中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2012年
7期
500-504
,共5页
姜帅%梁直厚%魏桂荣%刘郁东
薑帥%樑直厚%魏桂榮%劉鬱東
강수%량직후%위계영%류욱동
阿尔茨海默病%β连环素%tau蛋白质类%地塞米松%糖原合成酶激酶3
阿爾茨海默病%β連環素%tau蛋白質類%地塞米鬆%糖原閤成酶激酶3
아이자해묵병%β련배소%tau단백질류%지새미송%당원합성매격매3
Alzheimer disease%beta Catenin%tau Proteins%Dexamethasone%Glycogen synthase kinase 3
目的 探讨Wnt/β-连环蛋白(catenin)信号通路在糖皮质激素(glucocorticoids,GC)诱导的阿尔茨海默病样病变中的信号调节机制.方法 地塞米松(dexamethasone,DEX)分别处理转染人tau441的人胚肾293细胞(HEK293/tau)和野生型的人胚肾293细胞(HEK293/wt),采用细胞计数试剂盒( CCK-8)法检测DEX对细胞活力的影响,Western blot研究两组细胞tau蛋白磷酸化(p-T205,Tau-1)、β-catenin、p-β-catenin、B细胞淋巴瘤/白血病-2蛋白(Bcl-2)及其上游激酶糖原合成激酶-3β(GSK-3β)、ps9-GSK-3β水平的变化,并加用GSK-3β的抑制剂氯化锂(LiCI),观察其对相关蛋白相应的逆转作用.结果 CCK-8细胞活力检测结果显示,1μmol/L DEX处理细胞48h,HEK293/wt组细胞活力(存活率)下降到95.5％±3.2％,HEK293/tau组下降到77.8％±4.4％,两者差异有统计学意义(t =6.60,P＜0.05)；Western blot结果显示,1μmoL/L DEX处理两组细胞48 h,使得HEK293/tau细胞ps9-GSK3β、Tau -I、β-catenin、Bcl-2水平分别下降到对照组的47.8％±10.4％、53.9％±11.7％、50.9％±7.6％、48.4％±6.5％,差异均有统计学意义(t=7.01、3.86、7.09、7.30,均P＜0.05),而p-T205、p-β-catenin相对磷酸化水平分别增加到对照组的180.5％±22.2％、201.3％±27.6％,差异均有统计学意义(t=5.51、5.27,均P＜0.05);LiCI可以相应逆转上述改变.结论 在人tau存在的前提下,GC通过抑制Wnt/β-catenin信号转导通路,促进了HEK293/tau细胞的阿尔茨海默病样病变.
目的 探討Wnt/β-連環蛋白(catenin)信號通路在糖皮質激素(glucocorticoids,GC)誘導的阿爾茨海默病樣病變中的信號調節機製.方法 地塞米鬆(dexamethasone,DEX)分彆處理轉染人tau441的人胚腎293細胞(HEK293/tau)和野生型的人胚腎293細胞(HEK293/wt),採用細胞計數試劑盒( CCK-8)法檢測DEX對細胞活力的影響,Western blot研究兩組細胞tau蛋白燐痠化(p-T205,Tau-1)、β-catenin、p-β-catenin、B細胞淋巴瘤/白血病-2蛋白(Bcl-2)及其上遊激酶糖原閤成激酶-3β(GSK-3β)、ps9-GSK-3β水平的變化,併加用GSK-3β的抑製劑氯化鋰(LiCI),觀察其對相關蛋白相應的逆轉作用.結果 CCK-8細胞活力檢測結果顯示,1μmol/L DEX處理細胞48h,HEK293/wt組細胞活力(存活率)下降到95.5％±3.2％,HEK293/tau組下降到77.8％±4.4％,兩者差異有統計學意義(t =6.60,P＜0.05)；Western blot結果顯示,1μmoL/L DEX處理兩組細胞48 h,使得HEK293/tau細胞ps9-GSK3β、Tau -I、β-catenin、Bcl-2水平分彆下降到對照組的47.8％±10.4％、53.9％±11.7％、50.9％±7.6％、48.4％±6.5％,差異均有統計學意義(t=7.01、3.86、7.09、7.30,均P＜0.05),而p-T205、p-β-catenin相對燐痠化水平分彆增加到對照組的180.5％±22.2％、201.3％±27.6％,差異均有統計學意義(t=5.51、5.27,均P＜0.05);LiCI可以相應逆轉上述改變.結論 在人tau存在的前提下,GC通過抑製Wnt/β-catenin信號轉導通路,促進瞭HEK293/tau細胞的阿爾茨海默病樣病變.
목적 탐토Wnt/β-련배단백(catenin)신호통로재당피질격소(glucocorticoids,GC)유도적아이자해묵병양병변중적신호조절궤제.방법 지새미송(dexamethasone,DEX)분별처리전염인tau441적인배신293세포(HEK293/tau)화야생형적인배신293세포(HEK293/wt),채용세포계수시제합( CCK-8)법검측DEX대세포활력적영향,Western blot연구량조세포tau단백린산화(p-T205,Tau-1)、β-catenin、p-β-catenin、B세포림파류/백혈병-2단백(Bcl-2)급기상유격매당원합성격매-3β(GSK-3β)、ps9-GSK-3β수평적변화,병가용GSK-3β적억제제록화리(LiCI),관찰기대상관단백상응적역전작용.결과 CCK-8세포활력검측결과현시,1μmol/L DEX처리세포48h,HEK293/wt조세포활력(존활솔)하강도95.5％±3.2％,HEK293/tau조하강도77.8％±4.4％,량자차이유통계학의의(t =6.60,P＜0.05)；Western blot결과현시,1μmoL/L DEX처리량조세포48 h,사득HEK293/tau세포ps9-GSK3β、Tau -I、β-catenin、Bcl-2수평분별하강도대조조적47.8％±10.4％、53.9％±11.7％、50.9％±7.6％、48.4％±6.5％,차이균유통계학의의(t=7.01、3.86、7.09、7.30,균P＜0.05),이p-T205、p-β-catenin상대린산화수평분별증가도대조조적180.5％±22.2％、201.3％±27.6％,차이균유통계학의의(t=5.51、5.27,균P＜0.05);LiCI가이상응역전상술개변.결론 재인tau존재적전제하,GC통과억제Wnt/β-catenin신호전도통로,촉진료HEK293/tau세포적아이자해묵병양병변.
Objective To investigate the role of Wnt/β-catenin pathway in the glucocorticoids (GC)-mediated Alzheimer' s disease-like pathological changes in vitro.Methods Human embryonic kidney 293 (HEK293/wt) cells stably transfected with the longest human tau (tau441,HEK293/tau) and wild-type HEK293 cells were employed to study the role of GC.Cell viabilities of the two cell lines were examined by cell counting kit-8 ( CCK-8 ).Levels of phosphorylated tau ( p-T205 ) and dephosphorylated tau (Tau-1),β-catenin,phosphorylated β-catenin (p-β-catenin),glycogen synthase kinase-3β (GSK-3β),phosphorylated GSK-3β at Ser9 (ps9-GSK-3β) and Bcl-2 were determined by Western blotting.Results Treatment with 1 μmol/L GC for 48 h decreased the viability of HEK293/wt and HEK293/tau cells to 95.5％ ±3.2％ and 77.8％ ± 4.4％ (t =6.60,P ＜ 0.05 ).Moreover,GC treatment decreased the levels of ps9-GSK-3β,Tau-1,β-catenin and Bcl-2 to 47.8％ ± 10.4％,53.9％ ± 11.7％,50.9％ ±7.6％,48.4％ ±6.5％ of control groups ( t =7.01,3.86,7.09,7.30,all P ＜ 0.05 ),and increased the relative levels of pT205,p-β-catenin to 180.5％ ± 22.2％,201.3 ％ ± 27.6％ of control groups (t =5.51,5.27,both P ＜0.05) only in HEK293/tau cells.Finally,LiCI efficiently prevented the above effects of GC in HEK293/tau cells.Conclusion GC may trigger Alzheimer' s disease-like pathological changes by inhibiting the Wnt/β-catenin pathway and these pathological processes seem to specifically depend on the presence of human tau.
