药桑椹多糖组成分析及诱导肿瘤细胞凋亡研究  

COMPOSITION ANALYSIS OF POLYSACCHARIDES FROM FRUITS OF BLACK MULBERRY AND THE APOPTOSIS INDUCING ACTION IN TUMOR CELLS

作　　者：余婉莎 陈虎[1] 陈果[1] 向仲怀[1] 何宁佳[1] YU Wan-sha CHEN Hu CHEN Guo XIANG Zhong-huai HE Ning-jia(State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China)

机构地区：[1]家蚕基因组生物学国家重点实验室西南大学,重庆400715

出　　处：《营养学报》2017年第3期287-293,共7页Acta Nutrimenta Sinica

基　　金：重庆市科委院士基金;重庆市基础与前沿研究计划院士专项(No.cstc2016jcyjys0002)

摘　　要：目的分离鉴定药桑椹多糖(polysaccharide of black mulberry fruits,PBMF)及其单糖组成,并考察其抗氧化活性以及对He La细胞增殖和凋亡的影响,为药桑椹多糖的抗肿瘤研究及开发利用提供一定的理论依据。方法利用超高效液相色谱法(UPLC)分析药桑椹多糖的单糖组分;体外抗氧化实验观察PBMF的还原力、清除羟自由基能力;MTT法检测细胞增殖活性;倒置显微镜观察细胞形态变化;Annexin V-FITC/PI双染荧光显微镜观察He La细胞凋亡和坏死情况;流式细胞仪检测细胞周期和细胞凋亡;Western blot检测非活化态半胱氨酸天冬氨酸蛋白酶-3(procaspase-3)及半胱氨酸天冬氨酸蛋白酶-3(caspase-3)表达量。结果 PBMF单糖组成主要为甘露糖、鼠李糖、葡萄糖醛酸、葡萄糖、木糖、半乳糖、阿拉伯糖,相对摩尔比分别为3.3:1.1:0.29:37.2:0.76:39.7:0.2。PBMF具有较强还原力和羟自由基清除能力。PBMF对He La细胞的增殖具有显著地抑制作用,且呈剂量依赖性。PBMF诱导He La细胞周期停滞于G0/G1期(P<0.05)。上调caspase-3的相对表达量,诱导细胞早期凋亡和晚期凋亡。结论 PBMF能抑制人宫颈癌He La细胞增殖,并诱导细胞凋亡,其机制与caspase-3的活化有关。Objective To analyze the composition and antioxidant activity of the polysaccharides from black mulberry fruits（PBMF）, and explore the effects of PBMF on the apoptosis and proliferation of He La cells（human cervical carcinoma cell）. Methods PBMF were analyzed by UPLC. Reducing power and hydroxyl radical scavenging activities of PBMF were detected by antioxidant tests in vitro. MTT was used to detect the proliferation of He La cells. The cell morphology was observed by the inverted microscope. Cell apoptosis and cell cycle of He La cells were analyzed by flow cytometry（FCM） and Annexin V-FITC/PI double staining. Western blot was used to detect protein expression of procaspase-3 and caspase-3. Results PBMF were composed of seven monosaccharides as follows： mannose, rhamnose, glucuronic acid, glucose, xylose, galactose, and arabinose. The molar ratio was 3.3：1.1：0.29：37.2：0.76：39.7：0.2. PBMF had strong reducing power and hydroxyl radical scavenging activity. PBMF showed significant inhibitory effect on the growth of He La cells in a dose dependent manner. PBMF induced an arrest of He La cells in the G0/G1 phase（P〈0.05）. PBMF increased cell apoptosis rate and the expression of caspase-3. Conclusion PBMF could inhibit He La cells proliferation and induce apoptosis, which are correlated with the activation of caspase-3 expression.

关 键 词：药桑椹 多糖 超高效液相色谱 细胞凋亡 抗肿瘤 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学]