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40mg·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、40.0mg·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27mg;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;40mg·L^-1 FRIL组Raji细胞凋亡率明显低于20 mg·L^-1 FRIL组（P〈0.01）;与对照组比较,5.0-40.0mg·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。
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,20mg·L^-1and 40mg·L^-1FRIL groups;the cultured HL-60 cells were grouped into 0,2.5,5.0,10.0,20.0,and 40.0mg·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 containing4 subunits was obtained from Astragalus root,which had a high affinity for combiningα-D-manoside.The apoptotic rate of Raji cells in 40mg·L-1 FRIL group was lower than that in 20mg·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-1FRIL 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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2015年第1期105-109,共5页
Journal of Jilin University：Medicine Edition
基金
吉林省中医药管理局科技项目资助课题(2012-125)
