钙离子和氯化钴促进成纤维细胞色素上皮衍生因子的表达

Calcium and cobalt chloride enhance the expression of pigment epithelium-derived factor in fibroblasts

目的探讨色素上皮衍生因子(PEDF)在成纤维细胞中的表达以及钙离子和氯化钴对其表达的影响。方法 19例包皮标本根据年龄分为2组:13例40岁以下者为I组,6例40岁以上者为II组。以I型胶原酶分离真皮成纤维细胞,37oC5%CO2培养。以1.0mmol/L氯化钙和100μmol/L的氯化钴作用于培养的成纤维细胞。以免疫荧光法检测PEDF蛋白质的表达,以逆转录-聚合酶链反应(RT-PCR)检测PEDF mRNA的表达情况。结果 PEDF表达于正常成纤维细胞,而且在组I和组II间mRNA相对表达量无差异性(分别为,P>0.05)。1.0mmol/L氯化钙和100μmol/L氯化钴显著增加PEDF mRNA相对表达量,分别为2.3倍和1.4倍,二者差异具有显著性(P<0.05)。结论真皮成纤维细胞表达PEDF,而且受钙离子和氯化钴的调节。

Objective To investigate the expression of PEDF and its regulation by calcium and cobalt chloride（CoCl2） in dermal fibroblasts.Methods Ninteen normal human foreskin specimens were grouped into I or II according to their ages below（n=13） or above 40（n=6）.Dermal fibroblasts were separated by collagenase type I and cultured at 37oC in 5% CO2.Calcium at 1.0 mM and CoCl2 at 100μM were incubated with fibroblasts for 24h.The mRNA and protein level of PEDF was determined by RT-PCR and immunofluorescence respectively.Results The expression of PEDF in normal fibroblasts was identified by RT-PCR and immunofluorescence,and no difference of PEDF mRNA was detected between groups I and II（P0.05）.Calcium at 1.0 mM and CoCl2 at 100μM significantly enhanced the expression of PEDF in cultured fibroblasts about 2.3 and 1.4 fold,respectively（P0.05）.Conclusion PEDF is expressed in dermal fibroblasts and is regulated by calcium and CoCl2.