中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
7期
1463-1465
,共3页
王正锋%王瑞华%刘献志%翟广%张水军
王正鋒%王瑞華%劉獻誌%翟廣%張水軍
왕정봉%왕서화%류헌지%적엄%장수군
脊髓缺血性损伤%小泛素化调节因子%神经保护
脊髓缺血性損傷%小汎素化調節因子%神經保護
척수결혈성손상%소범소화조절인자%신경보호
Spinal cord ischemia injury%Small ubiquitin-like modifier%Neuroprotection
目的 观察脊髓缺血性损伤是否激活小泛素化调节因子(SUMO)以及SUMO结合蛋白在缺血刺激后的亚细胞定位.方法 制备小鼠脊髓缺血模型,用Western blot法检验脊髓缺血性损伤引起的应激反应,再分别用Western blot和免疫荧光染色法观察缺血脊髓节段中SUMO结合蛋白的表达和亚细胞定位.结果 在脊髓缺血性损伤模型中,应激反应被激活,真核细胞起始因子2(eIF2)、细胞外信号调节激酶(ERK) 1/2和蛋白激酶B(Akt)的磷酸化水平明显增高.夹闭8、10、12 min组小鼠损伤脊髓节段内SUMO1的表达量分别是对照脊髓节段的(1.15±0.09)、(1.24±0.13)、(1.01 +0.99)倍,差异无统计学意义(P＞0.05),SUMO2/3分别是对照脊髓节段的(5.87±0.31)、(5.40±0.36)、(4.31±0.28)倍,差异有统计学意义(P＜0.01).免疫荧光染色结果显示损伤脊髓节段SUMO2/3结合蛋白增多并发生明显的核聚集.结论 SUMO2/3在脊髓缺血性损伤的应激反应中起到重要的作用,可能是内源性神经保护途径的关键分子.
目的 觀察脊髓缺血性損傷是否激活小汎素化調節因子(SUMO)以及SUMO結閤蛋白在缺血刺激後的亞細胞定位.方法 製備小鼠脊髓缺血模型,用Western blot法檢驗脊髓缺血性損傷引起的應激反應,再分彆用Western blot和免疫熒光染色法觀察缺血脊髓節段中SUMO結閤蛋白的錶達和亞細胞定位.結果 在脊髓缺血性損傷模型中,應激反應被激活,真覈細胞起始因子2(eIF2)、細胞外信號調節激酶(ERK) 1/2和蛋白激酶B(Akt)的燐痠化水平明顯增高.夾閉8、10、12 min組小鼠損傷脊髓節段內SUMO1的錶達量分彆是對照脊髓節段的(1.15±0.09)、(1.24±0.13)、(1.01 +0.99)倍,差異無統計學意義(P＞0.05),SUMO2/3分彆是對照脊髓節段的(5.87±0.31)、(5.40±0.36)、(4.31±0.28)倍,差異有統計學意義(P＜0.01).免疫熒光染色結果顯示損傷脊髓節段SUMO2/3結閤蛋白增多併髮生明顯的覈聚集.結論 SUMO2/3在脊髓缺血性損傷的應激反應中起到重要的作用,可能是內源性神經保護途徑的關鍵分子.
목적 관찰척수결혈성손상시부격활소범소화조절인자(SUMO)이급SUMO결합단백재결혈자격후적아세포정위.방법 제비소서척수결혈모형,용Western blot법검험척수결혈성손상인기적응격반응,재분별용Western blot화면역형광염색법관찰결혈척수절단중SUMO결합단백적표체화아세포정위.결과 재척수결혈성손상모형중,응격반응피격활,진핵세포기시인자2(eIF2)、세포외신호조절격매(ERK) 1/2화단백격매B(Akt)적린산화수평명현증고.협폐8、10、12 min조소서손상척수절단내SUMO1적표체량분별시대조척수절단적(1.15±0.09)、(1.24±0.13)、(1.01 +0.99)배,차이무통계학의의(P＞0.05),SUMO2/3분별시대조척수절단적(5.87±0.31)、(5.40±0.36)、(4.31±0.28)배,차이유통계학의의(P＜0.01).면역형광염색결과현시손상척수절단SUMO2/3결합단백증다병발생명현적핵취집.결론 SUMO2/3재척수결혈성손상적응격반응중기도중요적작용,가능시내원성신경보호도경적관건분자.
Objective To determine whether spinal cord ischemia induces small ubiquitin-like modifier (SUMO) conjugation and to observe the subcelluar location of SUMO conjugation.Methods Activation of stress response pathways after spinal cord ischemia was verified by Western blotting analysis in mice spinal ischemia model.Levels and subcellular localizations of SUMO-conjugated proteins in spinal cords were evaluated by Western blotting analysis and immunol fluorescene staining,respectively.Results Following transient spinal cord ischemia,stress responses were activated as indicated by increased phosphorylation of eukaryotic initiation factor 2 (eIF2),extracellular signal-regulated kinase (ERK)1/2,and protein kinase B (Akt).Leves of SUMO1-conjugated proteins were not change significantly after 8,10,or 12 min ischemia,1.15 ±0.09,1.24 ± 0.13,1.01 ± 0.99,respectively (P ＞ 0.05).Levels of SUMO2/3conjugated proteins were markedly increased:5.87 ±0.31,5.40 ±0.36,4.31 ±0.28,respectively (P ＜0.01).The result of immunol fluorescene staining showed that SUMO2/3-conjugation proteins increased markedly and nulei accumulation occurred.Conclusion SUMO2/3 maybe a key molecular in internal neuroprotective pathway which played an important role in the stress response of spinal cord ischemia.
