中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
9期
697-700
,共4页
神经胶质瘤%细胞系%白细胞介素-1β%环氧化酶-2%磷脂酶A2,胞质型
神經膠質瘤%細胞繫%白細胞介素-1β%環氧化酶-2%燐脂酶A2,胞質型
신경효질류%세포계%백세포개소-1β%배양화매-2%린지매A2,포질형
Glioma%Cells line%Inlerleukin-1 beta%Cyclooxygenase%Phospholipases A2,cytosolic
目的 探讨白细胞介素(IL)-1β调节人胶质细胞瘤内环氧合酶(COX)-2系统表达的分子机制.方法 以2.5 μg/L IL-1 β刺激人胶质瘤细胞H4细胞,在刺激培养的12、24、48和72 h收集细胞及上清液.以不同浓度的p38和p42/44 MAPK信号抑制剂SB203580和PD98059分别预处理H4细胞1h后进行IL-1β刺激培养24 h,收集细胞及上清液.Northern印迹杂交法及免疫印迹法检测H4细胞中胞质型磷脂酶A(cPLA)2和COX-2蛋白的表达情况;ELISA法检测细胞培养上清液中前列腺素(PGE)2的含量.结果 2.5 μg/L IL-1β以时间依赖性方式诱导H4细胞中cPLA2和COX-2的表达,在刺激培养24 h时影响最为明显(P＜0.05),之后有下降趋势.同时,IL-1β还能诱导PGE2的产生,刺激培养72 h时PGE2产量增加250倍;p38 MAPK抑制剂SB203580能以剂量依赖性(0.1、1和10 μmol/L)有效抑制IL-1β诱导的cPLA2和COX-2表达(P＜0.05),而p42/44 MAPK抑制剂PD98059则对二者表达影响不明显(P＞0.05).但是,SB203580和PD98059均能显著降低IL-1β诱导PGE2的产生(P＜0.05).结论 IL-1β能明显诱导COX-2和cPLA2的表达,并通过p38和p42/44MAPK信号通路介导PGE2的产生,因此在翻译后水平上影响PGE2的表达.
目的 探討白細胞介素(IL)-1β調節人膠質細胞瘤內環氧閤酶(COX)-2繫統錶達的分子機製.方法 以2.5 μg/L IL-1 β刺激人膠質瘤細胞H4細胞,在刺激培養的12、24、48和72 h收集細胞及上清液.以不同濃度的p38和p42/44 MAPK信號抑製劑SB203580和PD98059分彆預處理H4細胞1h後進行IL-1β刺激培養24 h,收集細胞及上清液.Northern印跡雜交法及免疫印跡法檢測H4細胞中胞質型燐脂酶A(cPLA)2和COX-2蛋白的錶達情況;ELISA法檢測細胞培養上清液中前列腺素(PGE)2的含量.結果 2.5 μg/L IL-1β以時間依賴性方式誘導H4細胞中cPLA2和COX-2的錶達,在刺激培養24 h時影響最為明顯(P＜0.05),之後有下降趨勢.同時,IL-1β還能誘導PGE2的產生,刺激培養72 h時PGE2產量增加250倍;p38 MAPK抑製劑SB203580能以劑量依賴性(0.1、1和10 μmol/L)有效抑製IL-1β誘導的cPLA2和COX-2錶達(P＜0.05),而p42/44 MAPK抑製劑PD98059則對二者錶達影響不明顯(P＞0.05).但是,SB203580和PD98059均能顯著降低IL-1β誘導PGE2的產生(P＜0.05).結論 IL-1β能明顯誘導COX-2和cPLA2的錶達,併通過p38和p42/44MAPK信號通路介導PGE2的產生,因此在翻譯後水平上影響PGE2的錶達.
목적 탐토백세포개소(IL)-1β조절인효질세포류내배양합매(COX)-2계통표체적분자궤제.방법 이2.5 μg/L IL-1 β자격인효질류세포H4세포,재자격배양적12、24、48화72 h수집세포급상청액.이불동농도적p38화p42/44 MAPK신호억제제SB203580화PD98059분별예처리H4세포1h후진행IL-1β자격배양24 h,수집세포급상청액.Northern인적잡교법급면역인적법검측H4세포중포질형린지매A(cPLA)2화COX-2단백적표체정황;ELISA법검측세포배양상청액중전렬선소(PGE)2적함량.결과 2.5 μg/L IL-1β이시간의뢰성방식유도H4세포중cPLA2화COX-2적표체,재자격배양24 h시영향최위명현(P＜0.05),지후유하강추세.동시,IL-1β환능유도PGE2적산생,자격배양72 h시PGE2산량증가250배;p38 MAPK억제제SB203580능이제량의뢰성(0.1、1화10 μmol/L)유효억제IL-1β유도적cPLA2화COX-2표체(P＜0.05),이p42/44 MAPK억제제PD98059칙대이자표체영향불명현(P＞0.05).단시,SB203580화PD98059균능현저강저IL-1β유도PGE2적산생(P＜0.05).결론 IL-1β능명현유도COX-2화cPLA2적표체,병통과p38화p42/44MAPK신호통로개도PGE2적산생,인차재번역후수평상영향PGE2적표체.
Objective To explore the molecular mechanisms by which interleukin-1 β (IL-1β) regulates the expression of cytosolic phospholipase A2 (cPLA2) and cyclooxygenase-2 (COX-2) in human neuroglioma cell.Methods H4 neuroglioma cells were treated with IL-1 β (2.5 μg/L) for different timepoints up to 72 h.For MAPK study,cells were incubated for 1 h with MAPK inhibitors,SB203580 and PD98059 and subsequently stimulated with IL-1β (1 μg/L) for 24 h.Northern and Western blot were used to determine the protein expressions of cPLA2 and COX-2 respectively.And the content of PGE2 in supernatant was determined by enzyme-linked immunosorbent assay (ELISA).Results A dose of 2.5 μg/L IL-1 β induced the protein expressions of cPLA2 and COX-2 and a subsequent release of PGE2 in a time-dependent manner.And the expressions of cPLA2 and COX-2 peaked at 24 h after stimulation (P ＜ 0.05).The expression of PGE2 increased 250 folds after a 72 h culture.Both SB203580 and PD98059 inhibitors reduced IL-1 β-induced PGE2 production while SB203580 alone reduced the expressions of both cPLA2 and COX-2.Conclusion IL-1β induces the expressions of cPLA2 and COX-2 and affects COX-2 at the post-translational level by modulating PGE2 production through the signal transduction pathways of p38 and p42/44 MAPKs.
