中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (6): 411-414.

• 论著 • 上一篇    下一篇

肝细胞生长因子对体外培养的人外泌汗腺腺上皮细胞迁移、凋亡及p-Akt表达的影响

雷霞1,刘渤2,伍津津3,鲁元刚1,朱堂友4,杨亚东1   

  1. 1. 第三军医大学大坪医院皮肤科
    2.
    3. 重庆第三军医大学大坪医院皮肤科
    4. 第三军医大学大坪医院
  • 收稿日期:2009-09-17 修回日期:2010-01-31 出版日期:2010-06-15 发布日期:2010-06-08
  • 通讯作者: 雷霞 E-mail:leixia1979@sina.com
  • 基金资助:

    国家自然科学基金;第三军医大学校级课题

Effect of hepatocyte growth factor on the migration and apoptosis of, as well as p-Akt expression in cultured human eccrine sweat gland epithelial cells

  • Received:2009-09-17 Revised:2010-01-31 Online:2010-06-15 Published:2010-06-08
  • Contact: xia lei E-mail:leixia1979@sina.com

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摘要：

目的 研究肝细胞生长因子（HGF）对人外泌汗腺上皮细胞（hESGc）的迁移、凋亡及p-Akt表达的影响。方法 在无血清培养基中分别加入不同浓度HGF（2、20、40 μg/L）培养hESGc，采用细胞损伤愈合模型，培养20 h后计数进入刮痕区的细胞，观察HGF 对hESGc移行的作用。采用Annexin V-FITC及PI双染（膜联蛋白-异硫氰酸荧光素及碘化丙啶双染）后经流式细胞仪检测细胞凋亡，观察HGF对hESGc凋亡的影响，并用Western印迹法检测细胞中p-Akt的变化。结果 2 μg/L HGF无促细胞移行作用，20 μg/L和40 μg/L HGF组进入划痕区的细胞分别为（23.0 ± 6.3）个和（56.7 ± 7.9）个，与对照组（17.3 ± 5.5）个相比，差异均有统计学意义（t值分别为2.653、15.858，P值 < 0.05或0.01），两种浓度HGF促移行能力分别为33.2%和228.2％，差异有统计学意义（P < 0.01）。与对照组凋亡率14.76%相比，2 μg/L HGF和20 μg/L HGF组细胞凋亡率无明显变化，其凋亡率分别为14.16%和13.50%，而40 μg/L HGF组细胞凋亡率为8.87%，显著下降（t = 7.852，P < 0.01）。Western印迹法检测显示加入HGF后Akt被磷酸化，p-Akt升高。结论 HGF能促进hESGc移行，抑制hESGc凋亡，并刺激p-Akt表达升高。

关键词: 肝细胞生长因子, 外泌汗腺, 上皮细胞, 细胞移行, 细胞凋亡

Abstract:

Objective To investigate the effect of hepatocyte growth factor （HGF） on migration and apoptosis of, as well as phosphorylated-Akt （p-Akt） expression in cultured human eccrine sweat gland epithelial cells （hESGc）. Methods The first generation of hESGc were cultured in keratinocyte serum free medium （KSFM） and treated with various concentrations （2, 20, 40 μg/L） of HGF for different durations. Then, cell scratch test was performed to detect cell migration, a double staining flow cytometry assay using annexin V-FITC/propidium iodide to detect cell apoptosis, and Western blot to measure the expression of p-Akt. Results HGF of 2 μg/L had no effect on the migration of hESGc, while that of 20 μg/L and 40 μg/L could promote the migration of hESGc by 33.2% and 228.2%, respectively. The average number of cells migrating into the scrach zone was significantly lower in untreated cell group than that in 20 and 40 μg/L HGF-treated cell group （17.3 ± 5.5 vs 23.0 ± 6.3 and 56.7 ± 7.9, t = 2.653, 15.858, P < 0.05, 0.01, respectively）. The apoptosis rate was 14.76% in untreated cells, 14.16%, 13.5% and 8.87% in cells treated with HGF of 2, 20 and 40 μg/L, respectively; there was a significant difference between untreated cells and 40 μg/L HGF-treated cells （t = 7.852, P < 0.01）. HGF could activate the phosphorylation of Akt protein and increase the expression of p-Akt. Conclusion HGF could promote the migration of，inhibit the apoptosis of, and stimulate the p-Akt expression in, hESGc.

Key words: Hepatocyte growth factor, Eccrine sweat gland, Epithelial cells, cell migration, cell apoptosis