吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (01): 105-109.doi: 10.13481/j.1671-587x.20150120

• 基础研究 • 上一篇    下一篇

黄芪中新型FLT3受体相互作用凝集素的生物学活性分析

段秀梅, 王银萍, 毕丽荣, 田庄   

  1. 吉林大学第一医院临床病理诊断中心, 吉林 长春 130021
  • 收稿日期:2014-07-18 发布日期:2015-01-30
  • 通讯作者: 段秀梅,教授,硕士研究生导师(Tel:0431-81875916,E-mail:xmduan@jlu.edu.cn) E-mail:xmduan@jlu.edu.cn
  • 作者简介:段秀梅(1967-),女,吉林省长春市人,教授,医学博士,主要从事肿瘤分子病理学方面的研究。
  • 基金资助:

    吉林省中医药管理局科技项目资助课题(2012-125)

Analysis on bioactivity for new-type FLT3 receptor interacting lectin from Astragalus root

DUAN Xiumei, WANG Yinping, BI Lirong, TIAN Zhang   

  1. Center of Clinical Pathological Diagnosis, First Hospital, Jilin University, Changchun 130021 China
  • Received:2014-07-18 Published:2015-01-30

摘要:

目的: 从黄芪中提取、纯化Fms样酪氨酸激酶3(FLT3)受体相互作用凝集素(FRIL),分析鉴定其生物学活性。方法: 采用亲和层析方法纯化黄芪水浸粗提物,用BCA蛋白测定法测定FRIL含量,糖抑制红细胞凝集实验鉴定FRIL与糖结合的生化特性;培养Raji细胞,分为对照组和20及40 mg·L-1FRIL组;培养FLT3受体高表达细胞株HL-60细胞,分别设置对照组和2.5、5.0、10.0、20.0、40.0 mg·L-1 FRIL组;流式细胞术和MTT法分别检测FRIL对Raji细胞和HL-60细胞的增殖作用。结果: 从黄芪中分离得到含有4个亚基的凝集素FRIL达到27 mg;FRIL主要与α-D-甘露糖有很高的亲和性,证实FRIL能与糖专一性结合具有凝集素活性;40 mg·L-1FRIL组Raji 细胞凋亡率明显低于20 mg·L-1FRIL组(P<0.01);与对照组比较, 5.0~40.0 mg·L-1FRIL组HL-60细胞体外增殖活性增加(P<0.05)。结论: 成功从黄芪中分离得到具有凝集素活性的FRIL蛋白,对表达FLT3受体的细胞具有刺激增殖作用。

关键词: 黄芪, FLT3受体相互作用凝集素, 细胞增殖

Abstract:

Objective To extract and purify the Fms-like tyrosine kinase-3(FLT3) receptor interacting lectin (FRIL) from Astragalus root, and to analyze and identify its bioactivity. Methods Affinity chromatography was used to purify the crude extraction of Astragalus root;the concentration of FRIL was determined using BCA assay;the property of FRIL combined with sugar was verified using inhibition lectin reaction;the Raji cells were cultured and divided into control, 20 mg·L-1and 40 mg·L-1FRIL groups;the cultured HL-60 cells were grouped into 0, 2.5, 5.0, 10.0, 20.0, and 40.0 mg·L-1 FRIL groups;the proliferation effect of FRIL on human Raji cells and HL-60 cells were measured using flow cytometry and MTT method. Results The 27 mg of FRIL containing 4 subunits was obtained from Astragalus root, which had a high affinity for combining α-D-manoside.The apoptotic rate of Raji cells in 40 mg·L-1 FRIL group was lower than that in 20 mg·L-1 FRIL group(P<0.01).Compared with control group, the proliferation activities of the HL-60 cells expressing FLT3 receptors in 5.0-40.0 mg·L-1 FRIL groups were increased (P<0.05). Conclusion The FRIL protein possessing lectin activity is successfully isolated from Astragalus root, which has the promotion effect on the growth of the cells expressing FLT3 receptor.

Key words: astragalus root, FLT3 receptor interacting lectin, cell proliferation

中图分类号: 

  • R284.1