文档介绍：新疆医科大学
硕士学位论文
鹰嘴豆芽提取物诱导Caco-2细胞凋亡作用机制的研究
姓名:毛人杰
申请学位级别:硕士
专业:免疫学
指导教师:丁剑冰
2011-04
新疆医科大学硕士学位论文
鹰嘴豆芽提取物诱导 Caco-2 细胞凋亡作用机制的研究
研究生:毛人杰导师:丁剑冰教授
摘要
目的:本实验研究鹰嘴豆芽乙醇提取物(CSE)对人结肠腺癌 Caco-2 细胞增殖
及细胞凋亡的影响和机制,并为鹰嘴豆应用于结肠癌的临床治疗和预防提供实验依
据。方法:鹰嘴豆芽的活性成分提取和含量测定;MTT 法观察鹰嘴豆芽提取物对
Caco-2 细胞增殖的影响;透射电镜观察细胞超微结构;琼脂糖凝胶电泳法进行细胞
凋亡的 DNA 分析;流式细胞术检测凋亡情况;蛋白印迹法检测鹰嘴豆芽提取物干预
后 Caco-2 细胞的 Bcl-2 和 Bax的表达量;化学发光法检测 Caco-2 细胞的 Caspase3/7
酶活性。结果:CSE 含 ± %皂苷,± %异黄酮;不同浓度的 CSE
对 Caco-2 细胞均有一定的增殖抑制作用,且呈量效和时效关系;电镜观察到细胞出
现染色质边移和凋亡小体;DNA 电泳呈现梯度条带,表明 CSE 可诱导 Caco-2 细胞
凋亡;3、5、10 µg/mL 的 CSE作用后,细胞凋亡率分别为 ± % (P<)、
± % (P<)、±% (P<)。鹰嘴豆芽提取物能降低 Caco-2 细胞
Bcl-2 蛋白的表达量,提高 Bax 蛋白的表达量,使 Bcl-2/Bax 比值下降,并增强
Caspase3/7 酶活性(P<)。结论:CSE 可通过诱导细胞凋亡而抑制人结肠腺癌株
Caco-2 细胞的生长,其机制可能与下调 Bcl-2 蛋白,上调 Bax蛋白,提高 Caspase3/7
酶活性有关。
关键词:鹰嘴豆;Caco-2 细胞;细胞凋亡;Bcl-2;Bax

1
新疆医科大学硕士学位论文
The mechanism underlying apoptosis-inducing effects of
chickpea sprout extract on Caco-2 cells
Postgraduate: Mao Renjie Supervisor: prof. Ding Jianbing
Abstract
Objective: To explore the effect of chickpea sprout ethanol extract (CSE) on proliferation
and apoptosis of human colon adenocarcinoma cell line Caco-2 in vitro and underlying
mechanism. It provides evidence for the application of chickpea in clinical treatment of
human colon adenocarcinoma. Methods: CSE was produced and ponent was
determined. The proliferation inhibition rate of Caco-2 cells was measured by MTT assay.
The ultrastructure of Caco-2 cells was observed under the electron microscope. Apoptosis
was assayed by agarose gel eletrophoresis, and cell apoptosis rate was evaluated by flow
cytometry. The protein expression of Bcl-2 and Bax were examined by Western blotting;
The activities of caspase-3 and caspase-7 was determined by Caspase-3/7 Luminescence
Assay. Results: CSE contained ±% of saponins and ±% of isoflavones.
CSE inhibited the Caco-2 cell p