黄芪多糖和黄芪总甙调控LX-2细胞因子分泌  

Astragalus polysaccharides and astragalosides regulate cytokine secretion in LX-2 cell line

作　　者：李淑萍[1] 许晓燕[2] 孙箴[2] 周林福[2] 羊正纲[1] 陈峰[1] 朱海红[1] 陈智[1] 

机构地区：[1]浙江大学医学院附属第一医院传染病研究所,浙江杭州310003 [2]浙江大学医学院医学生物技术实验室,浙江杭州310058

出　　处：《浙江大学学报（医学版）》2007年第6期543-548,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省中医药管理局科技计划(2006C075)资助项目

摘　　要：目的:用黄芪多糖和黄芪总甙对LX-2细胞分泌的与纤维化相关的主要细胞因子进行研究,以探讨黄芪抗纤维化的确切分子机制。方法:不同浓度的黄芪多糖和黄芪甙作用于LX-2细胞,以Real Time Cell Electronics Sensing技术实时检测细胞增殖状态;分别再以25μg/ml和300μg/ml的黄芪多糖和黄芪总甙作用LX-2细胞24 h,提取细胞mRNA,以荧光定量PCR测定转化生长因子1(TGF-β1)、肝细胞生长因子(HGF)、基质金属蛋白酶2(MMP2)及基质金属蛋白酶9(MMP9)mRNA的变化。结果:300μg/ml的黄芪多糖可抑制LX-2的增殖(P<0.05,94.7 h),但不同浓度的黄芪总甙对LX-2细胞均有促进增殖的作用(P<0.05);25μg/ml的黄芪多糖和黄芪总甙在TGF-β1促进分泌的同时,也上调其它抗纤维化因子,其中黄芪多糖对MMP2的上调(104.9倍)、黄芪总甙对IL-10的上调(550.65倍)最为显著;300μg/ml的黄芪多糖可抑制TGF-β1及HGF、MMP9、IL-10的表达,但仍然上调MMP2的表达,300μg/ml的黄芪总甙也可抑制TGF-β1的表达,但对抗纤维化因子MMP2、MMP9及IL-10仍有明显的上调作用。结论:低浓度(25μg/ml)和高浓度(300μg/ml)的黄芪多糖和黄芪总甙可通过不同的分子机制调控肝星状细胞因子的分泌而发挥抗纤维化作用;黄芪多糖主要以促进MMP2分泌为主,而黄芪总甙则主要通过上调MMP2、MMP9及IL-10的分泌而发挥作用。高剂量的黄芪多糖和总甙抗纤维化效果优于低剂量组。Objective： To investigate the anti-fibrosis mechanism of radix astragali through the observation on regulating cytokine secretion by astragalosides and astragalus polysaccharides in hepatic stellate cell line LX-2. Methods： Astragalus polysaccharides and astragalosides at concentration of 25 μg/ml and 300 μg/ml were added to LX-2 cell culture. After 24 h culture total RNA contents were extracted and TGF-β1,HGF,MMP2 and MMP9 mRNA were measured by real time fluorescence quantification PCR technique. The cell growth curves were detected by Real Time Cell Electronics Sensing. Results： 300 μg/ml of astragalus polysaccharides suppressed LX -2 cell proliferation （P〈0. 05, in 94. 7 h）, while astragalosides induced an significant cell proliferation （P 〈 0. 05 ） both at lower and higher concentration. 25 μg/ml astragalus polysaccharides and astragalosides induced TGF-β1 expression. For other cytokines, astragalus polysaccharides induced a 104.9 fold higher expression of MMP2,while astragalosides induced a 550. 65 fold higher expression of IL-10. Astragalus polysaccharides at 300 μg/ml concentration exhibited an inhibition effect on TGF-β1,HGF,MMP9 and IL-10 mRNA expression,while up -regulated MMP2 mRNA expression. Astragalosides at 300 μg/ml concentration inhibited TGF-β1 mRNA expression,while up-regulated MMP2, MMP9 and IL-10 mRNA expression. Conclusions： The anti-fibrosis function of astragalus polysaccharides might be associated with up-regulation of MMP2 expression, while that of astragalosides with up-regulation of MMP2, MMP9 and IL-10 expression.

关 键 词：黄芪多糖/投药和剂量 黄芪甙/投药和剂量 细胞因子类 LX-2细胞 细胞因子 肝纤维化 

分 类 号：R285.5[医药卫生—中药学]