抗生素及bFGF,EGF,rhGH对人体外培养Hacat细胞生物学特性影响

Effect of some antibiotics,bFGF,EGF and rhGH on biological characteristics of Hacat cells in vitro

目的观察不同种类不同浓度的创面外用药物对Hacat(hum an adu lt sk in keratinocyte propagated under lowCa and elevated temperature)细胞生物学特性的影响。方法将Hacat细胞传代,经创面外用药物作用后,倒置显微镜观察其生长变化、流式细胞仪检测其细胞周期、二甲基噻唑二苯基四唑溴盐(MTT)法检测其细胞增殖等的变化。结果细胞形态学观察、二甲基噻唑二苯基四唑溴盐(MTT)吸光度OD值(optical density)、细胞增殖指数[PI=(S+G2/M)/(G0/G1+S+G2/M)]值的结果与对照组比较结果较一致:庆大霉素组、碱性成纤维细胞生长因子(basic fibrob last growth factor,bFGF)组、丁胺卡那霉素0.021μg/m l组Hacat细胞变化均不明显(P>0.05);其余各种剂量抗生素组及表皮生长因子(ep iderm al growth factor,EGF)组抑制Hacat细胞增值,数目较少,形态不规则,透明度低,细胞内多有颗粒样物质及空泡。OD值、PI值均明显降低(P<0.05,P<0.01)。其中氯霉素组、丁胺卡那霉素组OD值存在组内差异,低浓度组对细胞增殖抑制作用较中、高浓度低(P<0.05)。重组人生长激素(recomb inant hum an growth hormone,rhGH)组促进Hacat细胞增值,OD值、PI值与对照组比较均增高(P<0.05),且存在组内差异,高浓度组对细胞增殖促进作用较中、低浓度低(P<0.05)。结论①不同创面外用药物对Hacat细胞生物学特性影响各异。②氯霉素组、丁胺卡那霉素组、rhGH组对Hacat细胞生物学特性的影响存在组内差异。

Objective To observe the effect of different kinds of external application agents in different concentrations on the biological characteristics of human adult skin keratinocyte propagated under low Ca and elevated temperature （Hacat）. Methods Hacat cells were passaged in vitro. The growth rate, cell cycle and the proliferation of the cultured Hacat cells were determined by inverted microscope, flow cytometry and MTT respectively, after the cells were cocultured with the external agents for burn wounds including some antibiotics, bFGF, EGF and rhGH. Results The morphology, the optical density （OD） by MTT, cell cycle by flow cytometry expressed as proliferation index ——PI = （S ＋ G2/M ）/（ G0/G1 ＋ S ＋ G2/M ） of Hacat cells treated with gentamicin, bFGF and 0. 021μg/ml amikacin were of no significant difference with that of control cells. But OD value or PI of the Hacat cells decreased significantly （ P 〈 0. 05, P 〈 0. 01 ） in all other antibiotic （ in all concentrations） and EGF groups, with granules and vacuoles within Hacat cells of irregular shape and clear contour. There was intra-group difference of OD value in chloromycetin and amikacin groups. The proliferation was more significant, the OD value and PI were much higher in the Hacat cells in rhGH groups than that in control group （ P 〈 0. 05 ） with obvious intra-group difference, rhGH of moderate and low concentrations exerted more powerful promoting effect on the proliferation of Hacat cells than rhGH of high concentration （ P 〈 0. 05 ）. Conclusion Different kinds of external application agents for burn wounds exerted different effects on the biological characteristics of Hacat cells. There was significant intra-group difference in the effects of some agents on the biological characteristics of Hacat cells, such as chloromycetin, amikacin and rhGH.