中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2010年
6期
682-686
,共5页
罗鸣%伍钢%范丽%张瑞光%任精华%董继华%董晓荣
囉鳴%伍鋼%範麗%張瑞光%任精華%董繼華%董曉榮
라명%오강%범려%장서광%임정화%동계화%동효영
Corilagin%小胶质细胞%NF-κB%放射性脑损伤
Corilagin%小膠質細胞%NF-κB%放射性腦損傷
Corilagin%소효질세포%NF-κB%방사성뇌손상
Corilagin%Microglia BV-2%NF-κB%Radiation encephlopathy
目的 研究抗炎药物Corilagin抑制放射引起小胶质细胞(BV-2)炎性反应的作用及其防护的分子机制.方法 细胞抑制实验(MTT)检测Corilagin对BV-2细胞的抑制率;Corilagin预处理小胶质细胞,12 h后,以0和32 Gy 2个放射剂量照射BV-2细胞,Real-time PCR检测小胶质细胞照射后不同时间点炎性反应因子IL-1β、TNF-α表达水平;硝酸还原酶法检测细胞上清液中一氧化氮(NO)的含量;Western blot检测各组细胞核转录因子NF-κB p65蛋白表达;激光共聚焦显微镜观察各组细胞标志物Iba-1的表达、NF-κB p65核转位及Nemo的表达.结果 1～10μg/ml浓度范围内Corilagin对BV-2细胞增殖几乎无影响;照射后小胶质细胞表面标志物Iba-1表达明显,提示小胶质细胞激活,且炎性反应因子NO、TNF-α、IL-1β表达上调,而Corilagin(5μg/ml)可以显著抑制上述炎性反应因子的表达(tIL-1β=6.341,tTNF-α=3.411,tNO=3.134,P＜0.05);Corilagin可抑制NF-κB活化蛋白Nemo,并显著抑制NF-κB p65的转位.结论 Corilagin可能通过NF-κB信号转导途径抑制照射后小胶质细胞的活化,从而下调炎性反应因子表达,保护神经元.
目的 研究抗炎藥物Corilagin抑製放射引起小膠質細胞(BV-2)炎性反應的作用及其防護的分子機製.方法 細胞抑製實驗(MTT)檢測Corilagin對BV-2細胞的抑製率;Corilagin預處理小膠質細胞,12 h後,以0和32 Gy 2箇放射劑量照射BV-2細胞,Real-time PCR檢測小膠質細胞照射後不同時間點炎性反應因子IL-1β、TNF-α錶達水平;硝痠還原酶法檢測細胞上清液中一氧化氮(NO)的含量;Western blot檢測各組細胞覈轉錄因子NF-κB p65蛋白錶達;激光共聚焦顯微鏡觀察各組細胞標誌物Iba-1的錶達、NF-κB p65覈轉位及Nemo的錶達.結果 1～10μg/ml濃度範圍內Corilagin對BV-2細胞增殖幾乎無影響;照射後小膠質細胞錶麵標誌物Iba-1錶達明顯,提示小膠質細胞激活,且炎性反應因子NO、TNF-α、IL-1β錶達上調,而Corilagin(5μg/ml)可以顯著抑製上述炎性反應因子的錶達(tIL-1β=6.341,tTNF-α=3.411,tNO=3.134,P＜0.05);Corilagin可抑製NF-κB活化蛋白Nemo,併顯著抑製NF-κB p65的轉位.結論 Corilagin可能通過NF-κB信號轉導途徑抑製照射後小膠質細胞的活化,從而下調炎性反應因子錶達,保護神經元.
목적 연구항염약물Corilagin억제방사인기소효질세포(BV-2)염성반응적작용급기방호적분자궤제.방법 세포억제실험(MTT)검측Corilagin대BV-2세포적억제솔;Corilagin예처리소효질세포,12 h후,이0화32 Gy 2개방사제량조사BV-2세포,Real-time PCR검측소효질세포조사후불동시간점염성반응인자IL-1β、TNF-α표체수평;초산환원매법검측세포상청액중일양화담(NO)적함량;Western blot검측각조세포핵전록인자NF-κB p65단백표체;격광공취초현미경관찰각조세포표지물Iba-1적표체、NF-κB p65핵전위급Nemo적표체.결과 1～10μg/ml농도범위내Corilagin대BV-2세포증식궤호무영향;조사후소효질세포표면표지물Iba-1표체명현,제시소효질세포격활,차염성반응인자NO、TNF-α、IL-1β표체상조,이Corilagin(5μg/ml)가이현저억제상술염성반응인자적표체(tIL-1β=6.341,tTNF-α=3.411,tNO=3.134,P＜0.05);Corilagin가억제NF-κB활화단백Nemo,병현저억제NF-κB p65적전위.결론 Corilagin가능통과NF-κB신호전도도경억제조사후소효질세포적활화,종이하조염성반응인자표체,보호신경원.
Objective To explore the inhibitory effects of Corilagin on the production of proinflammatory cytokines in microglia induced by radiation. Methods The cytotoxicity of Corilagin was measured by MTT assay. Microglia BV-2 cells were irradiated 0 or 32 Gy after pretreated with Corilagin for 12 hours. Realtime-PCR was used to detect the mRNA levels of inflammatory cytokines, such as IL-1β,TNF-α on several time-points. The content of nitric oxide (NO) was determined with nitrate reductase method. The translocation of NF-κB was measured by Western blot and immunocytochemical stain.Confocal microscopy was used to observe the expression of Iba-1 and Nemo. Results No cytotoxicity was detected on BV-2 cells with 1-10 μg/ml Corilagin. Iba-1 expression in microglia cells was activated by irradiation, the expression levels of inflammatory cytokines, such as IL-1β, TNF-α and NO were also elevated. Whereas, the production of IL-1 β, TNF-α in activated microglia cells was significantly inhibited with 5 μg/mL corilagin ( tIL-1β = 6. 341, tTNF-α = 3.41 1, tNO = 3. 134, P ＜ 0. 05 ). Corilagin significantly inhibited the expression of Nemo and the translocation of NF-κB p65. Conclusion Corilagin could inhibit the activation of irradiated microglia cells and down-regulate the expression of inflammatory cytokines, via inhibition of the NF-κB signaling pathway.
