依达拉奉抗大鼠脊髓损伤后脊髓细胞凋亡的机制初探  

Preliminary mechanism of edaravone against cell apoptosis after spinal cord injury in rats

作　　者：王继权[1] 赵兴长 孙平[1] 李昊天[1] 褚鑫[1] 吕刚[1] 范仲凯[1] 

机构地区：[1]辽宁省辽宁医学院附属第一医院骨科,辽宁锦州121001

出　　处：《天津医药》2015年第9期988-991,I0004,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81272074);辽宁省高等学校优秀人才支持计划项目(LJQ2014091);辽宁医学院校长基金项目(QM2014011)

摘　　要：目的探讨依达拉奉(EDA)对大鼠脊髓损伤(SCI)后内质网应激(ERS)介导的细胞凋亡的影响。方法36只SD大鼠随机分为假手术(sham)组、SCI组、EDA组,每组12只。采用Allen’s法建立SCI模型,sham组仅行椎板切除术。术后Sham组和SCI组给予与EDA组等体积等频次生理盐水处理;EDA组在SCI模型建成以后予以EDA(10 mg/kg),每12 h腹腔注射给药1次。于术后3 d取脊髓,应用Western blot检测C/EBP同源蛋白(CHOP)、Cleaved caspase-12和Cleaved caspase-3的表达水平,免疫荧光技术检测脊髓组织中caspase-12、CHOP阳性细胞的比率,TUNEL法检测脊髓细胞凋亡水平。结果 SCI组较sham组CHOP、Cleaved caspase-12、Cleaved caspase-3的表达升高,Cleaved caspase-12、CHOP阳性细胞比率增加,脊髓组织细胞凋亡比率增加(均P<0.01)。EDA组较SCI组CHOP、Cleaved caspase-12和Cleaved caspase-3的表达降低,Cleaved caspase-12、CHOP阳性细胞比率减少,脊髓组织细胞凋亡比率减少(均P<0.01)。结论 EDA对脊髓损伤有保护作用,机制可能与其抑制SCI后脊髓细胞的ERS和脊髓细胞的凋亡有关。Objective To investigate the effects of edaravone （EDA） on cell apoptosis induced by endoplasmic reticu？lum stress （ESR） after spinal cord injury （SCI） in rats. Methods Thirty-six healthy adult SD rats were randomly divided in-to three groups （12 rats for each group）：Sham group, SCI group and EDA group. The rat model of SCI was made by Allen’s method and the sham group was only received laminectomy and kept the spinal cord intact. Rats in sham group and SCI group accepted the same volume and frequency of saline injection as EDA group. The EDA group was given 10 mg/kg EDA once every 12 h intraperitoneally. Three days after injuring, the spinal cords were harvested, and the protein levels of C/EBP homologous protein （CHOP）, Cleaved caspase-12 and Cleaved caspase-3 were detected by Western blot assay. Immunofluo？rescence staining was used to analyze the positive ratio of caspase-12 and CHOP in spinal cord of three groups. Meanwhile, TUNEL staining was used to identify cell apoptosis of spinal cord. Results Compared with sham group, the protein levels of CHOP, Cleaved caspase-12 and Cleaved caspase-3 were obviously higher in SCI group （P〈0.01）;the proportion of Cas？pase-12 and CHOP positive cells was significantly increased （P〈0.01）, and the apoptotic rates were also significantly in？creased in spinal cord （P〈0.01）. However, compared with SCI group, the protein levels of CHOP , Cleaved caspase-12 and Cleaved caspase-3 were significantly decreased in EDA group （P〈0.01）;the proportion of Caspase-12 and CHOP positive cells was significantly reduced （P〈0.01）, and the apoptotic rates were also significantly decreased in spinal cord （P〈0.01）. Conclusion EDA has neuroprotective potential to spinal cord injury. The mechanism of its neuroprotective effect may asso？ciate with its inhibitory effect to the cell apoptosis induced by endoplasmic reticulum stress after SCI.

关 键 词：脊髓损伤 细胞凋亡 半胱氨酸天冬氨酸蛋白酶3 半胱氨酸天冬氨酸蛋白酶12 模型 动物 大鼠 Sprague-Dawley 内质网应激 依达拉奉 转录因子CHOP 

分 类 号：R651.2[医药卫生—外科学]