CAJ | 학술논문

目的：探讨自噬在低温治疗心肺脑复苏保护机制中对凋亡影响的研究。方法96只雄性新西兰家兔随机分为4组，空白对照（ Sham）组、复苏对照（ NT）组、低温治疗（ PATH）组及自噬抑制剂LY294002（LY294002）组。除Sham组每组再分为3个亚组，即自主循环恢复（ROSC）后4、9、24 h组，除Sham组为6只家兔外，另3组各时间点均为10只。模型采用经右心室致颤4 min后复苏模型，PATH组在ROSC后立即予低温干预，目标温度34℃，维持4 h；LY294002组用兔脑立体定位仪在复苏前20 min给予脑室内注射溶解于二甲亚砜的LY294002（1畅4 mg／kg ），并在复苏后予低温治疗。应用Western blot 方法测定脑组织 Bcl －2、Caspase －9、Caspase-3以及LC3－Ⅱ和Beclin－1的表达。结果①NT组ROSC后9、24 h Bcl－2、Caspase －9、Caspase －3、LC3－Ⅱ和Beclin－1表达较Sham组明显增加（P＜0．05），且NT组24 h较9 h明显增加（P＜0畅05）；②PATH组ROSC后9、24 h Bcl－2、Caspase－9、Caspase－3较NT组相应时间点减少（ P＜0．05），LC3－Ⅱ和Beclin－1较NT组相应时间点增加（P＜0．05）；③LY294002组ROSC后9、24 h Bcl－2、Caspase－9、Caspase－3较PATH组相应时间点增加（P＜0．05），LC3－Ⅱ和Beclin－1较PATH组相应时间点减少（ P＜0．05）。结论低温治疗通过抑制凋亡减轻心肺复苏后脑水肿，其机制可能是增加自噬调节引起。
목적：탐토자서재저온치료심폐뇌복소보호궤제중대조망영향적연구。방법96지웅성신서란가토수궤분위4조，공백대조（ Sham）조、복소대조（ NT）조、저온치료（ PATH）조급자서억제제LY294002（LY294002）조。제Sham조매조재분위3개아조，즉자주순배회복（ROSC）후4、9、24 h조，제Sham조위6지가토외，령3조각시간점균위10지。모형채용경우심실치전4 min후복소모형，PATH조재ROSC후립즉여저온간예，목표온도34℃，유지4 h；LY294002조용토뇌입체정위의재복소전20 min급여뇌실내주사용해우이갑아풍적LY294002（1창4 mg／kg ），병재복소후여저온치료。응용Western blot 방법측정뇌조직 Bcl －2、Caspase －9、Caspase-3이급LC3－Ⅱ화Beclin－1적표체。결과①NT조ROSC후9、24 h Bcl－2、Caspase －9、Caspase －3、LC3－Ⅱ화Beclin－1표체교Sham조명현증가（P＜0．05），차NT조24 h교9 h명현증가（P＜0창05）；②PATH조ROSC후9、24 h Bcl－2、Caspase－9、Caspase－3교NT조상응시간점감소（ P＜0．05），LC3－Ⅱ화Beclin－1교NT조상응시간점증가（P＜0．05）；③LY294002조ROSC후9、24 h Bcl－2、Caspase－9、Caspase－3교PATH조상응시간점증가（P＜0．05），LC3－Ⅱ화Beclin－1교PATH조상응시간점감소（ P＜0．05）。결론저온치료통과억제조망감경심폐복소후뇌수종，기궤제가능시증가자서조절인기。
Objective To investigate the protective role of hypothermia treatment for cardiopulmonary cerebral resuscitation and whether autophagy is involved in the protective mechanism . Methods Ninety-six male New Zealand rabbits were randomly divided into 4 groups:a sham group, a normothermic treatment group ( NT group ) , a post -arrest therapeutic hypothermia treatment group (PATH group) and an autophagy-suppressive LY294002 group(LY294002 group).Aside from a sham group of 6 rabbits, each group was divided into 3 sub -groups of 10 rabbits: 4, 6 or 24 hours after return of spontaneous circulation ( ROSC) .The NT group had only a right ventricular fibrillation for 4 minutes and was allowed to recover to ROSC .In the PATH group , the hypothermia intervention occurred immediately after ROSC.The target temperature was 34 ℃ and was maintained for 4 hours.In LY294002 group, an injection of LY294002 (1.4 mg/kg), which dissolves in dimethyl sulfoxide , into the rabbit's cerebral ventricle was made 20 minutes before ROSC occurred .The correct insertion location was determined by using a rabbit stereotaxic apparatus .Hypothermia intervention was also applied , occurring immediately after ROSC .The Western -blotting method is applied to determine the brain tissue Bcl-2, Caspase-9, Caspase-3 and LC3-Ⅱand Beclin-1 expression.Results For the NT group both 9 and 24 hours after ROSC all results significantly increased over the sham group ( P <0.05 ) , and within the NT group the 24 -hour subgroup was obviously increased over the 9 -hour subgroup(P<0.05).For the PATH group both 9 and 24 hours after ROSC Bcl -2, Caspase -9, Caspase-3 results decreased compared to the corresponding time point in the NT group (P<0.05), whereas the LC3-Ⅱand Beclin-1 results increased (P<0.05).For the LY294002 group both 9 and 24 hours after ROSC Bcl-2, Caspase-9, Caspase-3 results increased compared to the corresponding time point in the PATH group (P<0.05), whereas the LC3-Ⅱand Beclin-1 results decreased (P<0.05 ) .Conclusion Hypothermia treatment reduces brain edema after cardiopulmonary resuscitation by inhibiting apoptosis , the mechanism may be related to increased autophagy adjustment .