白藜芦醇通过核因子E2相关因子2/抗氧化反应元件通路对脊髓损伤小鼠的抗氧化和抗抑郁作用

Antioxidant and antidepression effects of resveratrol on spinal cord injury mice by nuclear factor erythroid 2-related factor 2/ARE pathway

目的观察白藜芦醇(RES)对脊髓损伤(SCI)小鼠炎性反应和抑郁情绪的影响。方法采用改良的Allen’s击打法制作SCI小鼠模型,采用随机数字表将小鼠分为假手术组、对照组和治疗组。术后3 d,干湿称重法检测小鼠损伤侧脊髓含水量,二氢乙锭(HEt)染色检测活性氧(ROS)的积累,酶联免疫吸附试验(ELISA)试剂盒检测超氧化物歧化酶(SOD)、丙二醛(MDA)、白细胞介素(IL)-1β和肿瘤坏死因子-α(TNF-α)的表达。并通过核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)和SOD1蛋白表达探究其作用机制。术后28 d,我们通过悬尾试验、糖水偏好试验来评估白藜芦醇抗抑郁作用。组间比较采用t检验和单因素方差分析。结果治疗组损伤部位含水量(78.85±2.57)低于对照组(81.26±3.12),差异有统计学意义(n=5,t=-2.464,P<0.05)。治疗组小鼠糖水偏嗜度[(62.17±3.83)%]高于对照组[(55.63±3.69)%],差异有统计学意义(n=8,t=6.883,P<0.05)。SCI后28 d,治疗组小鼠不动时间[(205.80±5.03) s]低于对照组[(256.15±8.20) s],差异有统计学意义(n=8,t=-14.573,P<0.05)。治疗组ROS的积累[(2 523.23±89.22)%]低于对照组[(3 219.50±101.60)%],差异有统计学意义(n=5,t=-13.825,P<0.05)。治疗组SOD活力[(59.20±9.22) U/mg]高于对照组[(42.50±8.52) U/mg],差异有统计学意义(n=5,t=3.547,P<0.05)。治疗组减少MDA含量[(6.23±1.52) nmol/mg]高于对照组[(4.92±1.60) nmol/mg],差异有统计学意义,(n=5,t=2.361,P<0.05)。治疗组IL-1β[(45.32±11.23) ng/ml]低于对照组[(65.21±9.02) ng/ml],治疗组TNF-α[(735.32±33.29) ng/ml]低于对照组[(821.22±29.80) ng/ml],差异有统计学意义(n=5,t=-3.534,P<0.05;n=5,t=-3.944,P<0.05)。治疗组Nrf2(1.23±0.52)高于对照组(0.92±0.65),差异有统计学意义(n=5,t=2.302,P<0.05)。治疗组HO-1和SOD1(1.23±0.32、2.02±0.26)高于对照组(0.89±0.22、0.92±0.31),差异有统计学意义(n=5,t=3.306,P<0.05;n=5,t=8.731,P<0.05)。结论小鼠SCI后,白藜芦醇通过激活Nrf2通路对小鼠的抑郁情绪和炎性反应有明显改善作用。

Objective To investigate the influence of resveratrol on inflammation and depression in spinal cord injury mice.Methods Spinal cord injury(SCI)mouse model established by modified Allen′s hitting method.The mice were divided into sham group,control group and treatment group by random number table.At 3 days after SCI,the water content of the spinal cord was determined by dry and wet weighing method.The accumulation of reactive oxygen species(ROS)was determined by hematoxylin and eosin(HE)staining.Enzyme linked immunosorbent assay(ELISA)kit was used to detect the expression of superoxide dismutase(SOD),malondialdehyde(MDA),interleukin(IL)-1βand tumor necrosis factor-α(TNF-α).Western botting was used to detect the expression and the effect of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1)and SOD1.At 28 days after SCI,tail suspension and sucrose preference test was used to evaluate the anti-despondent effect of resveratrol.Comparison between groups was performed by t test and one-way ANOVA.Results The water content of injury site in the treatment group(78.85±2.57)was lower than that in the control group(81.26±3.12),the difference was statistically significant(n=5,t=-2.464,P<0.05).The degree of preference for sugar and water in the treatment group[(62.17±3.83)%]was higher than that in the control group[(55.63±3.69)%],the difference was statistically significant(n=8,t=6.883,P<0.05).28 days after SCI,the immobility time of mice in the treatment group[(205.80±5.03)s]was lower than that in the control group[(256.15±8.20)s],the difference was statistically significant(n=8,t=-14.573,P<0.05).The accumulation of ROS in the treatment group[(2523.23±89.22)%]was lower than that in the control group[(3219.50±101.60)%],the difference was statistically significant(n=5,t=-13.825,P<0.05).The activity of SOD in the treatment group[(59.20±9.22)U/mg]was significantly higher than that in the control group[(42.50±8.52)U/mg,n=5,t=3.547,P<0.05].The content of MDA in the treatment group decreased by(6.23±1.52)nmol/mg,which was significantly higher than that in the control group[(4.92±1.60)nmol/mg,n=5,t=2.361,P<0.05).Treatment group IL-1β[(45.32±11.23)ng/ml]was lower than that of the control group[(65.21±9.02)ng/ml]and the treatment group TNF-α[(735.32±33.29)ng/ml]was lower than that in the control group[(821.22±29.80)ng/ml],the difference was statistically significant(n=5,t=-3.534,P<0.05;n=5,t=-3.944,P<0.05).Nrf2 in the treatment group(1.23±0.52)was higher than that in the control group(0.92±0.65),the difference was statistically significant(n=5,t=2.302,P<0.05).HO-1 and SOD1 in the treatment group(1.23±0.32,2.02±0.26)was higher than that in the control group(0.89±0.22,0.92±0.31),the difference was statistically significant(n=5,t=3.306,P<0.05;n=5,t=8.731,P<0.05).Conclusion Resveratrol alleviated the depression and inflammation after spinal cord injury by Nrf2 pathway.