过表达PTEN对LPS诱导的成纤维细胞周期及胶原分泌的影响

Influence of excessive PTEN expression to fibroblast cycle and collagen secretion induced by LPS

目的探索过表达第10号染色体丢失的张力蛋白同源的磷酸酶基因(PTEN)对大肠杆菌细菌脂多糖(LPS)诱导的成纤维细胞(FB)周期及胶原分泌的影响。方法体外培养增生性瘢痕患者正常皮肤成纤维细胞,原代培养接近80%融合时消化传代,培养至第3代,采用0.5μg/mL的LPS刺激第3代正常皮肤成纤维细胞,将携带有PTEN基因的缺陷型腺病毒转染LPS刺激后的第3代正常皮肤成纤维细胞。采用流式细胞仪检测细胞周期,ELISA法检测细胞胶原分泌。结果过表达PTEN基因能够抑制LPS处理后成纤维细胞周期G2M期的增加。LPS刺激能够升高皮肤成纤维细胞中胶原的分泌,过表达PTEN能够抑制LPS诱导的细胞胶原的分泌。结论 LPS可增加成纤维细胞G2M期的数量,增加胶原分泌,过表达PTEN可阻断这种作用。

Objective To explore the influence of excessive PTEN expression to fibroblast cycle and collagen secretion induced by LPS.Methods Normal skin fibroblast in the patient with hyperplastic scar were cultured in vitro.When the primary culture was close to 80%fusion,the digestive passage was performed,cultured to the third generation.LPS(0.5μg/mL)was adopted to stimulate the third generation of normal skin fibroblasts.Defective adenovirus carrying PTEN gene was transfected to the third passage fibroblasts after LPS stimulation.Flow cytometer was adopted to detect the cell cycle.ELISA method was adopted to detect the secreted collagen amont.Results Excessive PTEN expression could inhibit the increase of G2 Mcell cycle induced by LPS.LPS stimulation could increase the secretion of collagen in skin fibroblasts,yet excessive PTEN expression could inhibit the secretion of collagen induced by LPS.Conclusion LPS could increase the amont of fibroblasts on G2 Mcell cycle and secretion of collagen,yet excessive PTEN expression can inhibit the effect.