CAJ | 학술논문

目的评价七氟醚预处理对大鼠脊髓缺血再灌注损伤的影响及自噬在其中的作用.方法成年雄性SD大鼠45只,体重420～450 g,采用随机数字表法分为5组(n=9):对照组(Con组)、脊髓缺血再灌注组(I/R组)、七氟醚预处理组(Sevo组)、特异性自噬抑制剂3-甲基腺嘌呤组(3-MA组)和3-MA+七氟醚预处理组(3-MA+ Sevo组).I/R组胸主动脉球囊阻断+体循环低血压制备大鼠脊髓缺血再灌注模型,Sevo组于缺血前24h时吸入3.4％七氟醚2h,3-MA组和3-MA+ Sevo组分别于再灌注即刻和吸入七氟醚前15 min时鞘内注射20出3-MA(10 mmol/L).于再灌注24h时采用神经功能缺陷评分(NDS评分)法评价大鼠神经功能,随后处死取脊髓,Western blot法检测LC3B、Beclin 1、Bcl-2蛋白的表达水平.结果与Con组比较,I/R组脊髓LC3B、Beclin 1蛋白表达上调,Bcl-2蛋白表达下调,NDS评分升高(P＜0.05)；与I/R组比较,Sevo组、3-MA组和3-MA+ Sevo组脊髓LC3B、Beclin 1蛋白表达下调,Bcl-2蛋白表达上调,NDS评分降低(P＜0.05)；Sevo组、3-MA组和3-MA+ Sevo组各指标比较差异无统计学意义(P＞0.05).结论七氟醚预处理可减轻大鼠脊髓缺血再灌注损伤,其机制可能与上调Bcl-2,抑制自噬溶酶体途径,减轻自噬有关.
목적 평개칠불미예처리대대서척수결혈재관주손상적영향급자서재기중적작용.방법 성년웅성SD대서45지,체중420～450 g,채용수궤수자표법분위5조(n=9):대조조(Con조)、척수결혈재관주조(I/R조)、칠불미예처리조(Sevo조)、특이성자서억제제3-갑기선표령조(3-MA조)화3-MA+칠불미예처리조(3-MA+ Sevo조).I/R조흉주동맥구낭조단+체순배저혈압제비대서척수결혈재관주모형,Sevo조우결혈전24h시흡입3.4％칠불미2h,3-MA조화3-MA+ Sevo조분별우재관주즉각화흡입칠불미전15 min시초내주사20출3-MA(10 mmol/L).우재관주24h시채용신경공능결함평분(NDS평분)법평개대서신경공능,수후처사취척수,Western blot법검측LC3B、Beclin 1、Bcl-2단백적표체수평.결과 여Con조비교,I/R조척수LC3B、Beclin 1단백표체상조,Bcl-2단백표체하조,NDS평분승고(P＜0.05)；여I/R조비교,Sevo조、3-MA조화3-MA+ Sevo조척수LC3B、Beclin 1단백표체하조,Bcl-2단백표체상조,NDS평분강저(P＜0.05)；Sevo조、3-MA조화3-MA+ Sevo조각지표비교차이무통계학의의(P＞0.05).결론 칠불미예처리가감경대서척수결혈재관주손상,기궤제가능여상조Bcl-2,억제자서용매체도경,감경자서유관.
Objective To evaluate the effect of sevoflurane preconditioning on spinal cord ischemia-reperfusion (I/R) injury in rats and the role of autophagy.Methods Forty-five healthy adult male Sprague-Dawley rats,weighing 420-450 g,were equally and randomly divided into 5 groups:control group (Con group),spinal cord IIR group (group I/R),sevoflurane preconditioning group (group Sevo),a specific autophagy inhibitor 3-methyladenine (3-MA) group (group 3-MA) and 3-MA + sevoflurane preconditioning group (group 3-MA + Sevo).The animals were anesthetized with intraperitoneal 4％ chloral hydrate 0.1 mg/kg.Spinal cord ischemia was induced by 10 min occlusion of the thoracic aorta combined with controlled hypotension (MAP =40 mm Hg).In group Sevo,3.4％ sevoflurane was inhaled for 2 h starting from 24 h before ischemia.20 μl 3-MA (10 mmol/L) was injected intrathecally immediately after onset of reperfusion and at 15 min before sevoflurane inhalation,in 3MA and 3-MA + Sevo groups,respectively.Neurological deficit was scored at 24 h of reperfusion.The rats were then sacrificed and the spinal cord was removed for detection of the expression of LC-3B,Beclin 1 and Bcl-2.Results Compared with Con group,the expression of LC-3B and Beclin 1 was significantly up-regulated,the expression of Bcl-2 was down-regulated,and neurological deficit scores were increased in I/R group (P ＜ 0.05).Compared with group I/R,the expression of LC-3B and Beclin 1 was significantly down-regulated,the expression of Bcl-2 was up-regulated,and neurological deficit scores were decreased in groups Sevo,3-MA and 3-MA + Sevo (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups Sevo,3-MA and 3-MA + Sevo (P ＞ 0.05).Conclusion Sevoflurane preconditioning can reduce spinal cord I/R injury and the underlying mechanism may be related to up-regulation of the expression of Bcl-2,inhibition of autophagy-lysosomal pathway,and reduction of autophagy in rats.