中华行为医学与脑科学杂志
中華行為醫學與腦科學雜誌
중화행위의학여뇌과학잡지
CHINESE JOURNAL OF BEHAVIORAL MEDICINE AND BRAIN SCIENCE
2012年
2期
123-125
,共3页
王凤斌%丁振禹%孔佑华%王亚南%李宁
王鳳斌%丁振禹%孔祐華%王亞南%李寧
왕봉빈%정진우%공우화%왕아남%리저
三羟基异黄酮%APP695%PC12细胞%细胞周期%细胞凋亡
三羥基異黃酮%APP695%PC12細胞%細胞週期%細胞凋亡
삼간기이황동%APP695%PC12세포%세포주기%세포조망
Genistein%APP695%PC12 cells%Cell cycle%Apoptosis
目的 以APP695MT基因转染PC12细胞为细胞模型,研究三羟基异黄酮(Genistein,GST)对模型细胞周期和凋亡的影响.方法 用pIRES2-EGFP空载体和pIRES2-EGFP/APP695 MT表达载体转染正常的PC12细胞,将细胞分为空载对照组、APP695转染组、GST干预组.应用流式细胞仪测定检测细胞周期和细胞凋亡率,激光共聚焦显微镜观察细胞凋亡形态学变化.结果 APP695转染组与空载对照组比较,PC12细胞在G0和G1期明显增多,而进入S期的细胞减少,细胞增殖指数明显降低[(55.6±0.57)％,P＜0.01],细胞凋亡率明显升高[(77.10±12.53)％,P＜0.01]；细胞死亡发出的红色荧光明显增多,胞体肿胀,细胞器崩解,细胞核固缩或碎裂.GST( 15 μmol/L)干预组与APP695转染组比较,PC12细胞在Go和G1期明显减少,而进入S期的细胞增多,细胞增殖指数明显增加[ (61.57±0.47)％,P＜0.01],细胞凋亡率降低[(46.00±8.43)％,P＜0.01]；细胞死亡发出的红色荧光明显减少,绿色荧光显著增强,细胞形态较APP695转染组好转.结论 GST能改善APP695MT基因转染引起的细胞周期阻滞,促进细胞向S期的过渡,降低PC12细胞凋亡率,对转染细胞有一定的保护作用.
目的 以APP695MT基因轉染PC12細胞為細胞模型,研究三羥基異黃酮(Genistein,GST)對模型細胞週期和凋亡的影響.方法 用pIRES2-EGFP空載體和pIRES2-EGFP/APP695 MT錶達載體轉染正常的PC12細胞,將細胞分為空載對照組、APP695轉染組、GST榦預組.應用流式細胞儀測定檢測細胞週期和細胞凋亡率,激光共聚焦顯微鏡觀察細胞凋亡形態學變化.結果 APP695轉染組與空載對照組比較,PC12細胞在G0和G1期明顯增多,而進入S期的細胞減少,細胞增殖指數明顯降低[(55.6±0.57)％,P＜0.01],細胞凋亡率明顯升高[(77.10±12.53)％,P＜0.01]；細胞死亡髮齣的紅色熒光明顯增多,胞體腫脹,細胞器崩解,細胞覈固縮或碎裂.GST( 15 μmol/L)榦預組與APP695轉染組比較,PC12細胞在Go和G1期明顯減少,而進入S期的細胞增多,細胞增殖指數明顯增加[ (61.57±0.47)％,P＜0.01],細胞凋亡率降低[(46.00±8.43)％,P＜0.01]；細胞死亡髮齣的紅色熒光明顯減少,綠色熒光顯著增彊,細胞形態較APP695轉染組好轉.結論 GST能改善APP695MT基因轉染引起的細胞週期阻滯,促進細胞嚮S期的過渡,降低PC12細胞凋亡率,對轉染細胞有一定的保護作用.
목적 이APP695MT기인전염PC12세포위세포모형,연구삼간기이황동(Genistein,GST)대모형세포주기화조망적영향.방법 용pIRES2-EGFP공재체화pIRES2-EGFP/APP695 MT표체재체전염정상적PC12세포,장세포분위공재대조조、APP695전염조、GST간예조.응용류식세포의측정검측세포주기화세포조망솔,격광공취초현미경관찰세포조망형태학변화.결과 APP695전염조여공재대조조비교,PC12세포재G0화G1기명현증다,이진입S기적세포감소,세포증식지수명현강저[(55.6±0.57)％,P＜0.01],세포조망솔명현승고[(77.10±12.53)％,P＜0.01]；세포사망발출적홍색형광명현증다,포체종창,세포기붕해,세포핵고축혹쇄렬.GST( 15 μmol/L)간예조여APP695전염조비교,PC12세포재Go화G1기명현감소,이진입S기적세포증다,세포증식지수명현증가[ (61.57±0.47)％,P＜0.01],세포조망솔강저[(46.00±8.43)％,P＜0.01]；세포사망발출적홍색형광명현감소,록색형광현저증강,세포형태교APP695전염조호전.결론 GST능개선APP695MT기인전염인기적세포주기조체,촉진세포향S기적과도,강저PC12세포조망솔,대전염세포유일정적보호작용.
Objective To investigate the effect of Genistein (GST) on cell cycle and apoptosis in PC12 cells transfected App695MT gene.Methods PC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid,and then were divided into control vectortransfected group,APP695 transfected group and GST treatment group.Flow cytometry was applied to detect cell cycle and apoptosis,laser confocal microscope was used to observe morphological changes of cell apoptosis.Results Compared with control vectortransfected group,PC12 cells in APP695 transfected group increased significantly in G0 and G1 phase,and less into S phase,cell proliferation index was decreased significantly( (55.6 ±0.57)％,P＜0.0l ),apoptosis rate was increased significantly( (77.10 ± 12.53)％,P＜0.01 ).Emitted red fluorescence increased significantly when cell death,cell body swelling,organelle disintegration,nuclear condensation or fragmentation.Compared with APP695 transfected group,PC12 cells in GST( 15μ mol/L) treatment group,decreased significantly in G0 and G1 phase,and more into S phase,cell proliferation index was increased significantly ( ( 61.57 ± 0.47 ) ％,P ＜ 0.01 ),apoptosis rate was decreased significantly ( (46.00 ± 8.43 ) ％,P ＜ 0.01 ).Cell death was significantly reduced red fluorescence,emitted green fluorescence was significantly enhanced,compared with APP695 transfected group cell morphology improved.Conclusion GST can improve APP695MT gene caused cell cycle arrest,promote cell to S phase transition,reduce apoptosis rate in PC12 cells,and have a protective effect on transfected cells.
