四川大学学报（医学版）
四川大學學報（醫學版）
사천대학학보（의학판）
JOURNAL OF SICHUAN UNIVERSITY(MEDICAL SCIENCE EDITION)
2009年
6期
1011-1014
,共4页
Aβ_(1-42)%U251细胞%一氧化氮%RANTES%NF-κB%STAT1
Aβ_(1-42)%U251細胞%一氧化氮%RANTES%NF-κB%STAT1
Aβ_(1-42)%U251세포%일양화담%RANTES%NF-κB%STAT1
β-amyloid peptide 1-42%U251 cell%Nitric oxide%RANTES%NF-κB%STAT1
目的 探讨β淀粉样蛋白1-42(Aβ_(1-42))体外诱导人神经胶质瘤细胞(U251)细胞炎性反应的分子学机制. 方法 U251细胞经常规去血清培养,采用终浓度为0.2～4.0 μmol/L Aβ_(1-42)处理U251细胞24 h,以四唑盐比色(MTT)法测定细胞存活率;2 μmol/L Aβ_(1-42)处理U251细胞12、24、36、48 h后,采用硝酸还原酶法检测一氧化氮(NO)含量;采用双抗体夹心ELISA法测定细胞RANTES蛋白表达水平;免疫细胞化学染色法检测NF-κB和STAT1的表达.结果 随着Aβ_(1-42)剂量的增加,细胞的存活率降低(P<0.05);用2 μmol/L Aβ_(1-42)处理U251细胞12、24、36、48 h,结果显示NO的生成于24 h达高峰,此后逐渐下降;同浓度Aβ_(1-42)处理U251细胞24 h后,RANTES的表达增加4倍(P<0.01);2 μmol/L Aβ_(1-42)刺激U251细胞24 h后细胞内NF-κB P65和STAT1从胞浆移至胞核且表达明显. 结论 Aβ_(1-42)降低体外培养的U251细胞存活率,诱导NO和RANTES的释放,提示Aβ_(1-42)诱导的U251细胞趋化因子RANTES产生可能与NF-κB和STAT1的活化有关.
目的 探討β澱粉樣蛋白1-42(Aβ_(1-42))體外誘導人神經膠質瘤細胞(U251)細胞炎性反應的分子學機製. 方法 U251細胞經常規去血清培養,採用終濃度為0.2～4.0 μmol/L Aβ_(1-42)處理U251細胞24 h,以四唑鹽比色(MTT)法測定細胞存活率;2 μmol/L Aβ_(1-42)處理U251細胞12、24、36、48 h後,採用硝痠還原酶法檢測一氧化氮(NO)含量;採用雙抗體夾心ELISA法測定細胞RANTES蛋白錶達水平;免疫細胞化學染色法檢測NF-κB和STAT1的錶達.結果 隨著Aβ_(1-42)劑量的增加,細胞的存活率降低(P<0.05);用2 μmol/L Aβ_(1-42)處理U251細胞12、24、36、48 h,結果顯示NO的生成于24 h達高峰,此後逐漸下降;同濃度Aβ_(1-42)處理U251細胞24 h後,RANTES的錶達增加4倍(P<0.01);2 μmol/L Aβ_(1-42)刺激U251細胞24 h後細胞內NF-κB P65和STAT1從胞漿移至胞覈且錶達明顯. 結論 Aβ_(1-42)降低體外培養的U251細胞存活率,誘導NO和RANTES的釋放,提示Aβ_(1-42)誘導的U251細胞趨化因子RANTES產生可能與NF-κB和STAT1的活化有關.
목적 탐토β정분양단백1-42(Aβ_(1-42))체외유도인신경효질류세포(U251)세포염성반응적분자학궤제. 방법 U251세포경상규거혈청배양,채용종농도위0.2～4.0 μmol/L Aβ_(1-42)처리U251세포24 h,이사서염비색(MTT)법측정세포존활솔;2 μmol/L Aβ_(1-42)처리U251세포12、24、36、48 h후,채용초산환원매법검측일양화담(NO)함량;채용쌍항체협심ELISA법측정세포RANTES단백표체수평;면역세포화학염색법검측NF-κB화STAT1적표체.결과 수착Aβ_(1-42)제량적증가,세포적존활솔강저(P<0.05);용2 μmol/L Aβ_(1-42)처리U251세포12、24、36、48 h,결과현시NO적생성우24 h체고봉,차후축점하강;동농도Aβ_(1-42)처리U251세포24 h후,RANTES적표체증가4배(P<0.01);2 μmol/L Aβ_(1-42)자격U251세포24 h후세포내NF-κB P65화STAT1종포장이지포핵차표체명현. 결론 Aβ_(1-42)강저체외배양적U251세포존활솔,유도NO화RANTES적석방,제시Aβ_(1-42)유도적U251세포추화인자RANTES산생가능여NF-κB화STAT1적활화유관.
Objective To study the molecular mechanisms of β-amyloid 1-42 (A(β_(1-42)) induced the inflammatory response in U251 cells. Methods U251 cells were treated with different concentration of Aβ_(1-42)(0. 2-4. 0 μmol/L) for 24 h, and the cell survival rate was evaluated by MTT. After treated with 2. 0μmol/L Aβ_(1-42) for 12,24,36,48 hours, the nitric oxide content of U251 cells were detected with nitrate reductase method. The RANTES expression was tested by dual-antibody sandwich ELISA method. The expression of NF-ΚB and STAT_1 were detected by the immunocytochemical staining. Results Aβ_(1-42) decreased the survival rate of U251 cells in dose-dependent manner. The NO generated in 2μmol/L A.β_(1-42) treated U251 cells reached its peak at 24 h and gradually decreased thereafter. The expression of RANTES increased 4-fold (P<0. 01). The NF-ΚB P65, the STAT_1 moved from the cytoplasm to the nucleus and expressed significantly were also observed in the 2μmol/L Aβ_(1-42) treated U251 cells. Conclusion Aβ1-42 decreases the survival rate of U251 cells and induce the releasing of NO and RANTES. This indicates that Aβ_(1-42) induced chemokine RANTES in U251 cell may be close related to the activation of NF-κB and STAT1.
