CAJ | 학술논문

目的探讨二氮嗪(DZ)预处理能否通过上调Akt信号增强Bcl-2表达、抑制Bax表达发挥抗凋亡作用.方法离体培养9～10 d SD大鼠海马神经元分为正常对照组(A组)、缺氧组(B组)、缺氧+DZ 100 μmol/L组(c组)、缺氧+DZ 100 μmol/L+5-羟癸酸100 μmol/L组(D组)、缺氧+DZ 100 μmol/L+LY294002 50 μmol/L组(E组),自缺氧前2 d开始,神经元接受DZ预处理,每天1次,每次1 h.每次实验,每组16孔或2皿细胞,实验重复3次.比较缺氧4 h复氧48 h各组海马神经元的活力、凋亡率、Akt、Bcl-2和Bax蛋白的表达程度.结果缺氧复氧后C组吸光度值较B、D、E组显著增高(P＜0.05);其他缺氧各组间比较,差异无统计学意义(P＞0.05).C组凋亡率较B、D、E组显著减低(P＜0.05).C组Akt、Bcl-2表达较B、D、E组强烈(P＜0.05),Bax表达则减弱(P＜0.05).B、D、E组问比较,差异无统计学意义(P＞0.05).结论 DZ可经Akt信号通路上调Bcl-2/Bax蛋白比值而抑制缺氧复氧损伤大鼠海马神经元的凋亡.
목적 탐토이담진(DZ)예처리능부통과상조Akt신호증강Bcl-2표체、억제Bax표체발휘항조망작용.방법 리체배양9～10 d SD대서해마신경원분위정상대조조(A조)、결양조(B조)、결양+DZ 100 μmol/L조(c조)、결양+DZ 100 μmol/L+5-간계산100 μmol/L조(D조)、결양+DZ 100 μmol/L+LY294002 50 μmol/L조(E조),자결양전2 d개시,신경원접수DZ예처리,매천1차,매차1 h.매차실험,매조16공혹2명세포,실험중복3차.비교결양4 h복양48 h각조해마신경원적활력、조망솔、Akt、Bcl-2화Bax단백적표체정도.결과 결양복양후C조흡광도치교B、D、E조현저증고(P＜0.05);기타결양각조간비교,차이무통계학의의(P＞0.05).C조조망솔교B、D、E조현저감저(P＜0.05).C조Akt、Bcl-2표체교B、D、E조강렬(P＜0.05),Bax표체칙감약(P＜0.05).B、D、E조문비교,차이무통계학의의(P＞0.05).결론 DZ가경Akt신호통로상조Bcl-2/Bax단백비치이억제결양복양손상대서해마신경원적조망.
Objective Investigate whether preconditioning with diazoxide(DZ),a mitochondrial ATP-sensitive potassium channel opener,could enhance Akt protein to up-regulate Bcl-2/Bax protein ratio against apoptosis.Methods Cultured for 9-10 d in vitro,the hippocampat neurons of Sprague-Dawley rats were assigned to the following 5 groups randomly:Control(Group A),Anoxia(Group B),Anoxia+DZ100 μmoL/L(Group C),Anoxia+DZ100 μmol/L+5-hydroxydecanoate(Group D),Anoxia+DZ100 μmol/L+LY294002 50 μmol/L(Group E).Prior to oxygen deprivation,the hippocampal neurons were treated with DZ for 1 h per day and this treatment was persisted for 3 d.Each experiment was repeated for three times and each group contained 16 wells or 2 dishes of neurons for each time.Thereafter,neurons were derived of oxygen for 4 h.At 48 h of reoxygenation the neuronal optical density(A)were measured by MTT method,while the apoptotic rates were assayed by annexin V-FITC staining.The expressions of Akt,Bcl-2 and Bax proteins were detected and evaluated by Western blot.Results Compared with other pretreatment,DZ 100 μmoL/L led to the elevation of A,whereas promoted the decrease of apoptotic rate(P ＜ 0.05).Compared with those in other anoxic groups,the expressions of Akt protein and Bcl-2 protein in Group C were increased significantly(P ＜0.05),whereas the Bax density were reduced significantly(P ＜ 0.05).Preceding administration of 100 μmol/L DZ took protective effects on the neurons induced by anoxia-reoxygenation.Conclusions DZ 100 μmol/L increased Akt protein to up-regulate Bcl-2/Bax protein ratio against apoptosis induced by anoxia-reoxygenation.