改性柑橘果胶对滑膜成纤维细胞的影响  

Effect of modified citrus pectin on synovial fibroblasts

作　　者：陈雅珍 粟丹宁 郑嘉诺 何嘉越 段瑞平[1] 杜博[1] 李学敏[1] 刘玲蓉[1] Chen Yazhen;Su Danning;Zheng Jianuo;He Jiayue;Duan Ruiping;Du Bo;Li Xuemin;Liu Lingrong(Institute of Biomedical Engineering,Chinese Academy of Medical Sciences&Peking Union Medical College,Tianjin 300192,China)

机构地区：[1]中国医学科学院,北京协和医学院生物医学工程研究所,天津300192

出　　处：《国际生物医学工程杂志》2023年第2期97-103,共7页International Journal of Biomedical Engineering

基　　金：国家自然科学基金面上项目(81972043)。

摘　　要：目的研究改性柑橘果胶(MCP)对滑膜成纤维细胞(SF)及半乳糖凝集素-3(Gal-3)诱导处理后SF细胞的代谢活性及相关基因表达情况的影响。方法分离、培养兔SF细胞,分别用不同质量浓度(0、250、500、750 mg/L)MCP的培养液处理SF细胞,同时用10µg/ml Gal-3诱导24 h后进一步进行MCP处理SF细胞;分别于处理的第1、3、5天,通过CCK-8检测SF细胞的增殖活性;利用实时荧光定量PCR检测SF细胞的转化生长因子-β1(TGF-β1)、Ⅰ型胶原(COL1A2)及Gal-3的mRNA表达情况,并通过免疫荧光染色法考察SF细胞的COL1A2合成情况。结果在第1、3、5天,不同质量浓度MCP均对SF细胞增殖有抑制作用(均P<0.05)。与对照组相比,不同浓度的MCP可以引起不同的基因表达,高浓度的MCP上调了SF细胞的TGF-β1、COL1A2和Gal-3的mRNA表达;同时,MCP对SF细胞的COL1A2蛋白合成无影响。Gal-3诱导后,MCP处理可以下调TGF-β1和COL1A2的mRNA表达,且可以显著降低COL1A2蛋白的合成;尤其是500 mg/L MCP可以明显降低TGF-β1、COL1A2和Gal-3的mRNA表达。结论MCP具有抑制SF细胞过度增殖、调控SF细胞基因表达的作用。Objective To study the effects of modified citrus pectin(MCP)on the viability and gene expressions of synovial fibroblasts(SF)as well as SF treated by galectin-3(Gal-3).Methods Rabbit SF was isolated and cultured in vitro.Then SF was treated with different concentrations of MCP(0,250,500,and 750 mg/L).In addition,SF was further treated with the same different concentrations of MCP after treatment with 10µg/ml Gal-3 for 24 h.The viability of SF was detected by CCK-8 on the first,third,and fifth day after treatment.The mRNA expression of transforming growth factor-β1(TGF-β1),type I collagen(COL1A2),and Gal-3 in SF was detected by real-time quantitative PCR.The synthesis of type I collagen in SF was investigated by immunofluorescence staining.Results MCP,especially at a concentration of 500 mg/L can inhibit the proliferation of SF significantly(all P<0.05)on the first,third,and fifth day after treatment.Compared with the control group,MCP at different concentrations induced different gene expression profiles.In particular,MCP at high concentrations can upregulate the expression of TGF-β1,COL1A2 and Gal-3 in SF.However,MCP shows no significant effect on the synthesis of type I collagen in SF.MCP can down-regulate the expression of TGF-β1,COL1A2,and significantly reduce the synthesis of type I collagen in SF after Gal-3 treatment.Particularly,the effect of MCP at a concentration of 500 mg/L on inhibiting the expression of TGF-β1,COL1A2,and Gal-3 in SF is significant.Conclusions MCP can inhibit the excessive proliferation of SF and regulate gene expression in SF.

关 键 词：改性柑橘果胶 骨关节炎 滑膜成纤维细胞 滑膜纤维化 半乳糖凝集素-3 

分 类 号：R684.3[医药卫生—骨科学]