【摘要】 目的:研究防御素作用于慢性牙周炎致病微生物分子机制。方法:将HBD-1,-2和-3以及HBD-1,-2和-3 siRNAs转染细胞后RT-PCR检测IL-1β,IL-6,IL-8,IL-10和TNF-α的表达水平。利用过表达HBD-1,-2和-3的293细胞培养上清液与Pg、Tf、Td、Fn和Aa共同培养分别观察过表达HBD-1,-2和-3的上清液的抑菌效果并进行细菌克隆计数,计算克隆抑制率。结果:转染HBD-1,-2和-3病毒载体后,HBD-1可以显著抑制IL-6和IL-1β的表达; HBD-2可以显著降低TNF-α和IL-1β的表达; HBD-3可以显著降低TNF-α,IL-6和IL-8的表达,而HBD-2和HBD-3显著增加了IL-10的表达。转染siRNAs后,其抑制能力减弱。HBD-1,-2和-3可以不同程度抑制Pg、Tf、Td、Fn和Aa的增殖。结论:HBD-1,-2和-3抑制并降低L-1β,IL-6,IL-8和TNF-α等促炎因子表达,对牙周炎主要致病微生物增殖具有一定抑制作用,提示利用基因工程技术将HBD-1,-2和-3用于牙周炎治疗与预防可行。更多还原

【Abstract】 Objective: To explore how to use human beta-defensin( HBD) in the prevention and treatment of periodontitis,this experiment aimed to study HBD on the molecular mechanism of chronic periodontitis of pathogenic microorganism. Methods: The relative mRNA levels of IL-1β,IL-6,IL-8,IL-10 and TNF-α were detected by RT-PCR after keratinocytes transfected with HBD-1,-2 and-3 or HBD-1,-2 and-3 siRNAs. The supernatant fluids from 293 cells overexpressing HBD-1,-2 and-3 were co-cultured with Pg,Tf,Td,Fn and Aa. To observe the bacteriostatic effect of HBD-1,-2 and-3 overexpressed supernatants,the bacterial colonies and clone inhibition rate were counted and calculated. Results: The expression of IL-6 and IL-1β could be significantly inhibited by HBD-1. The expression of TNF-α and IL-1β could be reduced by HBD-2. TNF-α and IL-6 and IL-8 express levels could be suppressed by HBD-3. But IL-10 expression significantly increased by HBD-2 and HBD-3. After transfected siRNAs,this effect was significantly attenuated. HBD-1,-2 and-3 could inhibit the proliferation of Pg,Tf,Td,Fn and Aa. Conclusion: HBD-1,-2 and-3 decreased the expression of some proinflammatory cytokines and inhibited the proliferation of the main pathogenic microorganisms in periodontitis at different levels,suggesting that HBD-1,-2 and-3 can be used in the treatment and prevention of periodontitis by genetic engineering.更多还原