CAJ | 학술논문

目的：探讨异氟醚麻醉对新生大鼠海马组织的 p38丝裂原活化蛋白激酶通路（mitogen-activated protein kinase， MAPK）激活的影响，以及 p38 MAPK 信号通路对异氟醚诱导海马神经细胞凋亡的影响。方法48只出生后7 d 的新生大鼠随机分为 DMSO 对照组（Air ＋DMSO 组）、p38 MAPK 抑制剂 SB203580对照组（Air ＋SB20组）、异氟醚＋DMSO 组（Iso ＋DMSO 组）和异氟醚＋SB203580组（Iso ＋SB20组）。对照组吸入空气，异氟醚组吸入体积分数为0.011异氟醚4 h。麻醉前30 min，分别侧脑室注射 SB203580（20 nmol）或者体积分数为0.1 DMSO 5μl。麻醉结束后6 h，部分幼鼠灌注取脑，TUNEL 荧光染色检测幼鼠脑海马 CA1区神经细胞凋亡（n ＝6）；部分幼鼠取新鲜脑海马，Western blot 法检测磷酸化p38（phospho-p38，p-p38）、p38、cleaved caspase-3、磷酸化NF-κB（phospho-NF-κB，p-NF-κB）、Bcl-2、Bax 等蛋白表达的变化（n ＝6）。结果Iso ＋DMSO 组海马 CA1区 TUNEL 阳性细胞数比 Air ＋DMSO 组增加了4.8倍（P ＜0.01），与 Iso＋DMSO 组相比，Iso ＋SB20组海马 CA1区 TUNEL 阳性细胞数降低了约3／5（P ＜0.01）；Iso ＋DMSO 组海马 cleaved caspase-3蛋白表达较 Air ＋DMSO 组表达增加（P ＝0.003）， Iso ＋SB20组减少了异氟醚引起的海马 cleaved caspase-3增加（P ＝0.007）；与 Air ＋DMSO 组比较，异氟醚增加了 p-p38及其下游 p-NF-κB 的表达，增加了 Bax 的表达，降低了 Bcl-2的表达；而 p38抑制剂 SB203580则减少了异氟醚引起的 p-p38、p-NF-κB 和 Bax 的增加，增加了 Bcl-2的表达。结论异氟醚通过激活 p38 MAPK 信号通路诱导发育期大鼠海马神经细胞凋亡。
목적：탐토이불미마취대신생대서해마조직적 p38사렬원활화단백격매통로（mitogen-activated protein kinase， MAPK）격활적영향，이급 p38 MAPK 신호통로대이불미유도해마신경세포조망적영향。방법48지출생후7 d 적신생대서수궤분위 DMSO 대조조（Air ＋DMSO 조）、p38 MAPK 억제제 SB203580대조조（Air ＋SB20조）、이불미＋DMSO 조（Iso ＋DMSO 조）화이불미＋SB203580조（Iso ＋SB20조）。대조조흡입공기，이불미조흡입체적분수위0.011이불미4 h。마취전30 min，분별측뇌실주사 SB203580（20 nmol）혹자체적분수위0.1 DMSO 5μl。마취결속후6 h，부분유서관주취뇌，TUNEL 형광염색검측유서뇌해마 CA1구신경세포조망（n ＝6）；부분유서취신선뇌해마，Western blot 법검측린산화p38（phospho-p38，p-p38）、p38、cleaved caspase-3、린산화NF-κB（phospho-NF-κB，p-NF-κB）、Bcl-2、Bax 등단백표체적변화（n ＝6）。결과Iso ＋DMSO 조해마 CA1구 TUNEL 양성세포수비 Air ＋DMSO 조증가료4.8배（P ＜0.01），여 Iso＋DMSO 조상비，Iso ＋SB20조해마 CA1구 TUNEL 양성세포수강저료약3／5（P ＜0.01）；Iso ＋DMSO 조해마 cleaved caspase-3단백표체교 Air ＋DMSO 조표체증가（P ＝0.003）， Iso ＋SB20조감소료이불미인기적해마 cleaved caspase-3증가（P ＝0.007）；여 Air ＋DMSO 조비교，이불미증가료 p-p38급기하유 p-NF-κB 적표체，증가료 Bax 적표체，강저료 Bcl-2적표체；이 p38억제제 SB203580칙감소료이불미인기적 p-p38、p-NF-κB 화 Bax 적증가，증가료 Bcl-2적표체。결론이불미통과격활 p38 MAPK 신호통로유도발육기대서해마신경세포조망。
Aim To investigate the effect of isoflurane on the phosphorylation of p38 mitogen-activated protein kinase (MAPK)in the hippocampus of neonatal rats, and the effect of p38 MAPK pathway on isoflurane-in-duced neuronal apoptosis.Methods Forty-eight neo-natal rats on postnatal day 7 were assigned randomly into four groups:DMSO group (group Air +DMSO), p38 MAPK inhibitor SB203580 group (group Air +SB20 ),isoflurane +DMSO group (group Iso +DM-SO),and isoflurane +SB203580 group (group Iso +SB20 ).Rats were exposed to air or isoflurane (volume fraction of 0.01 1 )for 4h.The p38 inhibitor SB203580 (20 nmol)or DMSO (volume fraction of 0.1 )5μl was intraventricularly administered 30 min before the expo-sure.The brains of some rats in each group were per-fused and embedded by paraffin 6h after the exposure. Neuronal apoptosis in the hippocampal CA1 area was detected by terminal deoxyribonucleotide transferase-mediated dUTP nick end labeling (TUNEL)(n =6). The hippocampal tissues of the other rats in each group were dissected 6h after the exposure,and the protein expressions of phospho-p38 (p-p38 ),p38,cleaved caspase-3,phospho-NF-κB (p-NF-κB ),Bcl-2 and Bax were detected by Westem blot (n =6).Results The number of TUNEL positive cells in the hippocam-pal CA1 region in group Iso +DMSO increased by 4.8 fold compared with that in group Air +DMSO (P <0.01 ),while the number of TUNEL positive cells in group Iso +SB20 decreased by 3 /5 compared with that in group Iso +DMSO (P <0.01 ).The protein expres-sion of cleaved caspase-3 in group Iso +DMSO signifi-cantly increasd (P =0.003)compared to that in group Air +DMSO,which was significantly decreasd in group Iso +SB20 (P =0.007 ).In addition,isoflurane also increased the protein expression of p-p38,p-NF-κB and Bax,decreased the level of Bcl-2,and reduced the ratio of Bcl-2 /Bax compared with control animals (P <0.01 ,P =0.004,P <0.01 ,P <0.01 ,P <0.01 ,respectively).Howerver,SB203580 partly at-tenuated the isoflurane-induced protein change above. Conclusion Isoflurane induces neuroapoptosis in neo-natal rat hippocampus by the activation of p38 MAPK pathway.