牛膝-白芍对肝阳上亢型帕金森病小鼠脑组织氧化应激的影响

Effects of Achyranthis bidentatae-Cynanchum otophyllum on oxidative stress of cerebral tissue in Parkinson’s disease mice with syndrome of hyperactivity of liver-Yang

目的探讨镇肝熄风汤的核心药对牛膝-白芍对肝阳上亢型帕金森病(PD)小鼠脑组织氧化应激的影响。方法将C57BL/6小鼠随机分为正常对照组、模型组、镇肝熄风汤组、牛膝组、白芍组、牛膝-白芍药对组,每组10只。采用附子煎液(4 g/kg)灌胃联合1-甲基-4苯基-1,2,3,6-四氢吡啶(25 mg/kg)腹腔注射方法复制肝阳上亢型PD小鼠模型。观察小鼠行为学变化和脑黑质神经元超微结构,检测小鼠脑黑质区酪氨酸羟化酶(TH)阳性神经元表达水平和脑黑质中总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、丙二醛(MDA)表达水平以及核因子E2相关因子2(Nrf2)、硫氧还蛋白还原酶1(Trxr1)、硫氧还蛋白互作蛋白(Txnip)mRNA和蛋白表达水平。结果与模型组比较,各给药组小鼠PD样行为和脑黑质神经元超微结构均有改善,TH阳性神经元表达水平,T-AOC(除白芍组外)、SOD(除白芍组外)表达水平以及Nrf2、Trxr1 mRNA相对表达量和蛋白表达水平均显著升高,MDA(除白芍组外)表达水平以及Txnip mRNA相对表达量和蛋白表达水平均显著降低(P<0.05)。牛膝-白芍药对组的干预效果优于牛膝组和白芍组(P<0.05),与镇肝熄风汤组作用相当(P>0.05)。结论牛膝-白芍药对可能以Nrf2为靶点抑制肝阳上亢型PD小鼠脑组织氧化应激水平,对脑部神经元起到保护作用,效果优于牛膝、白芍单味药材,能够发挥与组方相当的补肾柔肝、潜阳熄风的功效。

OBJECTIVE To explore the effects of Achyranthis bidentatae-Cynanchum otophyllum as core couplet medicinals of Zhengan xifeng decoction on oxidative stress of cerebral tissue in Parkinson’s disease(PD)mice with syndrome of hyperactivity of liver-Yang.METHODS C57BL/6 mice were randomly divided into normal control group,model group,Zhengan xifeng decoction group,A.bidntatae group,C.otophyllum group and couplet medicinals of A.bidentatae-C.otophyllum group,with 10 mice in each group.PD model of hyperactivity of liver-Yang was established by intragastric administration of Aconitum carmichaelii decoction(4 g/kg)and intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(25 mg/kg).The behavioral changes and ultrastructure of substantia nigra neurons in mice were observed.The expressions of tyrosine hydroxylase(TH)positive neurons in substantia nigra were detected.The expressions of total antioxidant capacity(T-AOC),superoxide dismutase(SOD)and malondialdehyde(MDA)in substantia nigra as well as mRNA and protein expressions of nuclear factor-erythroid 2-related factor 2(Nrf2),thioredoxin reductase 1(Trxr1),thioredoxin interacting protein(Txnip)were also determined.RESULTS Compared with model group,PD behavior and ultrastructure of substantia nigra neurons were all improved in administration groups.The expressions of TH positive neurons,T-AOC(except for C.otophyllum group)and SOD(except for C.otophyllum group),mRNA relative expression and protein expression levels of Nrf2 and Trxr1 were all increased significantly;the expression of MDA(except for C.otophyllum group)and mRNA relative expression and protein expression levels of Txnip were all decreased significantly(P<0.05).Intervention effect of couplet medicinals of A.bidentatae-C.otophyllum group was better than that of A.bidntatae group and C.otophyllum group(P<0.05),and the effect was similar to that of Zhengan xifeng decoction group(P>0.05).CONCLUSIONS The couplet medicinals of A.bidentatae-C.otophyllum can inhibit the level of oxidative stress in the cerebral tissue of PD mice with hyperactivity of liver-Yang by targeting Nrf2,and play a protective role on the brain neurons.Its effect is better than that of A.bidntatae and C.otophyllum,and it plays the same role as that of the formula in tonifying the kidney,softening the liver and suppressing the Yang.