲ)、56.98%(Ⅳ)和56.90%(Ⅴ)。结果表明,壁虎成长至8月龄时,抗肿瘤活性达到最高且伴随进一步成熟再无明显差异,视为最佳入药采收期。To study the anti-tumor effect of AG(aqueous extracts of fresh gecko)at different growth stages and to provide a theoretical basis for confirming an optimal recovery period.Five kinds of samples:Ⅰ(three-month-old),Ⅱ(five-month-old),(eight-month-old),Ⅳ(oneyear-old),Ⅴ(three-year-old)for this study were collected.The difference in body shape,weight of fresh gecko were measured.The total protein concentration of AG was detected by BCA assay.The cytotoxic activity of AG on human liver cancer Bel-7402 cells was evaluated by MTT assays.The soluble protein diversity was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).The cytotoxic activity of AG on Bel-7402 cells was evaluated by MTT assays with the same crude drug content for 24 h,48 h,72 h.ALP(alkaline phosphatase)was detected by biochemical methods with commercial kits.The results showed that the body length and body weight distribution of the sample were 6.1~12.5 cm/0.59~6.3 g.The content of soluble proteins in the five aqueous extracts decreased with the increase of body size,and the protein types showed significant differences depending on the molecular weight.Concentrated in the range of20~25 k D,30 kD and 40~65 kD;The anti-tumor activity increased gradually,and it was correlated positively with the time and concentration of the drug,,Ⅳ,ⅤCompared withⅠandⅡshowed better inhibitory activity,and the highest inhibition rates of them were 86.45%(Ⅲ),88.94%(Ⅳ),and 82.94%(V),respectively,and showed strong inhibition of ALP activity in Bel-7402 cells,which were 52.62%(Ⅲ),56.98%(Ⅳ),56.90%(V).It was concluded that when the gecko grows to 8 months old,the anti-tumor activity was the highest and there was no significant difference in further maturity,so it was regarded as the best harvesting period.

关 键 词：壁虎 生长期 水提物 抗肿瘤活性 

分 类 号：R285[医药卫生—中药学] R282.74[医药卫生—中医学]