不同分子质量当归多糖对大鼠正常肝细胞内hepcidin表达的影响

Inhibitory effects of different molecular weight polysaccharides from Angelica sinensis on hepcidin expression in normal rat hepatocytes in vitro

目的:研究不同分子质量当归多糖(ASP1、ASP2)对大鼠正常肝细胞hepcidin表达的直接抑制作用。方法:采用改良的"原位在体两步灌流法"分离出原代大鼠肝细胞;以2 IU.mL-1的重组人促红细胞生成素(rhEPO)作为阳性对照,实验组中2种分子质量当归多糖分别分为高、中、低剂量,在此基础上根据细胞培养时间的不同再进一步分组;ELISA检测细胞上清中hepcidin的累积分泌量。结果:rhEPO作用24 h后,肝细胞hepcidin的累积分泌量下降52.6%;大分子质量ASP1对hepcidin表达无明显影响;而小分子质量ASP2在连续作用48 h后产生显著的hepcidin抑制效应,且抑制强度呈剂量相关性。结论:当归多糖中的大分子质量段在体外不能直接影响大鼠正常肝细胞内hepcidin的表达含量,而小分子质量段可以直接作用于肝细胞抑制hepcidin表达。

OBJECTIVE To clarify the direct inhibitory effects of two kinds of ASP （ASP1 and ASP2） on hepcidin expres sion in vitro. METHODS Rat primary hepatocytes were isolated after two-step perfusion of liver performed in situ. Recombi nant human erythropoietin （rhEPO, 2 U.mL-1） was designed as positive control group, both of ASP1 and ASP2 treatment group included three different doses groups respectively, and then the groups were further divided into sub groups according to the incubating time. Enzyme-linked immunosorbent assay was used to determine the accumulated expression of hepcidin. RESULTS Hepcidin expression in hepatocytes decreased 52.6% after 24 h incubated with rhEPO. There was no significant differences in hepcidin expression of hepatocytes treated with high molecular weight ASP1. Howev4r, when the ceils were exposed to ASP2 （low molecular weight） with 48 h, the hepcidin contents decreased significantly （23.6%, 63% and 73.7%, respectively）, which showed the positive correlation between the effects and dosages. CONCLUSION The low molecular weight ASP could directly inhibit the expression of hepcidin in hepacytes while the high molecular weight fraction did not.