盐酸羟考酮预先给药对大鼠脊髓缺血再灌注损伤时细胞凋亡的影响  

Effect of oxycodone hydrochloride pretreatment on cell apoptosis during spinal cord ischemia-reperfusion in rats

作　　者：赵加[1] 张伟[1] 尹若峰[2] 张伯寅[2] Zhao Jia;Zhang Wei;Yin Ruofeng;Zhang Boyin(Department of Anesthesiology,China-Japan Union Hospital of Jilin University,Changchun 130033,China;Department of Orthopedics,China-Japan Union Hospital of Jilin University,Changchun 130033,China)

机构地区：[1]吉林大学中日联谊医院麻醉科,长春130033 [2]吉林大学中日联谊医院骨科,长春130033

出　　处：《中华实验外科杂志》2020年第6期1049-1051,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81701225);吉林省教育厅科研规划项目(JJKH20201056KJ);吉林省科技厅省自然基金(20200201436JC);吉林省科技厅优青项目(20190103077JH)。

摘　　要：目的观察盐酸羟考酮预先给药对脊髓缺血再灌注(I/R)损伤时细胞凋亡的影响。方法2019年8月至2019年12月,选取无特定病原体(SPF)级健康雄性大鼠36只,体重200~270 g,来源于吉林大学动物实验中心,采用随机数字表达法分为3组(n=12):假手术组(Sham组)、脊髓缺血再灌注(I/R组)和盐酸羟考酮组(Oxy组)。I/R组夹闭腹主动脉制备脊髓缺血再灌注模型;Oxy组大鼠于缺血前15 min静脉注射盐酸羟考酮0.5 mg/kg;Sham组和I/R组给予同等容量的生理盐水。灌注后6、12、24 h评价后肢功能运动评分。再灌注24 h时取脊髓组织,光镜下观察脊髓前角运动神经元病理学结果,原位缺口末端标记法(TUNEL)法进行细胞凋亡率测定,采用免疫组织化学法测定B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)和半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3蛋白的表达水平。应用SPSS 21.0统计软件分析,组间比较采用单因素方差分析。结果与Sham组(7.00、7.00、7.00分)比较,I/R组和Oxy组在再灌注6 h[(2.67±0.52)、(4.33±0.52)分]、12 h[(3.33±0.52)、(4.83±0.41)分]、24 h[(4.17±0.75)、(5.67±0.52)分]时后肢运动功能评分降低(F=161.250、141.150、43.400,P<0.05),脊髓前角运动神经元病理学损伤较重,差异有统计学意义;与Sham组[(3.42±0.90)%]比较,I/R组[(29.58±6.40)%]和Oxy组[(9.42±2.11)%]细胞凋亡率升高(F=146.260,P<0.05),差异有统计学意义;与Sham组(6.83±1.27、7.08±1.56、4.58±1.08)比较,I/R组和Oxy组脊髓组织中bcl-2(36.75±5.17、36.75±5.17)、bax(50.92±7.03、29.42±5.37)和Caspase-3蛋白(45.25±5.22、30.58±3.40)的表达水平升高(F=308.000、214.530、381.670,P<0.05),差异有统计学意义。与I/R组比较,Oxy组在再灌注6 h(2.67±0.52、4.33±0.52)、12 h(3.33±0.52、4.83±0.41)、24 h(4.17±0.75、5.67±0.52)时后肢运动功能评分升高(F=152.460、126.480、46.750,P<0.05),脊髓前角运动神经元病理学损伤较轻,细胞凋亡率[(29.58±6.40)%、(Objective To evaluate the effect of oxycodone hydrochloride(Oxy)pretreatment on cell apoptosis during spinal cord ischemia/reperfusion(I/R)in rats.Methods From August to December 2019,36 specific pathogen free(SPF)healthy male rats,weighing 200-270 g,purchased from the Animal Experiment Center of Jilin University,were randomly divided into 3 groups(n=12 each):sham operation group(sham group),spinal cord I/R group(I/R group)and Oxy group.Spinal cord I/R model was induced by temporary occlusion of the abdominal aorta in the I/R group and Oxy group.Oxy(0.5 mg/kg)was intravenously injected at 15 min before ischemia in the Oxy group.The equal volume of normal saline was given instead in the sham group and I/R group.The motor nerve functions of the hindlimb in rats were assessed at 6,12 and 24 h after reperfusion.Spinal cords were removed at 24 h after the reperfusion.Pathological changes of spinal cord anterior horn motor neurons were observed with a light microscope.The apoptosis ratio was calculated by TdT-mediated dUTP nick end labeling(TUNEL),and the expression of the B cell lymphoma/leukemia-2(bcl-2),bcl-2 associated X protein(bax)and cysteinyl aspartate-specific protease(Caspase)-3 was detected by immunohistochemistry.Statistic analysis of data was done using the statistical product and service solution 21.0 software.Results Compared with sham group(7.00,7.00,7.00 score),the motor nerve functions of the hindlimb in rats at 6 h[(2.67±0.52),(4.33±0.52)score],12 h[(3.33±0.52),(4.83±0.41)score]and 24 h[(4.17±0.75),(5.67±0.52)score]after reperfusion were significantly decreased(F=161.250,141.150,43.400,P<0.05).The pathological injury of spinal cord anterior horn motor neurons was more serious.The apoptosis ratio was significantly higher in the L/R group[(29.58±6.40)%]and Oxy group[(9.42±2.11)%,F=146.260,P<0.05]than in the sham group[(3.42±0.90)%].Compared with sham group(6.83±1.27,7.08±1.56,4.58±1.08),the expression levels of bcl-2(36.75±5.17,36.75±5.17),bax(50.92±7.03,29.42±5.37)and Caspase-3(45.25±

关 键 词：脊髓缺血 再灌注损伤 盐酸羟考酮 脱噬作用 

分 类 号：R744[医药卫生—神经病学与精神病学]