白细胞介素-1β对遗传性牙龈纤维瘤和正常牙龈上皮细胞β-防御素的影响

Effect of interlukine-1β on the expression of HBD-1,2,3 in epithelial cell of normal gingival and hereditary gingival fibromatosis

目的:比较白细胞介素1β(IL-1β)刺激体外培养的正常牙龈、遗传性牙龈纤维瘤(HGF)上皮细胞β-防御素(HBD)表达的差异,探讨该差异与遗传性牙龈纤维瘤发病机制的可能相关性。方法:体外培养3例遗传性牙龈纤维瘤病人和6例正常人牙龈上皮细胞,经0,0.01,0.1,1,10,100 ng/mL的IL-1β分别刺激12、24、36、48 h,提取细胞总RNA,逆转录后,采用HBD-1、2、3特异性引物经PCR扩增,以β-肌动蛋白为内参,计算HBD-1、2、3与内参扩增产物相对值,对HBD-1、2、3进行半定量分析,采用SPSS软件单向方差统计分析法分析结果。结果:HBD-1 mRNA在正常牙龈和遗传性牙龈纤维瘤上皮细胞中呈固有表达,不受IL-1β刺激的影响。IL-1β可上调两种牙龈上皮细胞HBD-2、3的表达,在浓度为0.1～10 ng/mL时,作用时间大于12 h,受刺激组与未受刺激组差异有显著性(P<0.01)。相同浓度IL-1β作用不同时间时,正常牙龈上皮细胞中HBD-2、3的表达水平显著高于遗传性牙龈纤维瘤上皮细胞中表达水平(P<0.05,P<0.01)。且随作用时间延长差异更加显著。结论:遗传性牙龈纤维瘤病变组织和正常牙龈组织中β-防御素的表达有差异,这种差异可能与遗传性牙龈纤维瘤上皮细胞的分化及发病机制相关。

AIM： To analyze the expression of HBD - 1,2,3mRNA in normal gingival （NG） and HGF epithelial cells, and investigate the effect of IL - 1β on the expression of HBD - 1,2,3 gene by NG and HGF epithelial cell. METHDOS ： Epithelial cells from NF and HGF were cultured and induced by IL - 1β with of different concentrations of 0,0.01,0.1,1,10,100ng/mL with 0,12,24,36 or 48 hours. The effect of IL- 1β on HBD- 1,2,3 mRNA were examined by RT - PCR and β - actin was used as an internal invariant control. RESULTS ： HBD - 1 mRNA was inherent expression on epithelial cell from NF and HGF. IL - 1β increased the expression of HBD -2,3 mRNA with dose - depended at 0. 1 - 10ng/ml and over 12 hours. There were significant different HBD -2,3 gene expression between non - stimulated and stimulated group （P 〈 0.01 ）. The level of HBD -2,3 on NF epithelial cell were significantly higher than that of HGF under IL - 1 β effect（P 〈0.05 ,P 〈0.01 ）. CONCLUSION： The different patterns of HBD - 1,2,3 expression between NF and HGF epithelial cells may be related with the differentiation and pathogenesis of HGF.