摘要
目的探讨正常皮肤和增生性瘢痕组织成纤维细胞第10号染色体丢失的张力蛋白同源的磷酸酶基因(phosphatase and tensin homolog deleted on chromosome 10,PTEN)表达差异及大肠杆菌细菌脂多糖(lipopolysaccharides,LPS)刺激对PTEN表达的影响。方法体外培养增生性瘢痕患者正常皮肤(对照组)和瘢痕组织(瘢痕组)成纤维细胞,采用QPCR检测2组成纤维细胞PTEN mRNA表达水平,采用Western blot法检测2组成纤维细胞PTEN蛋白表达水平。正常成纤维细胞原代培养约80%融合时消化传代,培养至第3代,采用0、0.005、0.01、0.1、0.5mg/L LPS进行刺激,并对刺激后细胞传代培养至第8代,采用Western blot法检测不同浓度LPS刺激下成纤维细胞PTEN蛋白表达水平。结果瘢痕组成纤维细胞PTEN mRNA和蛋白(0.29±0.03、0.38±0.04)表达水平明显低于对照组(0.99±0.02、2.56±0.02)(P<0.05);0.5mg/L LPS刺激正常皮肤成纤维细胞PTEN蛋白表达水平(0.29±0.02)低于0、0.005、0.01、0.1mg/L LPS刺激(1.01±0.15、0.71±0.02、0.59±0.01、0.32±0.01)(P<0.05),随LPS浓度增高,PTEN表达量逐渐下降。结论瘢痕组织成纤维细胞PTEN表达低于正常皮肤成纤维细胞,LPS可降低正常成纤维细胞PTEN表达。
Objective To explore the PTEN expression difference between normal skin and hyperplastic scar and the influence of lipopolysaceharides （LPS） from Escherichia coli on PTEN expression. Methods Fibroblasts of normal skin and hyperplastic scar were cultured in vitro. QPCR and Western blot methods were adopted to detect the expressions of PTEN mRNA and protein. Normal skin fibroblasts were cultured to the third passage and to the eighth passage after exposed to 0, 0. 005, 0.01, 0. 1 and 0.5 mg/L LPS. The PTEN expression levels under different LPS concentrations were detected by Western blot method. Results The levels of PTEN mRNA and protein （0. 29±0.03, 0. 38±0. 04） in fibroblasts of hyperplastic scar were significantly lower than those in fibroblasts of normal skin （0.99 ± 0.02, 2.56 ± 0.02） （P〈0.05）. PTEN protein expression was significantly lower under the stimulation of 0.5 mg/L LPS （0.29 ± 0.02） than that under 0, 0. 005, 0.01 and 0.1 mg/L LPS （1.01±0.15, 0.71±0.02, 0. 59±0. 01, 0.32±0.01） （P〈 0.05）. PTEN expression decreased gradually with the increase of LPS concentration. Conclusion PTEN expression in fibroblasts of hyperplastic scar is lower than that of normal skin. LPS could reduce the PTEN expression in normal fibroblasts.
出处
《中华实用诊断与治疗杂志》
2016年第11期1116-1118,共3页
Journal of Chinese Practical Diagnosis and Therapy
基金
海军总医院创新培育基金(2014)
