中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
35期
2826-2829
,共4页
张英%雍慧媛%史小婷%周俊%马千%连庆泉%曹红%李军
張英%雍慧媛%史小婷%週俊%馬韆%連慶泉%曹紅%李軍
장영%옹혜원%사소정%주준%마천%련경천%조홍%리군
阿尔茨海默病%姜黄素%β-淀粉样蛋白%PC12细胞
阿爾茨海默病%薑黃素%β-澱粉樣蛋白%PC12細胞
아이자해묵병%강황소%β-정분양단백%PC12세포
Alzheimer's disease%Curcumin%Aβ25-35%PC12 cell
目的 探讨姜黄素(Cur)对AD细胞活力及HMGB1表达的影响.方法 对数期生长的PC12细胞分为5组:空白组(A组)不做处理,Aβ 25-35模型组(B组)加入20μmol/L的Aβ25-35,Aβ25-35+Cur治疗组(C组)加入20 μmol/L的Aβ25-35和1μmol/L的Cur,Aβ25-35+ rHMG1损伤组(D组)加入20 μmol/L的Aβ25-35和500 ng/ml的HMGB1,Aβ25-35+溶剂对照组(E组)加入20 μmol/L的Aβ25-35和1μl/ml的DMSO,孵育24h后行细胞形态学观察、免疫荧光定性和免疫蛋白印迹(Western blot)法检测HMGB1蛋白的表达情况.结果 与A组相比,B、D、E组细胞活力明显下降(0.76±0.06、0.63±0.02、0.75±0.03比1.22 ±0.06,P＜0.05)、胞内HMGB1表达明显升高(1.19±0.14、1.12±0.16、1.16±0.09比0.85±0.04,P＜0.05)；与B组相比,C组细胞活力上升33％、HMGB1表达下降31％ (P ＜0.05).B组较A组存在大量的HMGB1核外释放,而C组HMGB1的核外释放较B组减少.结论 姜黄素可减轻Aβ25-35引起的PC12细胞毒性,其机制与下调HMGB1的表达、抑制HMGB1的核外释放有关.
目的 探討薑黃素(Cur)對AD細胞活力及HMGB1錶達的影響.方法 對數期生長的PC12細胞分為5組:空白組(A組)不做處理,Aβ 25-35模型組(B組)加入20μmol/L的Aβ25-35,Aβ25-35+Cur治療組(C組)加入20 μmol/L的Aβ25-35和1μmol/L的Cur,Aβ25-35+ rHMG1損傷組(D組)加入20 μmol/L的Aβ25-35和500 ng/ml的HMGB1,Aβ25-35+溶劑對照組(E組)加入20 μmol/L的Aβ25-35和1μl/ml的DMSO,孵育24h後行細胞形態學觀察、免疫熒光定性和免疫蛋白印跡(Western blot)法檢測HMGB1蛋白的錶達情況.結果 與A組相比,B、D、E組細胞活力明顯下降(0.76±0.06、0.63±0.02、0.75±0.03比1.22 ±0.06,P＜0.05)、胞內HMGB1錶達明顯升高(1.19±0.14、1.12±0.16、1.16±0.09比0.85±0.04,P＜0.05)；與B組相比,C組細胞活力上升33％、HMGB1錶達下降31％ (P ＜0.05).B組較A組存在大量的HMGB1覈外釋放,而C組HMGB1的覈外釋放較B組減少.結論 薑黃素可減輕Aβ25-35引起的PC12細胞毒性,其機製與下調HMGB1的錶達、抑製HMGB1的覈外釋放有關.
목적 탐토강황소(Cur)대AD세포활력급HMGB1표체적영향.방법 대수기생장적PC12세포분위5조:공백조(A조)불주처리,Aβ 25-35모형조(B조)가입20μmol/L적Aβ25-35,Aβ25-35+Cur치료조(C조)가입20 μmol/L적Aβ25-35화1μmol/L적Cur,Aβ25-35+ rHMG1손상조(D조)가입20 μmol/L적Aβ25-35화500 ng/ml적HMGB1,Aβ25-35+용제대조조(E조)가입20 μmol/L적Aβ25-35화1μl/ml적DMSO,부육24h후행세포형태학관찰、면역형광정성화면역단백인적(Western blot)법검측HMGB1단백적표체정황.결과 여A조상비,B、D、E조세포활력명현하강(0.76±0.06、0.63±0.02、0.75±0.03비1.22 ±0.06,P＜0.05)、포내HMGB1표체명현승고(1.19±0.14、1.12±0.16、1.16±0.09비0.85±0.04,P＜0.05)；여B조상비,C조세포활력상승33％、HMGB1표체하강31％ (P ＜0.05).B조교A조존재대량적HMGB1핵외석방,이C조HMGB1적핵외석방교B조감소.결론 강황소가감경Aβ25-35인기적PC12세포독성,기궤제여하조HMGB1적표체、억제HMGB1적핵외석방유관.
Objective To explore the effects of curcumin on the expression of high mobility group box1 (HMGB1),cell viability and morphology in a cellular model of Alzheimer's disease (AD).Methods Cultured PC12 cells in logarithmic growth phase were divided into 5 groups:normal cell group (A,nontreatment),model control group (B,20 μmol/L Aβ25-35),curcumin treatment group (C,20 μmol/LAβ25-35 + 1 μmol/L Cur),Aβ25-35 + rHMG1 (D,20 μ mol/L Aβ25-35 + 500 ng/ml HMGB1) and solvent control group (E,20 μ mol/L Aβ25-35 + 1 μl/ml DMSO).Cell viability was examined by methyl thiazolyl tetrazolium (MTT).And the cellular expression and distribution of HMGB1 were detected by immunofluorescence and Western blot 24 hours later.Results Compared with group A,the levels of cell viability in groups B,D and E significantly declined (0.76 ± 0.06,0.63 ± 0.02,0.75 ± 0.03 vs 1.22 ±0.06,P＜0.05) while the expression of HMGB1 increased (1.19 ±0.14,1.12 ±0.16,1.16 ±0.09 vs 0.85 ± 0.04,P ＜ 0.05).Compared with group B,cell viability in group C significantly increased by 33％ (1.01 ± 0.05,P ＜ 0.05) while the expression of HMGB1 declined by 31％ (0.78 ± 0.03,P ＜ 0.05).A larger amount of extracellular HMGB1 was released in group B compared with group A.And the extracellular release of HMGB1 declined less in group C versus group B.Conclusion Curcumin may reduce Aβ25-35-induced cytotoxicity through a down-regulated expression of HMGB1 and an inhibition of extracellular release of HMGB1 in PC12 cell.
