体外培养人牙周膜成纤维细胞雌激素受体表达的实验研究

The expression of estrogen receptors in the human periodontal ligament fibroblasts in vitro

目的观察雌激素受体在体外培养的人牙周膜成纤维细胞中的表达,为探讨雌激素对牙周组织的作用提供依据。方法用免疫细胞化学法对原代细胞爬片雌激素受体(estrogenreceptors,ER)-α、β行 SABC 染色;用反转录聚合酶链反应法检测 ER-α、ER-β mRNA 在人牙周膜成纤维细胞中的表达;用 Western blot 法检测 ER-α、ER-β蛋白的表达。结果 ER-α和 ER-β在人牙周膜成纤维细胞中均有表达,ER-β表达强于 ER-α。分别计算 ER-α、ER-β对应蛋白条带灰度值与内参β-肌动蛋白条带灰度值的比值,得到 A 组细胞 ER-α、ER-β相对灰度值的均值为(0.80±0.08)和(1.05±0.11),B 组细胞则为(0.95±0.09)和(1.13±0.10),ER-α和 ER-β相比差异有统计学意义(P<0.05),说明 ER-β的条带较 ER-α对应的蛋白条带浓,即 ER-β蛋白的相对丰度大于 ER-α。但对于同一受体亚型,无论是 ER-α或 ER-β,A、B 组间蛋白条带灰度值差异均无统计学意义(P>0.05),说明雌激素非处理组和经雌激素处理组在 ER 蛋白表达水平上无明显差异。结论雌激素可能通过其受体 ER-α、ER-β对人牙周膜成纤维细胞的生物学性状产生影响。

Objective To explore the expression of estrogen receptors in the human periodontal ligament fibroblasts in vitro. Methods Human periodontal ligament fibroblasts were cultured in vitro. The estrogen receptors （ER）-α, ER-β were detected with immunocytochemistry staining; The mRNA of ER-α,ER-β were measured by reverse transcription polymerase chain reaction; The expression of ER-α, ER-β protein were detected by Western blot. Results The mRNA and protein expression of two subtype of ER were observed on human periodontal ligament fibroblasts（HPLF）. The intensity of the ER-β bands were stronger than those of ER-α, indicating that ER-β expression in HPLF was higher. Conclusions ER may play an important role in the function of human periodontal ligament fibroblasts.