HGF拮抗TGF－β1诱导腱鞘成纤维细胞α-SMA及细胞外基质合成  

HGF depress the production of α-SMA and extracellular matrix induced by TGF-β1 in fibroblasts of the flexor tendon sheath

作　　者：姜大朋[1] 李昭铸[1] 张玉波[1] 韩福友[1] 管声扬[1] 蒋志涛[1] 

机构地区：[1]哈尔滨医科大学附属第二医院小儿外科,哈尔滨150086

出　　处：《中华显微外科杂志》2010年第4期297-300,共4页Chinese Journal of Microsurgery

基　　金：基金项目：国家自然基金青年基金资助项目（30901516）;哈尔滨医科大学附属第二医院博士启动基金（BS2009-02）

摘　　要：目的研究HGF能否阻抑TGF－β1诱导的腱鞘成纤维细胞α－SMA及细胞外基质过度合成。方法选取成年新西兰大白兔7只，体重3．75～4．00kg，无菌切取前肢中趾趾深屈肌腱，进行腱鞘成纤维细胞的分离和培养，待细胞生长成单层后，以胰蛋白酶消化传代。取第3代成纤维细胞用于实验，当细胞达到70％融合时，培养液中加入TGF-β1（5ng／ml）及HGF（10～40ng／ml）。培养72h后．利用Westem blot检测α-SMA表达；ELISA测定细胞I型胶原及纤维结合素的表达。结果TGF-β1能显著诱导α-SMA表达，半定量分析提示，TGF-β1作用后的成纤维细胞α—SMA表达量是对照组的1．8倍。随HGF的同时加入，α—SMA的表达则明显受抑制（P〈0．05），且随HGF浓度的升高其阻抑作用呈逐渐增强趋势。TGF-β1同样能诱导I型胶原及纤维结合素的表达（P〈0．01），而HGF则可以有效地阻抑其表达，其效应呈剂量依赖性（P〈0．05）。结论HGF可以有效阻抑TGF-β1诱导的腱鞘成纤维细胞α-SMA、I型胶原及纤维结合素的表达，这为利用HGF预防和治疗屈指肌腱损伤后牯连及瘢痕在细胞和分子水平提供了依据。Objective To examine the effectiveness of HGF in blocking TGF-β1 induced α-SMA and extracellular matrix production in fibroblasts of the flexor tendon sheath. Methods Seven adult male New Zealand white rabbits （3.75-4.00 kg） were used for this study. Both of their front feet were sterilised and the middle digit flexor digitorum profundus tendon equivalents were identified and isolated. These specimens were used to establish primary cell cultures. Sheath fibroblasts were obtained from rabbit flexor tendons. After the cells reached confluence, cells were detached with trypsin/ethylenediamine tetra-acetic acid. All experiments were performed using the cells at the third passage. At 70% confluence the medium was supplemented with 5 ng/ml of TGF-β1 along with increasing doses of HGF （10-40 ng/ml）. After 72 hours incubation, the productions of α-SMA were assayed by Western-Blot. The productions of collagen I and fibronectin in supernatants culture were examined using ELISA. Results Evaluation of protein expression revealed that TGF-β1 markedly induced α-SMA expression in cultured rabbit flexor tendon sheath fibroblasts. TGF-β1 treated fibroblasts expressed 1.8-fold more protein compared to non-treated fibroblasts （P 〈 0.05）. However, simultaneous incubation of HGF significantly abrogated TGF-β1 induced α-SMA expression in a dose-dependent manner （P 〈 0.05）. Treatment with TGF-β1 significantly stimulated collagen I and fibronectin production in flexor ten- don sheath fibroblasts （P 〈 0.01 ）. Remarkably, the addition of HGF reduced productions of all components induced by TGF-β1 in a dose-dependent manner （P 〈 0.05）. Conclusion HGF antagonizes TGF-β1 induced α-SMA, collagen I, and fibronectin production in flexor tendon sheath fibroblasts. The findings provide a cellular and molecular basis for HGF＇s acting as a therapeutic agent for adhesions in flexor tendons.

关 键 词：肝细胞生长因子 转化生长因子-β 屈指肌腱 成纤维细胞 α-平滑肌肌动蛋白 

分 类 号：R658.2[医药卫生—外科学]