CAJ | 학술논문

目的探讨微波与小胶质细胞促炎症反应之间的关系,揭示小胶质细胞在微波致中枢神经损伤中的作用.方法离体培养的N9小胶质细胞接受90 mW/cm2微波辐照,在辐照后0、1、3、6、12、24 h采用细胞流式计数的方法观察CD11b的表达情况,用酶联免疫吸附(EUSA)方法检测N9细胞培养上清液中TNF-α的浓度,采用反转录-聚合酶链反应(RT-PCR)检测N9小胶质细胞一氧化氮合酶(iNOS)mRNA表达,采用硝酸还原酶法检测培养上清液中NO含量.结果微波辐照后3 h阳性表达CD11b的小胶质细胞明显增加,与对照组比较,差异有统计学意义(P<0.01),一直持续到辐照后24 h,并在辐照后6 h达到高峰.TNF-α含量在辐射后1 h明显升高[(657.8±25.9)pg/ml],并一直持续到24 h,在辐照后3 h达到峰值[(762.1±61.5)pg/ml],与对照组[(258.9±81.7)pg/ml]比较,差异有统计学意义(P<0.01.NO的含量在辐照后1 h开始升高(4.48±0.59) μmol/L].与对照组[(2.65±0.14) μmol/L]的差异有统计学意义(P<0.05),并随时间的延长逐渐增加.iNOS mRNA表达在辐照后1 h开始明显升高,6 h升高约2倍,此后表达有所下降,但24 h内均高于对照组,差异均有统计学意义(P<0.05或P<0.01).结论微波辐照可明显诱导小胶质细胞活化,活化的小胶质细胞可通过分泌大量TNF-α,释放NO等产生促炎症反应.
목적 탐토미파여소효질세포촉염증반응지간적관계,게시소효질세포재미파치중추신경손상중적작용.방법 리체배양적N9소효질세포접수90 mW/cm2미파복조,재복조후0、1、3、6、12、24 h채용세포류식계수적방법관찰CD11b적표체정황,용매련면역흡부(EUSA)방법검측N9세포배양상청액중TNF-α적농도,채용반전록-취합매련반응(RT-PCR)검측N9소효질세포일양화담합매(iNOS)mRNA표체,채용초산환원매법검측배양상청액중NO함량.결과 미파복조후3 h양성표체CD11b적소효질세포명현증가,여대조조비교,차이유통계학의의(P<0.01),일직지속도복조후24 h,병재복조후6 h체도고봉.TNF-α함량재복사후1 h명현승고[(657.8±25.9)pg/ml],병일직지속도24 h,재복조후3 h체도봉치[(762.1±61.5)pg/ml],여대조조[(258.9±81.7)pg/ml]비교,차이유통계학의의(P<0.01.NO적함량재복조후1 h개시승고(4.48±0.59) μmol/L].여대조조[(2.65±0.14) μmol/L]적차이유통계학의의(P<0.05),병수시간적연장축점증가.iNOS mRNA표체재복조후1 h개시명현승고,6 h승고약2배,차후표체유소하강,단24 h내균고우대조조,차이균유통계학의의(P<0.05혹P<0.01).결론 미파복조가명현유도소효질세포활화,활화적소효질세포가통과분비대량TNF-α,석방NO등산생촉염증반응.
Objective To explore the relationship between microglia] proinflammatory and electro-magnetic radiation and unveil the role of microglia in microwave radiation induced central nervous system in-jury. Methods N9 microglia cells cultured in vitro were exposed to microwave at 90 mW/cm2. Cell flow cy-tometry was used to observe the expression of CD1 lb at different time points after exposure; ELISA was used to detect the concentration of TNF-α in N9 cell culture supernatant; RT-PCR analysis confirmed iNOS mRNA ex-pression in N9 microglia cells; and Nitrate Reductase Method was used to test NO amount in culture super-natant. Results The CD11b positive microglial cells increased significantly at 3 h after microwave exposure (P<0.05), continued to increase until 24 h and peaked at 6 h after exposure. The amount of TNF-α rose dra-matically from 1 h to 24 h after exposure (P<0.01) and peaked at 3 h [(762.1±61.5) pg/ml] after exposure (P< 0.01). The level of NO started to increase at 1 h [(4.48±0.59) μmol/L] and lasted for 24 h after exposure. The expression of iNOS mRNA increased significantly at 1 h(P<0.05), and tripled the original expression at 6 h af-ter exposure, hereafter, it decreased slightly, but all were higher than the control group within 24 h after exposure. Conclusion Microwave radiation could induce the activation of microglia cells. The activated microglia cells could induce microglial proinflammatory by producing large amounts of TNF-α, NO, etc.