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

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。

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

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 epithehal cells (hESGc).Methods The first generation of hESGc were cultured in keratinocyte serum free medium (KSFM) and treated with various concentrations (2,20,40μg/L) of HGF for different durations.Then,cell scratch test was performed to detect cell migration,a double staining flow cytometry assay using annexin VFITC/propidium iodide to detect cell apoptosis.and Western blot to measure the expression of p-Akt.Results HGF of 2μg/L had no effect on the migration of hESGc,while that of 20 μg/L and 40μg/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 40μg/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 40μ/L, respectively;there was a significant difference between untreated cells and 40μg/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.