白藜芦醇通过AMPK/PGC-1α信号通路缓解三硝基丙酸诱导的亨廷顿病模型小鼠的作用研究

Effect of resveratrol on Huntington's disease modeled mice induced by 3-nitropropionic acid by targeting AMPK/PGC-1αsignaling pathway

目的以三硝基丙酸诱导亨廷顿病模型为切入点,从记忆功能、平衡协调能力、脑组织的生化指标以及腺苷酸活化蛋白激酶(AMPK)和过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)表达等多角度探讨白藜芦醇缓解亨廷顿病的作用机制。方法50只雄性3月龄无特定病原体级ICR小鼠,体重18~22g。按照随机数字表法将其分为正常对照组、模型组和白藜芦醇低、中、高剂量组,每组10只。除正常对照组外,其余各组均采用三硝基丙酸诱导亨廷顿病模型,模型组与正常对照组给予等量蒸馏水,白藜芦醇低、中、高剂量组分别给予50、100、200mg/kg白藜芦醇灌胃,1次/d。连续灌胃14d后检测各组小鼠记忆功能、平衡协调能力、脑组织中谷胱甘肽过氧化物酶(GPx),超氧化物歧化酶(SOD)、Na^(+)-K^(+)-ATPase酶表达水平、AMPK和PGC-1α阳性表达情况及脑组织中AMPK和PGC-1α蛋白表达量。结果与正常对照组比较,模型组小鼠的逃避潜伏期的延长,差异有统计学意义(P<0.05);与模型组比较,随白藜芦醇剂量的增加,白藜芦醇组小鼠的逃避潜伏期缩短,差异有统计学意义(P<0.05);空间搜索试验结果显示,模型组小鼠在第一象限与第三象限的出现频率基本相同(P>0.05),而正常对照组、白藜芦醇低、中和高剂量组小鼠第三象限显著低于第一象限,差异均有统计学意义(P<0.05)。与正常对照组比较,模型组小鼠的掉落潜伏期明显缩短,差异有统计学意义(P<0.05);与模型组比较,白藜芦醇低、中、高剂量组小鼠的掉落潜伏期均明显延长,差异均有统计学意义(P<0.05)。与正常对照组比较,模型组小鼠脑组织中GPx、SOD和Na^(+)-K^(+)-ATP酶的活力显著降低,差异均有统计学意义(P<0.05);与模型组比较,白藜芦醇低、中、高剂量组小鼠的脑组织中GPx、SOD和Na^(+)-K^(+)-ATPase酶的活力显著升高,差异均有统计学意义(P<0.05)。与正常对照组比较,模型组脑组织中AMPK和PGC-1α阳性表达和蛋白表达显著降低,差异均有统计学意义(P<0.05);与模型组比较,白藜芦醇各剂量组中AMPK和PGC-1α阳性表达和蛋白表达显著增加,差异均有统计学意义(P<0.05)。以上作用呈剂量依赖性。结论白藜芦醇可能通过改变AMPK/PGC-1α信号通路蛋白,进而调节小鼠脑组织中SOD、GPx和Na^(+)-K^(+)-ATPase酶活性,有效地改善亨廷顿病的发生和发展。

Objective Taking 3-nitropropionic acid induced Huntington's disease model as a breakthrough point,to investigate the mechanism of resveratrol in relieving Huntington's disease from multiple perspectives including memory function,balance coordination ability,biochemical indicators of brain tissue,and expression of adenylate activated protein kinase(AMPK)and peroxlsome proliferator-activated receptor-γcoactlvator-1α(PGC-1α),etc.Methods Fifty male 3-month-old SPF grade ICR mice(weighing 18-22 g)were randomly divided into five groups,namely normal control group,model group and resveratrol groups(low,middle and high dose groups),each group with 10 mice.Except for the normal control group,the other groups were modelled on Huntington's disease with trinitropropionic acid.The normal control group was given the same amount of distilled water,and the resveratrol low,medium,and high doses groups were given 50,100,and 200 mg/kg of resveratrol by gavage,1 day/time.After 14 days of continous gavage,memory function,balance and coordination ability,expression levels of GPx,SOD,Na^(+)-K^(+)-ATP ase in brain tissue,positive expression of AMPK and PGC-1α,and protein expression levels of AMPK and PGC-1αin brain tissue were detected.Results Compared with the normal control group,the escape latency of the model group mice was prolonged,and the difference was statistically significant(P<0.05);compared with the model group,as the dose of resveratrol increases,the escape latency of mice in the resveratrol group was shortened,and the difference was statistically significant(P<0.05).The results of the spatial search experiment showed that the frequency of appearance of mice in the first quadrant and the third quadrant of the model group was basically the same(P>0.05),while the third quadrant of the normal control group,low,medium and high doses of resveratrol groups was significantly lower than the first quadrant,the difference was statistically significant(P<0.05).Compared with the normal control group,the drop latency of mice in the model group was significantly shorter,and the difference was statistically significant(P<0.05);compared with the model group,the drop latency of mice in the low,medium,and high dose of resveratrol groups were significantly prolonged,and the differences were statistically significant(P<0.05).Compared with the normal control group,the activity of GPx,SOD and Na^(+)-K^(+)-ATPase in the brain tissue of the model group was significantly reduced,and the difference was statistically significant(P<0.05);compared with the model group,the activities of GPx,SOD and Na^(+)-K^(+)-ATPase of the brain tissues in the low,middle and high dose of resveratrol groups were significantly increased,and the differences were statistically significant(P<0.05).Compared with the normal control group,the positive expression and protein expression of AMPK and PGC-1αin the brain tissue of the model group were significantly reduced,and the differences were statistically significant(P<0.05);compared with the model group,the positive expression and protein expression of AMPK and PGC-1αin the brain tissue of low,middle and high dose of resveratrol groups were significantly increased,and the differences were statistically significant(P<0.05).The above effects were dose-dependent.Conclusion Resveratrol may be able to effectively ameliorate the odurrence and development of Huntington's disease by targeting the AMPK/PGC-1αsignaling pathway,thereby regulating SOD,GPx and Na^(+)-K^(+)-ATPase activity in the mice's brain tissues.