莱菔硫烷对小鼠脊髓损伤后后肢功能的作用  

Effect of sulforaphane on hindlimb function after spinal cord injury in mice

作　　者：廖小俊[1] 袁继超[1] 朱海涛[1] 刘伟[1] 陈亚星[1] 胡胜利[1] 李兰[1] 林江凯[1] 

机构地区：[1]第三军医大学西南医院神经外科全军神经外科研究所全军神经创伤防治重点实验室,重庆400038

出　　处：《第三军医大学学报》2016年第10期1066-1072,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81401012);国家自然科学基金面上项目(81571214;81471261)~~

摘　　要：目的观察莱菔硫烷(sulforaphane,SFN)对小鼠脊髓损伤后炎症反应、胶质瘢痕、轴突再生的干预作用,探讨其在脊髓损伤中的作用机制。方法雌性ICR小鼠66只,按随机数字表法分为4组:假手术组(n=12)、单纯损伤组(n=18)、莱菔硫烷组[5 mg/(kg·d)SFN,ip,n=18]、溶剂对照组(等体积玉米油腹腔注射,n=18)。假手术组仅咬除椎板,余各组行脊髓夹伤术,制备脊髓夹伤模型。建模后1、7、14、21、28 d进行小鼠后肢运动功能评分(BMS评分);Western blot检测建模后7 d各损伤组小鼠脊髓GFAP、CSPG、P65表达情况,28 d后各损伤组GAP-43表达情况;ELISA检测建模后7 d各组小鼠脊髓TNF-α、IL-1β表达情况;免疫荧光染色观察建模后7 d各损伤组小鼠脊髓GFAP、CSPG、P65表达情况。结果假手术组BMS评分9分,各脊髓损伤组术后后肢运动功能均有不同程度恢复,术后28 d时单纯损伤组评分为(2.17±0.41)分,溶剂对照组(2.33±0.52)分,莱菔硫烷组(3.33±0.52)分,后者与前二者比较差异有统计学意义(P<0.05)。术后7 d行免疫荧光染色可见各脊髓损伤组星形胶质细胞活化,GFAP、CSPG、P65表达增加,单纯损伤组与溶剂对照组较莱菔硫烷组更明显。术后7 d莱菔硫烷组GFAP、CSPG、P65蛋白相对表达量均较单纯损伤组与溶剂对照组低(P<0.05),术后28 d莱菔硫烷组GAP-43蛋白相对表达量较单纯损伤组与溶剂对照组明显增加(P<0.05)。建模后7 d,莱菔硫烷组小鼠脊髓TNF-α、IL-1β表达较其余损伤组明显降低(P<0.05)。结论莱菔硫烷可减轻小鼠脊髓损伤后炎症反应,抑制胶质瘢痕形成,促进轴突再生,改善后肢运动功能。Objective To observe the effect of sulforaphane on the inflammatory reaction,glial scar and axon regeneration after spinal cord injury（ SCI） in mice,and investigate the role of sulforaphane after SCI. Methods Sixty-six female ICR mice were randomly divided into 4 groups,that is,sham-operation group（ group A,n = 12）,simple injury group（ group B,n = 18）,solvent control group（ group C,n = 18）,and sulforaphane treatment group（ group D,n = 18）. The mice in group A received laminectomy only,while the spinal cord of T11 segment of the mice in groups B,C and D was crushed to 0. 2 mm for 15 s with a vessel clamp. The mice in group D were intraperitoneally given 5 mg / kg sulforaphane immediately after SCI,while the ones in group C were given equivalent amount of corn oil. The Basso mouse score（ BMS） locomotor rating scale was used to evaluate behavioral consequences in 1,7,14,21,and 28 d after injury. Then the mice were sacrificed and the spinal cord tissue samples were harvested to observe the expression of P65,glial fibrillary acidic protein（ GFAP） and chondroitin sulfate proteoglycan（ CSPG） by immunofluorescence assay in7 d after injury. The protein expression of P65,GFAP and CSPG were determined by Western blotting in 7 d after injury and GAP-43 was detected in 28 d after injury. The protein expression of TNF-α and IL-1β were determined by ELISA in 7 d after injury. Results The BMS score in group A was 9. After SCI,hindlimb motor function in groups B,C and D showed varied degrees of recovery,and after 28 d,the BMS scores were2. 17 ± 0. 41 in group B,2. 33 ± 0. 52 in group C,and 3. 33 ± 0. 52 in group D. Group D was improved more remarkably than group B and group C（ P〈 0. 05）,but there was no significant difference between group B and group C（ P〈 0. 05）. In 7 d after SCI,astrocyte activation was observed in each SCI group by immunofluorescence assay,and the expression levels of P65,GFAP and CSPG were increased more in group B and group C than in group D. Western blotting

关 键 词：莱菔硫烷 脊髓损伤 胶质瘢痕 轴突再生 

分 类 号：R282.71[医药卫生—中药学] R364.3[医药卫生—中医学]