LPS及IL-1β对组织工程化口腔黏膜中HBD-2和HBD-3表达的影响  

The effects of LPS and IL-1βon the expression of HBD-2 and HBD-3 in tissue engineered oral mucosa

作　　者：汪洋[1] 宣静[1] 王海燕[1] 黄艺[1] 项美娟 WANG Yang;XUAN Jing;WANG Hai-yan;HUANG Yi;XIANG Mei-juan(Department of Stomatology,Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine.Shanghai 200071,China)

机构地区：[1]上海中医药大学附属上海市中医医院口腔科,上海200071

出　　处：《中国口腔颌面外科杂志》2024年第6期536-540,共5页China Journal of Oral and Maxillofacial Surgery

摘　　要：目的:研究在大肠杆菌脂多糖(lipopolysaccharide,LPS)和重组人白细胞介素1(interleukin-1,IL-1β)刺激下,组织工程化口腔黏膜替代品中的人β防御素2(Human-beta-defensin-2,HBD-2)和人β防御素3(Human-beta-defensin-3,HBD-3)的表达,评估组织工程化口腔黏膜治疗感染性口腔疾病的可能性。方法:采用实时荧光定量反转录PCR(RT-qPCR)和免疫印迹试验(Western blot)评估在LPS和IL-1β刺激下,组织工程化口腔黏膜替代品中HBD-2和HBD-3基因及蛋白的表达情况。采用SPSS 26.0软件包对数据进行统计学分析。结果:LPS和IL-1β均可刺激组织工程化口腔黏膜中HBD-2和HBD-3基因及蛋白的表达,培养24 h后呈现出剂量依赖性增加的趋势(P<0.05)。在100 ng/mL的LPS刺激下,HBD-2基因表达增加约2.7倍,而HBD-3基因表达增加约5倍;在20 ng/mL的IL-1β刺激下,HBD-2基因表达增加约1.6倍,而HBD-3基因表达增加约4.8倍。Western blot结果显示,LPS和IL-1β均可显著诱导HBD-2和HBD-3蛋白的分泌。结论:组织工程化口腔黏膜替代品在一定程度上具备抵御入侵微生物的防御功能,应用组织工程化口腔黏膜可为感染性口腔疾病提供新的治疗思路。PURPOSE:To investigate the expression of human beta-defensin 2(HBD-2)and human beta-defensin 3(HBD-3)in tissue engineered oral mucosa equivalents under the stimulation of Escherichia coli lipopolysaccharide(LPS)and recombinant human interleukin-1(IL-1β),and to evaluate the possibility of using tissue engineered oral mucosa in the treatment of infectious oral diseases.METHODS:Real-time fluorescent quantitative reverse transcription PCR(RT-qPCR)and Western blot were used to evaluate the mRNA and protein expression of HBD-2 and HBD-3 in tissue engineered oral mucosa substitutes under the stimulation of LPS and IL-1β.SPSS 26.0 software package was used for data analysis.RESULTS:LPS and IL-1βstimulated the mRNA and protein expression of HBD-2 and HBD-3 in tissue-engineered oral mucosa,and showed a dose-dependent increase after 24 hours of culture(P<0.05).At 100 ng/mL,LPS induced HBD-2 mRNA by about 2.7-fold and HBD-3 mRNA by about 5-fold,respectively.The maximal production of HBD-2 mRNA was about 1.6-fold and HBD-3 about 4.8-fold by 20 ng/mL IL-1β.As to the results of western blot analysis,both LPS and IL-1βinduced HBD-2 protein secretion at the maximal concentration in this study.HBD-3 protein was hardly observed without any treatment.After incubation with LPS and IL-1β,HBD-3 protein was significantly up-regulated(P<0.05).CONCLUSIONS:Tissue engineered oral mucosa equivalents are armed with defense activity against invading microorganisms in a sense,which may provide a therapy substitute for infectious oral diseases in the future.

关 键 词：组织工程化口腔黏膜 HBD-2 HBD-3 免疫防御 

分 类 号：R739.8[医药卫生—肿瘤]