芒果苷在大鼠脊髓缺血再灌注性损伤中的保护作用及机制研究

Effects of mangiferin on spinal cord ischemia/reperfusion injury in rats

目的探索芒果苷(MAN)对大鼠脊髓缺血再灌注损伤的保护作用及相关分子机制。方法腹主动脉夹闭法制备大鼠脊髓缺血再灌注损伤模型,采用前瞻性分析,实验分3组:对照组、模型组和实验组。实验组利用50 mg/kg芒果苷干预治疗,对照组和模型组给予等量生理盐水腹腔注射。于造模后第7天和14天检测各组大鼠下肢运动功能;14 d利用ELISA法检测肝脏组织中丙二醛(MDA)含量、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)以及谷胱甘肽过氧化氢酶(GSH-px)活性;化学定量法及Western blotting法分别检测肝脏组织中一氧化氮(NO)和内皮型一氧化氮合酶(eNOS)、Bax、Bcl-2蛋白表达;14 d后取脊髓组织进行快蓝/HE(LFB/HE)染色和eNOS、Bax免疫组织化学染色。结果BBB评分显示,第7天,与对照组比较,模型组下肢运动功能评分明显下降,而实验组较模型组有升高(P<0.05);第14天,实验组BBB评分显著高于模型组(P<0.05);ELISA法显示,模型组大鼠脊髓组织MDA含量显著高于对照组,CAT、SOD和GSH-px显著高于对照组,而实验组MDA含量显著低于模型组,CAT、SOD和GSH-px显著高于模型组(P<0.05);化学定量法和Western blotting显示,模型组肝脏组织中NO和eNOS、Bax表达量显著高于对照组,而Bcl-2显著低于对照组(P<0.05),而实验组肝脏组织中NO和eNOS、Bax表达量显著低于模型组,而Bcl-2显著高于模型组(P<0.05)。免疫组织化学染色显示eNOS、Bax平均光密度实验组显著低于模型组(P<0.05)。结论芒果苷可通过抗氧化应激和细胞凋亡保护大鼠脊髓缺血再灌注性损伤。

Objective To explore the effects of mangiferin on the spinal cord ischemia/reperfusion injury in rats and its potential mechanism. Methods Spinal cord ischemia/reperfusion injury model was made by abdominal aortic clipping method. All rats were divided into 3 groups,the control group,the model group and the test group. Lower limb motor function was valued at the 7 th and 14 th day post surgery via BBB scores. At the 14 th day,malondialdehyde（ MDA）,catalase（ CAT）,superoxide dismutase（ SOD） and glutathione peroxidase（ GSH-px） were detected by elisa. Nitric oxide was detected by chemical quantitative method,and endothelial nitric oxide synthase（ eNOS）,Bax and Bcl-2 were valued by western blotting. Besides,eNOS and Bax were detected by immunohistochemical staining. Results At the 7 th day,compared with the control group,the scores of lower limb motor function decreased significantly in the model group,but the experimental group was higher than the model group（ P 〈 0. 05）. While at the 14 th day,BBB scores in the test group was markedly higher than the model group（ P 〈 0. 05）. Elisa showed that the contents of CAT,SOD and GSH-px in the spinal cord were significantly higher than those in the control group,while the contents of CAT,SOD and GSH-px were significantly higher in the experimental group than those in the control group（ P 〈 0. 05）. Western blotting and immunohistochemical staining showed that the expression of NO and eNOS and Bax in the liver tissue of model group were significantly higher than those of the control group,while Bcl-2 was significantly lower than that of the control group（ P 〈 0. 05）,while the expression of NO and eNOS and Bax in liver tissue of experimental group were significantly lower than those of model group,while Bcl-2 was significantly higher than that in the model group（ P 〈 0. 05）. Conclusion Mangiferin can significantly protect the spinal cord ischemia/reperfusion injury in rats via anti-oxidative stress and anti-apoptosis.