目的：观察不同浓度细菌脂多糖(BLPS)作用下原代培养的人牙龈上皮细胞(HGECs)β防御素-4(BD-4)的表达。方法体外原代培养HGECs,经免疫细胞化学鉴定,加入10、20、40、80、160μg/mL的BLPS,采用免疫组织化学和Western blot-ting检测BD-4蛋白在HGECs中的定位和表达变化。结果 HGECs组织块法原代细胞生长状态好,第3代细胞进行鉴定,Ck (AE1/AE3)(+)/Vimentin(-)。 HGECs免疫细胞化学染色结果显示,HGECs胞浆可见棕黄色颗粒状着色,阳性染色弥漫分布于胞浆,随着BLPS浓度的增加表达更为强烈。 Western Blotting结果显示,BD-4表达水平随BLPS浓度增加而增高。 BD-4蛋白在BLPS浓度为160μg/mL表达的值最高,组间比较差异有统计学意义(P<0.01)。结论随着BLPS对牙龈上皮炎症刺激加重,BD-4表达增强,提示了BD-4在牙周炎症进程中的作用。%Objective To study the expression ofβ-defensin 4 in primary human gingival epithelial cells ( HGECs) after bacterial li-popolysaccharide ( BLPS) inducement. Methods Cultured in vitro, primary HGECs were identified by immunocytochemistry. BLPS with the concentration of 10, 20, 40, 80 or 160 μg/mL was added to HGECs. Protein expression of BD-4 in HGECs was measured by immunocytochemistry and western blotting. Results HGECs grew well. The third generation cells were identified Ck ( AE1/AE3 ) (+)/Vimentin(-) . By immunocytochemistry, the positively stained granular brown was distributed in cytoplasm, and the expression was more intense with the increase of BLPS concentration. By western blotting, the expression of BD-4 increased with higher concentra-tion of BLPS. The top value of BD-4 protein was presented in 160μg/mL BLPS. There were significant differences between the HGECs groups of BLPS inducement (P<0. 01). Conclusion With the exacerbation of inflammation in HGECs induced by BLPS, the expres-sion of BD-4 increases, suggesting the function of BD-4 in periodontal disease.
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