人参多糖对大鼠关节软骨细胞糖胺聚糖合成的影响及其机制
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- 英文论文题名：Effects and mechanism of Giseng polysaccharides on glycosaminoglycan synthesisof rat chondrocytesin vitro
- 论文作者：李景 ; 兰昱 ; 宋金春
- 英文论文作者：Li Jing ; Lan Yu ; Song jinchun ; Department of Pharmacy ; Remin Hospital of Wuhan University
- 年：2016
- 作者机构：武汉大学人民医院药学部;
- 论文关键词：人参多糖 ; 骨关节炎 ; 糖胺聚糖 ; 木糖基转移酶-1 ; 半乳糖基转移酶-Ⅰ ; 半乳糖基转移酶-Ⅱ ; 葡萄糖醛酸转移酶-Ⅰ
- 英文论文关键词：Giseng polysaccharides ; osteoarthritis ; glycosaminoglycan ; XylT-1 ; GalT-Ⅰ ; GalT-Ⅱ ; GlcAT-Ⅰ
- 会议召开时间：2016-09-27
- 会议录名称：中国药学会第十三届青年药学科研成果交流会论文集
- 语种：中文
- 分类号：R285.5
- 学会代码：YYWS
- 会议名称：中国药学会第十三届青年药学科研成果交流会
- 会议地点：中国江西南昌
- 主办单位：中国药学会
- 学会名称：中国药学会
- 页数：6
- 文件大小：1041k
- 原文格式：O
- 会议级别：全国
摘要
目的:探讨人参多糖对大鼠关节软骨细胞糖胺聚糖(Glycosaminoglycan,GAG)合成的影响及其作用机制。方法:分离原代大鼠关节软骨细胞进行传代培养并加入白介素-1β造模,分别将2、10、50μg/mL的人参多糖作用于该软骨细胞48h。检测细胞增殖、GAG含量和GAG合成酶木糖基转移酶-1(xylosyltransferase-1,XylT-Ⅰ)、半乳糖基转移酶-Ⅰ(galactosyltransferaseⅠ,Ga1T-Ⅰ)、半乳糖基转移酶-Ⅱ(galactosyltransferaseⅡ,Ga1T-Ⅱ)和葡萄糖醛酸转移酶-Ⅰ(galactose-β-1,3-glucuronosyltransferaseⅠ,GlcAT-Ⅰ)mRNA的含量和基质降解酶基质金属蛋白酶-3(Matrix metalloproteinases-3,MMP-3)、聚集蛋白聚糖酶-1(aggrecanase-1)和聚集蛋白聚糖酶-2(aggrecanase-2)mRNA的含量。结果:各浓度人参多糖对大鼠关节软骨细胞增殖无影响;10、50μg/mL GPS可增加软骨细胞GAG含量;GPS可剂量依赖性增加Xy1T-1、GalT-Ⅰ、Ga1T-Ⅱ和G1cAT-ImRNA的表达;IL-1β可显著增加软骨细胞蛋白多糖降解酶表达,GPS对蛋白多糖降解酶表达无抑制作用。结论:GPS可促进IL-1β下调的大鼠软骨细胞GAG合成,其机制可能与促进GAG合成酶表达增加而不是抑制基质降解酶表达有关。
Objective To investigate the effects and mechanism of Giseng polysaccharides(GPS) on glycosaminoglycan synthesisof rat chondrocytes in vitro.Methods:Chondrocytes isolated from rat'sknee joint cartilage were cultured and passaged.After monolayers were established,chondrocytes induced by 20 ng /ml IL-1βwere treated with GPS of 2,10,50 ng/ml respectively for 48 h.The effects on chondrocytes were analyzed using MTT assay for chondrocyteproliferation and 1,9-dimethylmethylene blue assay for GAG content.mRNA expression of xylosyltransferase-1(XylT-Ⅰ),galactosyltransferase-Ⅰ(GalT-Ⅰ),galactosyltransferase Ⅱ(GalT-Ⅱ),galactose-β-1,3-glucuronosyltransferase Ⅰ(GlcAT-Ⅰ) and mRNA expression of matrix-degrading enzymes Matrix metalloproteinases-3(MMP-3),aggrecanase-land aggrecanase-2.Results:①GPS had no toxic or proliferatingeffect on chondrocytes in all observed concentrations.GPS of 10 ng /ml and 50 ng /ml up-regulated GAG synthesis(P<0.01);②GPS enhanced the expression of XylT-1,GalT-Ⅰ,GalT-Ⅱ and GlcAT-Ⅰ in a dose-dependent manner(P<0.05);③GPS did not inhibit the mRNA expression of matrix-degrading enzymes MMP-3,aggrecanase-land aggrecanase-2enhanced by 1L-1β.Conclusion:GPS can improve GAG synthesis of 1L-1β-stimulated chondrocytes in vitro,which is due to the promotion of expression of GAG sythesis enzymes but not the inhibition of the expression of matrix-degrading enzymes.
Objective To investigate the effects and mechanism of Giseng polysaccharides(GPS) on glycosaminoglycan synthesisof rat chondrocytes in vitro.Methods:Chondrocytes isolated from rat'sknee joint cartilage were cultured and passaged.After monolayers were established,chondrocytes induced by 20 ng /ml IL-1βwere treated with GPS of 2,10,50 ng/ml respectively for 48 h.The effects on chondrocytes were analyzed using MTT assay for chondrocyteproliferation and 1,9-dimethylmethylene blue assay for GAG content.mRNA expression of xylosyltransferase-1(XylT-Ⅰ),galactosyltransferase-Ⅰ(GalT-Ⅰ),galactosyltransferase Ⅱ(GalT-Ⅱ),galactose-β-1,3-glucuronosyltransferase Ⅰ(GlcAT-Ⅰ) and mRNA expression of matrix-degrading enzymes Matrix metalloproteinases-3(MMP-3),aggrecanase-land aggrecanase-2.Results:①GPS had no toxic or proliferatingeffect on chondrocytes in all observed concentrations.GPS of 10 ng /ml and 50 ng /ml up-regulated GAG synthesis(P<0.01);②GPS enhanced the expression of XylT-1,GalT-Ⅰ,GalT-Ⅱ and GlcAT-Ⅰ in a dose-dependent manner(P<0.05);③GPS did not inhibit the mRNA expression of matrix-degrading enzymes MMP-3,aggrecanase-land aggrecanase-2enhanced by 1L-1β.Conclusion:GPS can improve GAG synthesis of 1L-1β-stimulated chondrocytes in vitro,which is due to the promotion of expression of GAG sythesis enzymes but not the inhibition of the expression of matrix-degrading enzymes.
引文
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[2]Hashiomto S,Ochs RL,Rosen F,et al.Chondrocyte derived apoptotic bodies and calcification of articular cartilage[J].Proc Natl Acad Sci USA,1998,95:3094-3099.
[3]Uyama.T,et al.Biosynthesis of glycosaminoglycans and proteoglycans[J].Comprehensive Glycoscience,2007,3.05:79-104.
[4]Venkatesan N,Barre L,Bourhim M,Magdalou J,et al.Xylosyltransferase-I regulates glycosaminoglycan synthesis during the pathogenic process of human osteoarthritis[J].PLoS One.2012,7(3):e34020.
[5]谭杨,李景,汪晖,等.人参多糖抗大鼠骨关节炎及其机制研究[J].中药药理与临床,2013,29,3:91-93
[6]杨国志,李振武,赵瑞强,等.人参多糖、黄芪多糖对大鼠骨关节炎细胞模型葡糖氨基聚糖合成的影响及其可能机制[J].中国生化药物杂志,2014,08,34:57-64
[7]Lim T,Na K,Choi E,et al.Immunomodulating Activities of Polysaccharides isolated from Panax ginseng[J].J Medicinal Food,2004,7(1):1.
[8]Kltagawa H,UJIKAWA M,Sugahara K.Developmental changes in serum UDP-G1cA:chondrolt in gulcuronyltransferase activity[J].J Biol Chem,1996,271(12):6583-6585.
[9]Kiani C,Chen L,Wu YJ,et al.Structure and function of aggrecan[J].Cell Res.2002,12(1):19-32.
[10]Sandrine Gulberti,Virginie Lattard,Jean-Claude Jacquinet,et al.Phosphorylation and sulfation of oligosaccharide substrates critically influence the activity of human betal,4-galactosyltransferase 7(Ga1T-I)and betal,3-glucuronosyltransferase I(G1cAT-I)involved in the biosynthesis of the glycosaminoglycan-protein linkage region of proteoglycans[J].J Biol Chem,2005,280(2):1417-1425.
[11]Thollas B,Jacquinet JC.Synthesis of various sulfoforms of the trisaccharide beta-d-GlcpA-(1->3)-beta-d-Galp-(1->3)-beta-d-Galp-(1->OMP)as probes for the study of the biosynthesis and sorting of proteoglycans[J].Org Biomol Chem,2004,2(3):434-442.
[12]Venkatesan N,et al.Stimulation of proteoglycan synthesis by glucuronosyltransferase-I gene delivery:a strategy to promote cartilage repair[J].Proc Natl Acad Sci U S A,2004,101(52):18087-18092.
[13]Li,J P,et al.Glucuronyl C5-epimerase an enzyme converting glucuronic acid to iduronic acid in heparan sulfate/heparin biosynthesis[J].Prog Mol Biol Transl Sci,2010,93:59-78.