天然水蛭素对肌腱成纤维细胞抑制作用的体外研究  

Research on the Inhibitory Effect on Tendon Fibroblasts of Natural Hirudin in Vitro

作　　者：曾林如 汤样华[1] 王楠[1] ZENG Linru;TANG Yanghua;WANG Nan(Xiaoshan TCM Hospital,Hangzhou 311201,China)

机构地区：[1]杭州市萧山区中医院,浙江杭州311201

出　　处：《中国医学创新》2018年第5期9-12,共4页Medical Innovation of China

基　　金：浙江省中医药科技计划项目(2015ZA175)

摘　　要：目的:通过体外实验研究天然水蛭素对兔肌腱成纤维细胞的抑制作用及三种相关细胞因子表达水平的变化,揭示天然水蛭素预防肌腱粘连的作用及相关途径。方法:体外分离培养兔肌腱成纤维细胞,采取MTT法检测不同浓度天然水蛭素对兔肌腱成纤维细胞增殖活性的影响;运用ELISA法测定兔肌腱成纤维细胞转化生长因子β1(TGF-β1)、碱性成纤维细胞生长因子(b FGF)和基质金属蛋白酶2(MMP-2)的表达含量。结果:与未加水蛭素组(0 U/m L)比较,天然水蛭素干预组(1.50、3.00、6.00 U/m L)能够显著抑制兔肌腱成纤维细胞生长;并正向调节b FGF与MMP-2含量表达,负向调节TGF-β1含量表达(P<0.05或<0.01)。结论:天然水蛭素能够显著抑制体外培养的兔肌腱成纤维细胞的增殖,且具有一定的浓度依赖性;通过降低兔肌腱成纤维细胞TGF-β1的表达并相应提高b FGF与MMP-2的表达可能是其基于TGF-β1/Smads信号通路发挥抗纤维化作用的一种有效途径。Objective:To reveal the function and pathway of preventing tendon adhesion of hirudin,we investigate the anti-fibrosis effect of hirudin and the expression of three kinds of related cytokines in rabbit tendon fibroblasts in vitro.Method:Firstly,culturing rabbit tendon fibroblasts in vitro,and then observing the morphological changes of each group.Nextly,proliferative activity of fibroblasts was detected by MTT method in different concentrations of hirudin.Finally,the expression levels of transforming growth factorβ1(TGF-β1),basic fibroblast growth factor(bFGF)and matrix metalloproteinase 2(MMP-2)in rabbit tendon fibroblasts were determinated by using the method of ELISA.Result:Hirudin intervention group(1.50,3.00,6.00 U/mL)could inhibit the growth of rabbit tendon fibroblasts compared with 0 U/mL group,and increase the expression of MMP-2 and bFGF,while decrease the expression of TGF-β1(P<0.05 or<0.01).Conclusion:Hirudin can inhibit the proliferative activity of rabbit tendon fibroblasts with concentration dependent in vitro.By increasing the expression of bFGF and MMP-2 and reducing the expressing of TGF-β1 of rabbit tendon fibroblasts may be an effective pathway of anti-fibrosis in hirudin based on TGF-β1/Smads pathway.

关 键 词：水蛭素 成纤维细胞 肌腱粘连 TGF-β1 BFGF MMP-2 

分 类 号：R285.5[医药卫生—中药学]