淫羊藿苷对藻酸盐3D体外培养下小鼠软骨细胞周期蛋白、β-catenin表达的影响  

Effects of icariin on expression of Cyclins andβ-catenin in mouse chondrocytes under alginate 3D culture in vitro

作　　者：王鹏珍 熊喜峰 秦胜男 张金丽 李爱国 WANG Pengzhen;XIONG Xifeng;QIN Shengnan;ZHANG Jinli;LI Aiguo(Guangzhou Red Cross Hospital,Guangzhou 510220,China)

机构地区：[1]广州市红十字会医院广州市创伤外科研究所,广州510000

出　　处：《山东医药》2020年第17期30-33,共4页Shandong Medical Journal

基　　金：国家自然科学基金青年基金项目(81902802);广州市中医药和中西医结合科技项目(20192A010009);广州市卫生健康科技一般引导项目(20181A010017,20191A011018)。

摘　　要：目的观察淫羊藿苷(ICA)对藻酸盐3D体外培养下小鼠软骨细胞周期蛋白Cyclin D1、Cyclin E及β-catenin表达的影响。方法分离培养得到新生原代小鼠关节的软骨细胞,体外构建藻酸盐3D复合体并分为对照组和ICA组,分别于含10μmol/L ICA、不含ICA的DMEM完全培养基培养14 d;采用阿利新蓝组织学染色观察软骨细胞活性和酸性黏多糖合成情况,分别采用免疫组化、荧光定量PCR法检测Cyclin D1、Cyclin E、β-catenin蛋白和基因。结果ICA组成簇状排列的软骨细胞明显多于对照组,阿利新蓝染色强度明显深于对照组,并可见大量酸性黏多糖集聚于软骨陷窝及周边。与对照组比较,ICA组软骨细胞中Cyclin D1、Cyclin E、β-catenin蛋白及基因表达均增加(P均<0.05)。结论ICA通过上调Cyclin D1、Cyclin E、β-catenin表达,以增加小鼠关节软骨细胞活性和酸性黏多糖合成。Objective To explore the effects of icariin(ICA)on expression ofβ-catenin,Cyclin D1 and Cyclin E in articular chondrocytes under alginate 3D culture in vitro.Methods The primary articular chondrocytes were isolated and obtained from neonatal mice.The chondrocyte samlpes were obtained by alginate 3D culture for 14 days in vitro,and were divided into the control group and ICA group(n=6),which were cultured in the DMEM complete medium containing 10μmol/L ICA and no ICA for 14 days,respectively.We used aricin blue histological staining to observe the activity of chondrocytes and the synthesis of acid mucopolysaccharide.The expression and gene ofβ-catenin,Cyclin D1 and cyclin E were detected by immunohistochemistry and real-time PCR,respectively.Results When the chondrocytes were cultured for 14 days,ICA could significantly increase the activity of chondrocytes and the synthesis of acidic mucopolysaccharides;ICA promoted the expression ofβ-catenin,Cyclin D1 and Cyclin E in protein and mRNA levels in chondrocytes,and the differences were statistically significant as compared with those of the control group(all P<0.05).Conclusions ICA can increase the activity of mouse articular chondrocytes and promote the synthesis of acidic mucopolysaccharides by up-regulating the expression of Cyclin D1 and Cyclin E,andβ-catenin.

关 键 词：淫羊藿苷 软骨细胞 藻酸盐 周期蛋白D1 周期蛋白E β-catenin蛋白 

分 类 号：R285[医药卫生—中药学]