6-磷酸果糖对肌腱细胞TGF-β及其受体表达的影响  

EFFECTS OF MANNOSE-6-PHOSPHATE ON TRANSFORMING GROWTH FACTOR β AND TRANSFORMING GROWTH FACTOR β RECEPTOR EXPRESSION OF FLEXOR TENDON CELLS

作　　者：杨瑞祥[1] 夏长所[1] 王秀英[1] 孙康[1] 杨选影[1] 田少奇[1] 洪光祥[2] 

机构地区：[1]青岛大学医学院附属医院骨科,山东青岛266003 [2]华中科技大学同济医学院附属协和医院手外科

出　　处：《中国修复重建外科杂志》2010年第1期64-68,共5页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的通过体外培养兔肌腱的腱鞘、腱外膜和腱内膜细胞,观察6-磷酸果糖对3种细胞TGF-β及其受体、TGF-β1及TGF-β1mRNA表达的影响,探讨6-磷酸果糖在肌腱愈合粘连防治中的作用机制提供实验依据。方法成年新西兰大白兔8只,雌雄不限,体重4.0～4.5kg。取兔前肢趾深屈肌腱分离培养腱鞘、腱外膜和腱内膜细胞。将每种细胞随机分为2组,实验组加入6-磷酸果糖培养,对照组不加入6-磷酸果糖。分别于培养3d及24h后采用酶联免疫吸附实验定量检测TGF-β及其受体的表达,原位杂交检测TGF-β1mRNA的表达;免疫组织化学染色观测TGF-β1的表达。结果酶联免疫吸附实验显示实验组各细胞TGF-β及其受体的表达均较对照组下降,差异有统计学意义(P<0.05)。TGF-β1表达平均降低程度从大到小依次为腱鞘细胞(36.1%)、腱内膜细胞(31.2%)、腱外膜细胞(31.0%),TGF-β2依次为腱鞘细胞(37.9%)、腱外膜细胞(32.1%)、腱内膜细胞(27.0%),TGF-β3依次为腱内膜细胞(42.5%)、腱鞘细胞(41.2%)及腱外膜细胞(33.3%)。TGF-β受体1、2表达平均降低程度从大到小依次为腱外膜细胞(29.9%、26.2%),腱内膜细胞(27.8%、23.5%),腱鞘细胞(23.1%、20.0%);TGF-β受体3依次为腱内膜细胞(26.1%)、腱外膜细胞(19.2%)、腱鞘细胞(15.8%)。实验组各细胞TGF-β1mRNA阳性表达率及细胞内TGF-β1mRNA表达强度均较对照组明显降低,比较差异有统计学意义(P<0.05)。免疫组织化学染色观察显示实验组各细胞TGF-β1表达均较对照组明显降低。结论6-磷酸果糖能显著降低肌腱细胞的TGF-β及其受体、TGF-β1及TGF-β1mRNA的表达,为肌腱愈合过程中调节TGF-β水平提供了一种新途径。Objective By culturing tendon sheath fibroblasts,epitenon tenocytes and endotenon tenocytes of rabbits’tendon in vitro,to study the effects of mannose-6-phosphate on transforming growth factorβ（TGF-β）peptide and receptor expression,and to provide the experimental basis for preventing the tendon healing adhesion by mannose- 6-phosphate.Methods Eight adult New Zealand white rabbits,regardless of their gender and weighing 4.0-4.5 kg,were selected.Tendon sheath fibroblasts,epitenon tenocytes,and endotenon tenocytes were isolated from rabbit flexor tendon and cultured separately.All 3 cells were divided into 2 groups at random after cells were adjusted to a concentration of 4×104 per well and 1×104/mL.The first was the control group without supplementation.The experimental group was supplemented with mannose-6-phosphate.The expressions of TGF-βand TGF-βreceptor were quantified with enzyme-linked immunosorbent assay.The expression of TGF-β 1 mRNA was also assessed with in situ hybridization and the expression of TGF-β 1 was assessed with immunohistochemistry.Results The expressions of TGF-βand TGF-βreceptor in experimental group were significantly lower than that in control group（P0.05）.The expression levels of TGF-β 1 and TGF-β 2 decreased in descending order of tendon sheath fibroblasts（36.1%,37.9%）,epitenon tenocytes（31.0%,32.1%）,and endotenon tenocytes（31.2%,27.0%）.The expression levels of TGF-β 3 decreased in descending order of endotenon tenocytes（42.5%）,tendon sheath fibroblasts（41.2%）, and epitenon tenocytes（33.3%）.The expression levels of TGF-βreceptor 1 and TGF-βreceptor 2 decreased in descending order of epitenon tenocytes（29.9%,26.2%）,endotenon tenocytes（27.8%,23.5%）,and tendon sheath fibroblasts（23.1%,20.0%）. The expression levels of TGF-βreceptor 3 decreased in descending order of endotenon tenocytes（26.1%）,epitenon tenocytes （19.2%）,and tendon sheath fibroblasts（15.8%）.In experimental group,the positive expression of TGF-β 1 mRNA

关 键 词：肌腱细胞 6-磷酸果糖 TGF-Β TGF-Β受体 兔 

分 类 号：R341[医药卫生—基础医学]