中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2009年
4期
368-371
,共4页
崔翔%陈彦青%刘荣国%李捷萌
崔翔%陳彥青%劉榮國%李捷萌
최상%진언청%류영국%리첩맹
二氮嗪%缺血预处理%JNK丝裂原活化蛋白激酶类%海马%神经元%细胞低氧%氧
二氮嗪%缺血預處理%JNK絲裂原活化蛋白激酶類%海馬%神經元%細胞低氧%氧
이담진%결혈예처리%JNK사렬원활화단백격매류%해마%신경원%세포저양%양
Diazoxide%Ischemic preconditioning%JNK mitogen-activated protein kinases%Hippocampus%Neurons%Cell hypoxia%Oxygen
目的 评价二氮嗪预处理对大鼠海马神经细胞缺氧复氧时c-Jun氨基末端激酶(JNK)表达的影响.方法 SD大鼠100只,出生<24 h,体重5～6 g,断头取海马组织,制备海马神经细胞悬液,海马神经细胞接种于96孔板或直径35 mm培养皿中,培养9～10 d后,随机分为4组,每组36孔或8皿:对照组(C组)、缺氧复氧组(A/R组)、二氮嗪30 μmol/L组(D1组)和二氮嗪100 μmol/L组(D2组).C组不行任何处理;A/R组置于特制的无菌密闭容器,容器中通人95%N2-5%CO2混合气体,流速0.2 L/min,4 h后放回原培养箱继续培养24 h;D1组和D2组在培养液中加入二氮嗪,终浓度分别为30、100 μmol/L,孵育1 h后用全量培养液洗脱,1次/d,连续3 d,随后处理同A/R组.于培养24 h时行免疫组化染色,观察神经细胞病理学结果 ,四甲基偶氮唑蓝比色法测定海马神经细胞活力,Westernblot法测定海马神经细胞Bcl-2、Bax和JNK的表达.结果 与C组比较,其余3组海马神经细胞Bcl-2、Bax和JNK表达均上调,活力降低(P<0.05或0.01);与A/R组比较,D2组海马神经细胞Bcl-2表达上调,Bax和JNK表达下调,活力升高(P<0.05或0.01),D1组上述指标差异无统计学意义(P>0.05).结论 二氮嗪预处理可能通过下调JNK的表达,抑制JNK信号通路,维持Bcl-2与Bax的平衡,减轻大鼠海马神经细胞缺氧复氧损伤.
目的 評價二氮嗪預處理對大鼠海馬神經細胞缺氧複氧時c-Jun氨基末耑激酶(JNK)錶達的影響.方法 SD大鼠100隻,齣生<24 h,體重5～6 g,斷頭取海馬組織,製備海馬神經細胞懸液,海馬神經細胞接種于96孔闆或直徑35 mm培養皿中,培養9～10 d後,隨機分為4組,每組36孔或8皿:對照組(C組)、缺氧複氧組(A/R組)、二氮嗪30 μmol/L組(D1組)和二氮嗪100 μmol/L組(D2組).C組不行任何處理;A/R組置于特製的無菌密閉容器,容器中通人95%N2-5%CO2混閤氣體,流速0.2 L/min,4 h後放迴原培養箱繼續培養24 h;D1組和D2組在培養液中加入二氮嗪,終濃度分彆為30、100 μmol/L,孵育1 h後用全量培養液洗脫,1次/d,連續3 d,隨後處理同A/R組.于培養24 h時行免疫組化染色,觀察神經細胞病理學結果 ,四甲基偶氮唑藍比色法測定海馬神經細胞活力,Westernblot法測定海馬神經細胞Bcl-2、Bax和JNK的錶達.結果 與C組比較,其餘3組海馬神經細胞Bcl-2、Bax和JNK錶達均上調,活力降低(P<0.05或0.01);與A/R組比較,D2組海馬神經細胞Bcl-2錶達上調,Bax和JNK錶達下調,活力升高(P<0.05或0.01),D1組上述指標差異無統計學意義(P>0.05).結論 二氮嗪預處理可能通過下調JNK的錶達,抑製JNK信號通路,維持Bcl-2與Bax的平衡,減輕大鼠海馬神經細胞缺氧複氧損傷.
목적 평개이담진예처리대대서해마신경세포결양복양시c-Jun안기말단격매(JNK)표체적영향.방법 SD대서100지,출생<24 h,체중5～6 g,단두취해마조직,제비해마신경세포현액,해마신경세포접충우96공판혹직경35 mm배양명중,배양9～10 d후,수궤분위4조,매조36공혹8명:대조조(C조)、결양복양조(A/R조)、이담진30 μmol/L조(D1조)화이담진100 μmol/L조(D2조).C조불행임하처리;A/R조치우특제적무균밀폐용기,용기중통인95%N2-5%CO2혼합기체,류속0.2 L/min,4 h후방회원배양상계속배양24 h;D1조화D2조재배양액중가입이담진,종농도분별위30、100 μmol/L,부육1 h후용전량배양액세탈,1차/d,련속3 d,수후처리동A/R조.우배양24 h시행면역조화염색,관찰신경세포병이학결과 ,사갑기우담서람비색법측정해마신경세포활력,Westernblot법측정해마신경세포Bcl-2、Bax화JNK적표체.결과 여C조비교,기여3조해마신경세포Bcl-2、Bax화JNK표체균상조,활력강저(P<0.05혹0.01);여A/R조비교,D2조해마신경세포Bcl-2표체상조,Bax화JNK표체하조,활력승고(P<0.05혹0.01),D1조상술지표차이무통계학의의(P>0.05).결론 이담진예처리가능통과하조JNK적표체,억제JNK신호통로,유지Bcl-2여Bax적평형,감경대서해마신경세포결양복양손상.
Objective To investigate the effects of diazoxide preconditioning on the expression of C-Jun N-terminal kinase (JNK) in primary cultured rat hippocampal neurons after anoxia-reoxygenation (A/R). Methods One hundred SD rats (<24 h after birth) weighing 5-6 g were decapitated. Their hippocampi were removed and cut into chunks. Hippocampal neurons were enzymatically isolated. After being cultured for 9-10 d, the hippocampal neurons were randomly divided into 4 groups: group Ⅰ control (group C);group Ⅱ A/R;group Ⅲ, Ⅳ preconditioning with diazoxide 30 and 100 μmol/L (group D1, D2). In group A/R, D1 and D2 the neurons were exposed to 95 % N2-5% CO2 for 4 h followed by 24 hreoxygenation. In group D1 and D2 the neurons were pretreated with diazoxide 30 and 100 μmol/L respectively for 1 h before A/R for 3 consecutive days. The viability of neurons was measured by MTT assay. The expression of Bcl-2, Bax and JNK proteins was detected by Western blot analysis. Results The viability of neurons was significant lower in group A/R, D, and D2 than in control group and was significantly higher in group D2 than in group A/R. The expression of Bcl-2, Bax and JNK proteins was significantly up-regulated in group A/R, D1 and D2 as compared with control group. The expression of Bcl-2was significantly higher while the Bax and JNK protein expression was significantly lower in group D2 than in group A/R. Conclusion Preconditioning with diazoxide 100 μmol/L can ameliorate A/R injury to hippocampal neurons by down-regulatlng Bax expression and up-regulating Bcl-2 expression through inhibition of JNK signaling transduction pathway.
