CAJ | 학술논문

目的探讨伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)脂多糖诱导兔不同部位的单核巨噬细胞细胞因子表达的差异,以期为研究牙周炎与全身疾病间可能的免疫机制提供理论基础.方法分离5只新西兰兔外周血单核巨噬细胞(peripheral mononuclear cells,Mo)、肺单核巨噬细胞(alveolar macrophages,AM)、腹腔单核巨噬细胞(peritoneal macrophages,PM)及肝脏单核巨噬细胞(Kupffer cells,KC),对每个部位的细胞分别用1 mg/L大肠杆菌-脂多糖(大肠杆菌-脂多糖组)、Aa-脂多糖刺激(Aa-脂多糖组),以不加任何刺激的细胞为空白对照组,每组均为6个样本.24 h后采用实时PCR及酶联免疫吸附测定法分别检测肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白介素(interleukin,IL)6、IL-1β、IL-8 mRNA及蛋白表达.结果新西兰兔各部位单核巨噬细胞大肠杆菌-脂多糖组及Aa-脂多糖组4种细胞因子mRNA及蛋白表达均显著高于空白对照组(P＜0.05),其中AM的Aa-脂多糖组TNF-α、1L-6、IL-1β、IL-8 mRNA[分别为(0.4719±0.0171)、(2.7895±0.0669)、(5.1527±0.1190)、(3.6785±0.1836)]及蛋白[分别为(82.2±5.4)、(40.2±2.0)、(50308.3±445.0)、(35 305.3±1480.9) ng/L]的相对表达均最强；Aa-脂多糖组4种细胞因子mRAN及蛋白表达均显著强于大肠杆菌-脂多糖组(P＜0.05)；不同部位的单核巨噬细胞对Aa-脂多糖的反应存在部位差异性(P＜0.05),总体上从强至弱依次为AM、Mo、KC、PM.结论 Aa-脂多糖对兔不同部位的单核巨噬细胞各细胞因子的表达存在影响,且这种影响具有部位差异性.
목적 탐토반방선방선간균(Actinobacillus actinomycetemcomitans,Aa)지다당유도토불동부위적단핵거서세포세포인자표체적차이,이기위연구아주염여전신질병간가능적면역궤제제공이론기출.방법 분리5지신서란토외주혈단핵거서세포(peripheral mononuclear cells,Mo)、폐단핵거서세포(alveolar macrophages,AM)、복강단핵거서세포(peritoneal macrophages,PM)급간장단핵거서세포(Kupffer cells,KC),대매개부위적세포분별용1 mg/L대장간균-지다당(대장간균-지다당조)、Aa-지다당자격(Aa-지다당조),이불가임하자격적세포위공백대조조,매조균위6개양본.24 h후채용실시PCR급매련면역흡부측정법분별검측종류배사인자α(tumor necrosis factor-α,TNF-α)、백개소(interleukin,IL)6、IL-1β、IL-8 mRNA급단백표체.결과 신서란토각부위단핵거서세포대장간균-지다당조급Aa-지다당조4충세포인자mRNA급단백표체균현저고우공백대조조(P＜0.05),기중AM적Aa-지다당조TNF-α、1L-6、IL-1β、IL-8 mRNA[분별위(0.4719±0.0171)、(2.7895±0.0669)、(5.1527±0.1190)、(3.6785±0.1836)]급단백[분별위(82.2±5.4)、(40.2±2.0)、(50308.3±445.0)、(35 305.3±1480.9) ng/L]적상대표체균최강；Aa-지다당조4충세포인자mRAN급단백표체균현저강우대장간균-지다당조(P＜0.05)；불동부위적단핵거서세포대Aa-지다당적반응존재부위차이성(P＜0.05),총체상종강지약의차위AM、Mo、KC、PM.결론 Aa-지다당대토불동부위적단핵거서세포각세포인자적표체존재영향,차저충영향구유부위차이성.
Objective To investigate the expression of cytokines induced by Actinobacillus actinomycetemcomitans lipopolysaccharide(Aa-LPS) in monocytes/macrophages from different organs of rabbits.Methods The peripheral mononuclear cells(Mo),alveolar macrophages (AM),peritoneal macrophages(PM) and Kupffer cells (KC) from five New Zealand rabbits were isolated respectively.Then the cells from different organs were stimulated with Escherichia coli (Ec)-LPS or Aa-LPS at the dose of 1 mg/L.Ater culture for 24 hours,the expression of tumor necrosis factor-α (TNF-α),interleukin (IL) 6,IL-1β,IL-8 mRNA and protein were determined by real-time PCR and enzyme-linked immunosorbent assay respectively.Results The monocytes/macrophages challenged by Ec-LPS or Aa-LPS expressed more cytokines both in mRNA and protein levels compared with the controls (P ＜ 0.05).Among them,AM displayed the highest respond when encount with Aa-LPS,with the TNF-α,IL-6,IL-1β,1L-8 mRNA relative levels were(0.4719 ±0.0171),(2.7895 ± 0.0669),(5.1527 ±0.1190),(3.6785 ±0.1836) and the proteins concentrations were (82.2 ±5.4),(40.2 ±2.0),(50308.3 ±445.0),(35305.3 ± 1480.9) ng/L respectively.And the inducibility of Aa-LPS was stronger than that of Ec-LPS (P ＜ 0.05).Meanwhile the cells from different organs showed discrepant response when exposed to Aa-LPS (P ＜ 0.05).The results showed their abilities to secrete cytokines were in the sequence of AM ＞ Mo ＞ KC ＞ PM.Conclusions Aa-LPS enfluenced the expression of cytokines in monocytes/macrophages from different organs of rabbits.