重组人内抑素对人瘢痕疙瘩成纤维细胞形态及增殖的影响  

Effects of rhEndostatin on the morphology and proliferation of human keloid-derived fibroblasts in vitro

作　　者：吕玉琳[1] 张晓明[1] 黄学应[1] 邓雪飞[1] 李小静[2] 左宗宝[3] 

机构地区：[1]安徽医科大学人体解剖学教研室 [2]安徽医科大学第一附属医院整形外科 [3]合肥市第一人民医院整形外科,合肥230032

出　　处：《解剖学报》2012年第3期347-351,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(81171819);安徽医科大学校级科研基金资助项目(2010xkj007)

摘　　要：目的观察重组人内抑素(rhEndostatin)对体外培养的人瘢痕疙瘩成纤维细胞(KFs)增殖的影响。方法体外培养及鉴定人KFs;应用四甲基偶氮唑盐(MTT)法分别检测不同浓度rhEndostatin(6.25×10-6、1.25×10-5、2.50×10-5、5.00×10-5、1.00×10-4mg/L)作用24h、48h、72h后对KFs增殖的影响,并同时观察其形态变化。结果组织块接种7～8d后,其周边可见少许梭形细胞,继之细胞逐渐增多并以组织块为中心呈放射状生长;传2代培养的细胞单层排列,形态均一,呈长梭状,折光性好。细胞波形蛋白免疫化学染色显示胞质均呈棕黄色。rhEndostatin(6.25×10-6mg/L浓度组除外)能明显抑制KFs的增殖(P<0.05),抑制效应与rhEndostatin浓度和作用时间呈一定的依赖关系;其中,5.00×10-5mg/L和1.00×10-4mg/L组rhEndostatin作用48 h后,细胞生长缓慢,体积变小,数目减少,排列紊乱。结论重组人内抑素对人KFs增殖具有抑制作用。Objective To analyze the inhibitory effect of recombinant human endostatin（rhEndostatin） on human keloid-derived fibroblasts（KFs） in vitro. Methods The human＇ s KFs were cultured and identified in vitro. KFs were incubated with various concentrations of rhEndostatin solution （6. 25 ×10 -4, 1. 25× 10 -5, 2. 50 × 10 -5, 5.00 × 10 -5, 1.00×10 -4 mg/L）. With the 3- （4,5-dimethylthiazol-2-yl） -2, 5-diphenny-lterazolium bromide （ MTT ） colorimetric assay the effects of rhEndostatin on the proliferation of cultured KFs were studied, and morphological changes of the KFs were also observed at different time points（24, 48, 72 hours）. Results KFs from the keloid tissue started to appear on days 7-8 after it were planted, the number of the cells gradually increased and the growth of the cells showed a radial patten centred at the tissue-centric. Second subculture KFs had a single-layer arrangement, were uniform, showed long fusiform, and had good refractive index. Mouse anti-Vimentin was positive. Buffy positive reaction product was observed in the cytoplasm. Cells growth inhibition occurred at 1.25 ×10-5 mg/L or higher concentration in a dose-dependent manner and maximum inhibition occurred at 48 hours. After incubation with rhEndostatin（5.00 × 10-5 mg/L and 1.00 × 10-4 mg/L） for 48 hours ,observation under the microscope showed the cells slowed growth, reduced volume, reduced the number of cells and disorders. Conclusion A certain concentration of rhEndostatin may inhibit the proliferation of KFs.

关 键 词：内抑素 瘢痕疙瘩 成纤维细胞 增殖 四甲基偶氮唑盐法 

分 类 号：R619.6[医药卫生—外科学]