黑骨藤对脊髓损伤大鼠神经功能恢复的影响及其机制研究

Effect of Periploca forrestii Schltr on nerve function recovery in rats with spinal cord injury and its mechanism

目的:研究黑骨藤对大鼠脊髓损伤后神经功能恢复的影响及其机制。方法:将30只成年SD大鼠随机分为假手术组、脊髓损伤组和黑骨藤组。假手术组仅切除椎板,脊髓损伤组建立钳夹型大鼠脊髓损伤模型,黑骨藤组建立钳夹型大鼠脊髓损伤模型后给予20 mg/kg黑骨藤乙醇提取物。术后采用Basso-Beattie-Bresnahan(BBB)量表进行运动功能评分,并通过斜板试验最大角度来评估脊髓功能恢复情况,苏木精—伊红(HE)染色观察脊髓组织病理形态变化,酶联免疫吸附试验(ELISA)法检测损伤段脊髓组织超氧化物歧化酶(SOD)、诱导性一氧化氮合成酶(iNOS)、丙二醛(MDA)及一氧化氮(NO)含量。Western blotting法检测环氧合酶-2(COX-2)和白介素1β(IL-1β)蛋白表达。结果:脊髓损伤组和黑骨藤组术后1 d、3 d、7 d、21 d斜板试验最大角度和BBB评分均明显低于假手术组,脊髓损伤组术后7 d、21 d斜板试验最大角度和BBB评分均明显低于黑骨藤组(均P<0.05)。与脊髓损伤组比较,黑骨藤组脊髓组织iNOS、MDA、NO水平及COX-2、IL-1β蛋白表达量显著升高,SOD活性明显降低(P<0.05),脊髓组织病理损伤明显减轻。结论:黑骨藤可能通过抑制氧化应激反应和炎症反应促进脊髓损伤大鼠神经功能恢复。

Objective:To investigate the effect of Periploca forrestii Schltr(PFS)on the recovery of neurological function in rats with spinal cord injury(SCI)and its possible mechanism.Methods:Thirty SD rats were randomly divided into sham-operated group,SCI group,and PFS group.The SCI model was established by vertical clamp.The sham-operated group rats were given laminectomy only,and rats in PFS group were given 10 mL/kgethanolic extracts of PFS after modeling.Basso-Beattie-Bresnahan(BBB)scalewas used to evaluate the motor function of rats after operation.Maximum angle of inclined plate test was used to evaluate the recovery of spinal cord function.Hematoxylin-eosin(HE)staining was employed to observe the pathological changes of spinal cord tissues.The activities of superoxide dismutase(SOD)and inducible nitric oxide synthase(iNOS)and the contents of malondialdehyde(MDA)and nitrogen monoxide(NO)in the injured spinal cord tissue were detected by enzyme-linked immunosorbent assay(ELISA).The protein expressions of cyclooxygenase-2(COX-2)and interleukin-1β(IL-1β)were detected by western blotting.Results:The BBB score and the maximum angle of inclined plate test in SCI and PFS groupsweredecreasedafter 1 d,3 d,7 d,and 21 d of operation compared to sham-operated group,and the BBB score and the maximum angle of inclined plate test on day 7 and 21 post-operation were significantly higher in PFS group compared to SCI group(P<0.05).Compared with SCI group,the iNOS,MDA and NO contents as well as the protein expressions of COX-2 and IL-1βin spinal cord tissues were increased,while the SOD activity wasdecreased in PFS group(P<0.05).The pathological injury of spinal cord tissues was relieved obviously in PFS group.Conclusion:PFS may promote the recovery of neurological function in SCI rats by alleviating oxidative stress and inflammatory response.