CAJ | 학술논문

背景：体外实验表明，淫羊藿苷可抑制脂多糖诱导的急性肺炎，其抗炎作用在磨损颗粒存在的条件下是否依然有效？目的：通过体内外实验相结合的方法探讨淫羊藿苷对磨损颗粒诱导炎症反应的调控作用。方法：①体内实验：将80只雄性C57BL/6小鼠随机分为4组，钛组和钛+淫羊藿苷组采用钛诱导小鼠颅骨无菌炎症模型，对照组、淫羊藿苷组也进行相同手术过程，但不植入钛，淫羊藿苷组、钛+淫羊藿苷组建模当天灌胃给予淫羊藿苷200 mg/(kg·d)，对照组、钛组灌胃给予等量的安慰剂，2周后酶联免疫吸附实验、定量RT-PCR检测肿瘤坏死因子α，白细胞介素1β蛋白或mRNA的表达量。②体外实验：将小鼠单核/巨噬细胞RAW264.7分别与核因子кB受体配体、核因子кB受体配体+淫羊藿苷、核因子кB受体配体+钛颗粒及核因子кB受体配体+钛颗粒+淫羊藿苷共培养72 h，ELISA检测培养基上清中肿瘤坏死因子α，白细胞介素1β水平，RT-PCR分析肿瘤坏死因子α，白细胞介素1β基因mRNA表达。结果与结论：①体内实验：在钛颗粒存在的条件下，口服淫羊藿苷可明显减少颗粒诱导的炎症细胞浸润，使炎性增厚的骨膜变薄，抑制颅骨标本中肿瘤坏死因子α，白细胞介素1β的表达。②体外实验：经钛颗粒刺激后，细胞培养基中肿瘤坏死因子α，白细胞介素1β质量浓度显著增加，细胞中相应mRNA表达量上调，而经淫羊藿苷干预后这两种炎性因子在蛋白和基因水平的表达量显著下调。结果表明淫羊藿苷在体内外均可显著抑制钛颗粒诱导的炎症反应。
배경：체외실험표명，음양곽감가억제지다당유도적급성폐염，기항염작용재마손과립존재적조건하시부의연유효？목적：통과체내외실험상결합적방법탐토음양곽감대마손과립유도염증반응적조공작용。방법：①체내실험：장80지웅성C57BL/6소서수궤분위4조，태조화태+음양곽감조채용태유도소서로골무균염증모형，대조조、음양곽감조야진행상동수술과정，단불식입태，음양곽감조、태+음양곽감조건모당천관위급여음양곽감200 mg/(kg·d)，대조조、태조관위급여등량적안위제，2주후매련면역흡부실험、정량RT-PCR검측종류배사인자α，백세포개소1β단백혹mRNA적표체량。②체외실험：장소서단핵/거서세포RAW264.7분별여핵인자кB수체배체、핵인자кB수체배체+음양곽감、핵인자кB수체배체+태과립급핵인자кB수체배체+태과립+음양곽감공배양72 h，ELISA검측배양기상청중종류배사인자α，백세포개소1β수평，RT-PCR분석종류배사인자α，백세포개소1β기인mRNA표체。결과여결론：①체내실험：재태과립존재적조건하，구복음양곽감가명현감소과립유도적염증세포침윤，사염성증후적골막변박，억제로골표본중종류배사인자α，백세포개소1β적표체。②체외실험：경태과립자격후，세포배양기중종류배사인자α，백세포개소1β질량농도현저증가，세포중상응mRNA표체량상조，이경음양곽감간예후저량충염성인자재단백화기인수평적표체량현저하조。결과표명음양곽감재체내외균가현저억제태과립유도적염증반응。
BACKGROUND:Studiesin vitro have suggested that icarin can attenuate lipopolysaccharide (LPS)-induced acute pneumonia. Is the anti-inflammatory effect of icarin stil valid in the presence of wear particles? OBJECTIVE:With studiesin vivo andin vitro, to investigate the regulatory effect of icarrin on titanium particle-induced inflammatory reaction. METHODS:(1) Studiesin vivo: Eighty male C57BL/6 mice aged 6-8 weeks were randomly divided into four groups: control group, icarin group, titanium particle group, and titanium particle+icarin group. Mice in the titanium particle group and titanium particle+icarin group received surgical procedure, and sterile and endotoxin-free titanium particles were implanted on the calvaria surfaces to induce inflammatory reaction. Mice in the control group and icarin group received the same surgery, but no wear particles were implanted. Then icarin was given oraly to mice in the titanium particle group and titanium particle+ icarin group with a dose of 200 mg/kg per day for 2 weeks from the day of modeling. Mice in the control group and icarin group were given oraly the same dose of placebo. Two weeks later, tumor necrosis factor-α and interleukin-1β at protein and mRNA levels were respectively detected with enzyme-linked immunohistochemical (ELISA) and quantitative real time reverse transcription PCR (qRT-PCR) analysis. (2) Studiesin vitro: Mouse monocyte/macrophage RAW264.7 cels were cultured with different conditioned media: control group, nuclear factor receptor ligand кB (RANKL); icarin group, RANKL+icarin; titanium particle group, RANKL+titanium particles; titanium particle+icarrin group, RANKL+icarin+titanium particles. Titanium particles stimulated RAW264.7 cels were co-cultured with RANKL and icarin for 72 hours. Tumor necrosis factor-α and interleukin-1β at protein and mRNA levels in the supernatant were detected with ELISA analysis and qRT-PCR, respectively. RESULTS AND CONCLUSION: (1) Resultsin vivo: icarin treatment obviously decreased titanium particle-induced inflammatory cellinfiltration and made the thickness of periosteum thinner, down-regulated tumor necrosis factor-α and interleukin-1β expressions at protein and mRNA levels. (2) Results in vitro: when RAW264.7 cels were stimulated with titanium particles for 72 hours, tumor necrosis factor-α and interleukin-1β expressions at protein and mRNA levels in culture media increased obviously; when icarin was administrated, tumor necrosis factor-α and interleukin-1βexpressions at protein and mRNA levels down-regulated significantly. These results suggest icarin can obviously suppress titanium particle-induced inflammatory reactionin vivo andin vitro.