CAJ | 학술논문

目的：研究蛋白激酶信号级联蛋白激酶B（Akt）在大鼠脊髓损伤（SCI）后的表达变化，以及对SCI后运动功能恢复的影响，从而为SCI的修复提供分子学机制依据。方法采用改良 Allen′s 打击器制成大鼠SCI模型。大鼠分为3组：假手术组（Sham组）、干预组（Int组）、对照组（Con组）。分别于术后1、7、14 d采用免疫组织化学、蛋白免疫印迹法（Western blot）和实时荧光定量-聚合酶链反应（RT-PCR）检测Akt mRNA和蛋白的表达，并采用Basso-Beattie-Bresnahan（BBB）运动功能评分评价大鼠SCI后运动恢复情况。结果 Sham组在Akt蛋白和mRNA水平上呈低水平表达，SCI后Akt的表达水平明显提高；Akt上游磷脂酰肌醇3-激酶（PI3K）的抑制剂LY294002可明显抑制Akt的表达，并阻止SCI后自发性运动功能恢复。结论 SCI后激活的蛋白激酶信号Akt可能是参与SCI后神经保护和修复的重要干预环节。
목적：연구단백격매신호급련단백격매B（Akt）재대서척수손상（SCI）후적표체변화，이급대SCI후운동공능회복적영향，종이위SCI적수복제공분자학궤제의거。방법채용개량 Allen′s 타격기제성대서SCI모형。대서분위3조：가수술조（Sham조）、간예조（Int조）、대조조（Con조）。분별우술후1、7、14 d채용면역조직화학、단백면역인적법（Western blot）화실시형광정량-취합매련반응（RT-PCR）검측Akt mRNA화단백적표체，병채용Basso-Beattie-Bresnahan（BBB）운동공능평분평개대서SCI후운동회복정황。결과 Sham조재Akt단백화mRNA수평상정저수평표체，SCI후Akt적표체수평명현제고；Akt상유린지선기순3-격매（PI3K）적억제제LY294002가명현억제Akt적표체，병조지SCI후자발성운동공능회복。결론 SCI후격활적단백격매신호Akt가능시삼여SCI후신경보호화수복적중요간예배절。
Objective To examine the expression of protein kinase signaling cascade protein kinase B (Akt)-which plays an im-portant role in a number of key biological functions in cellular processes after spinal cord injury (SCI) in rats ,and its effects on SCI induced motor function defects ,and to provide the molecular mechanism for repairing SCI .Methods SCI was produced by extra-dural contusion using modified Allen′s stall with damage energy .The rats were divided into three groups :sham-operated group (Sham) ,interference group (Int) ,and control group (Con) .Using immunostaining studies ,Western blot analyses and real-time qualitative RT-PCR analyses ,we detected the changes of Akt expression at the protein and mRNA level in spinal cord tissues at 1 , 7 ,and 14 d after surgery ,and evaluated the presence and extent of neurological impairment after SCI by the BBB locomotor rating scale .Results The sham operated groups exhibited low expression of the Akt signaling at the protein and mRNA level ,and the ex-pressions increased following SCI .Compared to the animals in the sham operated groups ,prominently elevated level of Akt signaling was detected in the injured spinal cords of SCI group .LY294002 ,an inhibitor of phosphatidylinositol 3-kinase (PI3K) that initiates Akt phosphorylation ,prominently inhibited the expression of Akt produced by SCI and the spontaneous recovery of motor function after SCI .Conclusion Activated protein kinase signaling cascade Akt might be the important intervention aspects of involving in neuroprotective and repair process after SCI .