黄芪多糖对体外培养大鼠软骨细胞内糖胺多糖合成的影响及其机制

目的:研究黄芪多糖对大鼠退变软骨细胞糖胺多糖(GAG)合成的影响。方法:分离大鼠关节软骨细胞传代培养并加入IL-1β造模,分别将100,10,1,0.1 mg/L的黄芪多糖作用于该软骨细胞,以4.5 mg/L的硫酸氨基葡萄糖作为阳性对照。其后检测细胞增殖、细胞内GAG含量、细胞内葡萄糖醛酸转移酶Ⅰ(GlcAT-1)mRNA含量和透射电镜观察细胞形态学改变。结果:各浓度的黄芪多糖均对大鼠软骨细胞增殖率无影响。100 mg/L的黄芪多糖可以增加软骨细胞内GAG的合成(P<0.01);1,10,100 mg/L的黄芪多糖能够增加GlcAT-1 mRNA的表达。结论:黄芪多糖能逆转IL-1β对体外培养的大鼠软骨细胞GAG合成的抑制作用,其主要机制可能与促进GlcAT-1的表达有关。

Effects and Mechanism of Astragalus Polysaccharides on Glycosaminoglycan Synthesis of Rat Osteoarthritic Chondrocytes in Vitro

Objective: To investigate the effects and mechanism of astragalus polysaccharides(APS) on rat osteoarthritic chondrocytes' glycosaminoglycan(GAG) synthesis in vitro.Methods: Chondrocytes were isolated from rat's knee and shoulder joint cartilage for culturing and passage.After monolayers were established,chondrocytes induced by IL-1β were treated with APS of 0.1,1,10 and 100 mg/L respectively.The effects on chondrocytes were analyzed using 3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2Htetrazoliuminner salt(MTS) assay for chondrocyte proliferation and 1,9-dimethylmethylene blue assay with purified shark chondroitin sulphate(CS) as a standard for GAG content.The mRNA expression of galactose-β-1,3-glucuronosyltransferaseⅠ(GlcAT-1) by rat chondrocytes was analyzed using a quantitative multistandard RT-PCR.Results: ① APS had no toxic or proliferating effect on chondrocytes in all observed concentrations.② APS at the concentration of 100 mg/L up-regulated GAG synthesis(P<0.01),③ APS at the concentration of 1,10 and 100 mg/L enhanced the expression of GlcAT-1.Conclusion: Astragalus polysaccharides can reverse the depression of GAG synthesis of rat chondrocytes induced by IL-1β in vitro through enhancing the expression of GlcAT-1 probably.