机构地区:[1]第三军医大学西南医院全军烧伤研究所,创伤,烧伤与复合伤国家重点实验室 [2]兰州军区乌鲁木齐总医院烧伤科,730050
出 处:《中华烧伤杂志》2006年第1期33-37,共5页Chinese Journal of Burns
基 金:国家重点基础研究发展规划资助项目(G1999054205)
摘 要:目的观察部分创面外用抗菌药物与成纤维细胞生长因子(FGF)2、表皮生长因子 (EGF)、重组人生长激素(rhGH)对成纤维细胞生物学特性的影响。方法体外培养成纤维细胞, 按所加药物不同分为对照组(常规培养)、丁胺卡那霉素(0.021、0.210、2.100 mg/L)组、庆大霉素(5、 50、500 mg/L)组、氯霉素(0.01、0.10、1.00 mg/L)组、磺胺米隆(5、10 g/L)组、FGF2(2 400 U/ml)组、 EGF(2 000 U/ml)组及rhGH(0.016、0.160、1.600 g/L)组。用噻唑蓝(MTT)法测定各组成纤维细胞增殖活性[吸光度(A)值],用流式细胞仪检测细胞周期,并于显微镜下观察细胞形态的变化。结果 (1)MTT法检测:与对照组A值0.455 3±0.021 7比较,各种剂量丁胺卡那霉素组、庆大霉素组、氯霉素组、磺胺米隆组成纤维细胞A值均明显降低(P<0.05或0.01),其中磺胺米隆(5、10 g/L)组降低最明显,分别为0.101 3±0.001 1、0.095 0±0.004 1(P<0.01)。FGF2组及0.016 g/L rhGH 组细胞A值明显高于对照组(P<0.05),而EGF组及0.160、1.600 g/L rhGH组A值与对照组接近 (P>0.05)。(2)细胞周期检测:对照组细胞增殖指数(PI)为(9.63±0.45)%,与之比较,0.210 mg/L丁胺卡那霉素组细胞PI值无明显变化(P>0.05),FGF2组、EGF组及0.016 g/L rhGH组PI值均明显升高,分别为(46.76±2.33)%、(42.30±1.41)%、(13.29±0.47)%(P<0.05或0.01)。 (3)形态学观察:对照组、EGF组及0.160、1.600 g/L rhGH组成纤维细胞数目较多,呈长条形或梭形, 轮廓不清,透明度高;丁胺卡那霉素组、庆大霉素组、氯霉素组、磺胺米隆组细胞数目较少,形态不规则,轮廓清晰,透明度低,细胞内多有颗粒样物质及空泡;FGF2组、0.016 g/L rhGH组细胞分布均匀、密集,呈长条形或梭形,核分裂相多见,轮廓不清,透明度高。结论不同创面外用药物对成纤维细胞生物学特性的影响各异,在创面修复过程中应选择合适的创面外用Objective To investigate the influence of some topically used antibiotics (amikacin, gentamicin, chloromycetin and sulfamylon), basic fibroblast growth factor (FGF2), epithelial growth factor (EGF) and recombinant human growth hormone (rhGH) on the growth of fibroblasts in vitro. Methods Fibroblasts were cultured and passaged. The cultured cells were then divided into control (routine culture of fibroblasts) , amikacin (amikacin in respective dose of 0. 021 , 0. 210, 2. 100 rag/L) , gentamicin ( in respective dose of 5, 50, 500 mg/L) , chloromycetin (in respective dose of 0.01, 0.10, 1.00 mg/L) , sulfamylon (in respective dose of 5, 10 g/L) , FGF2 (2 400U/ml) , EGF (2 000 U/ml) and rhGH (0. 016, 0. 160, 1.600 g/L) groups. After the above agents were added to the culture medium respectively, the proliferation of the cultured fibroblasts was determined with MTT method, and the result was expressed as A ( absorption) value. The cell cycle was determined with flow cytometry and the morphology of the cells was observed with inverted microscope. Results ( 1 ) MTT method : The A value of fibroblasts cultured with amikacin, gentamicin, chloromycetin and sulfamylon in various doses was obviously lower than that in control group (0.455 3±0.021 7, P 〈0.05 or0.01), and theAvalue ofsulfamylon group was the lowest in two doses (0.101 3±0.001 1 for5 g/L and0.095 0±0.004 1 for 10 g/L, P 〈0.01).On the other hand, the A value in FGF2 and rhGH group(0.016 g/L) was much higher than that in the control ( P 〈0.05). However,the A value in EGF (both doses) and rhGH groups(0. 160,1. 600 g/L) was close to that in control ( P 〉 0.05 ). (2) Cell cycle determination: The proliferation index (PI) of fibroblas ts cultured with amikacin in dose of 0.210 mg/L showed no difference compared to that in control (9.63 ±0.45)% , ( P 〉0.05). But the PI of fibroblasts cultured with FGF2, EGF and rhGF in dose of 0. 016 g/L was increas
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