中华皮肤科杂志 ›› 2007, Vol. 40 ›› Issue (2): 95-97.

• 论著 • 上一篇    下一篇

HaCaT细胞对红色毛癣菌生长的抑制作用

陈辉, 刘维达, 吕桂霞, 余晓东, 沈永年   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042
  • 收稿日期:2006-03-06 发布日期:2007-02-15

Inhibitory effect of HaCaT cells on the growth of Trichophyton rubrum in vitro

CHEN Hui, LIU Wei-da, LU Gui-xia, SHE Xiao-dong, SHEN Yong-nian   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2006-03-06 Published:2007-02-15

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摘要： 目的 探讨角质形成细胞对红色毛癣菌生长的抑制作用.方法 将HaCaT细胞采用含10%小牛血清的DMEM培养液,5%CO2环境下,37℃孵育至80%融合时,弃去培养液,加入用无血清DMEM配制的菌丝段悬液共孵育(浓度0.1×105/mL～1.0×105/mL),并以菌丝段悬液作为阴性对照.分别于4 h、8 h、16 h和24 h制备共孵育后的菌丝段悬液,进行台盼蓝染色计数和菌落计数,对真菌生长状况进行评价并计算真菌生长抑制率.结果 在细胞和菌丝段共孵育4 h、8 h、16 h和24 h,台盼蓝染色法测得的菌生长抑制百分数分别为46.8%±8.0%、64.0%±5.5%、76.3%±7.7%和39.5%±4.9%;菌落计数法测得菌生长抑制百分数分别为39.7%±4.9%、58.0%±8.9%、68.3%±7.4%和39.9%±5.5%.结论 HaCaT细胞对体外培养的红色毛癣菌有一定的抑制作用.

关键词: 角蛋白细胞, 红色毛癣菌, 细胞, 培养的

Abstract: Objective To investigate the influence of keratinocytes on the growth of Trichophyton rubrum in vitro.Methods The hyphae suspension of T.rubrum was prepared in Dulbecco's Modified Eagle's Medium (DMEM) without bovine serum at working concentration of 0.1×105/mL to 1.0×105/mL,and preserved at 4℃.HaCaT cells,an immortal human keratinocyte cell line,were grown in DMEM supplemented with 10% calf serum at 37℃ in humidified air with 5% CO2 until 80% confluency.After removel of the cell culture supematant,the hyphae suspension was added to coculture with HaCaT cells,with the suspension alone as a negative control.The hyphae suspension was harvested after 4,8,16,and 24 h of conculture,and fungal viability was determined by trypan blue staining and enumeration of the colony-forming units.Finally,the growth inhibitory rates of Trichophyton rubrum at each time point were calculated.Results With trypan blue staining,the growth inhibitory rates of Trichophyton rubrum at 4,8,16,and 24 h were 46.8±8.0%,64.0±5.5%,76.3±7.7%,and 39.5±4.9%,respectively.Enumeration of the colony-forming units revealed that the inhibitory rates were 39.7±4.9%,58.0±8.9%,68.3±7.4%,and 39.9±5.5% at 4,8,16,and 24 h,respectively.Conclusion HaCaT cells can inhibit the growth of Trichophyton rubrum in vitro.

Key words: Keratinocytes, Trichophyton rubrum, Cells, cultured