温肾醒脑方干预对血管性痴呆大鼠氧化应激的影响

Effect of Wenshen Xingnao decoction on oxidative stress in vascular dementia rats

目的观察温肾醒脑方对血管性痴呆(VD)大鼠氧化应激的影响及其机制。方法用随机数字表法将制模成功大鼠分为3组:模型组、中药组、西药组,每组15只;另取15只正常大鼠作为正常组。于术后第2天,正常组和模型组灌胃0.9%NaCl;中药组给予2 g·kg^-1·d^-1温肾醒脑方浓缩液10 mL,ig;西药组给予0.6 mg·kg^-1·d^-1吡拉西坦水溶液10 mL,ig,均1次/天,连续给药14 d。用酶联免疫吸附法分别检测血清和脑组织的丙二醛(MDA)、超氧化物歧化酶(SOD)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、乙酰胆碱酯酶(AchE)和胆碱乙酰转移酶(ChAT)指标,用免疫组织化学染色法检测大鼠脑组织海马区的核转录因子2(Nrf2)、血红素加氧酶-1(HO-1)和醌氧化还原酶-1(NQO-1)的蛋白表达。结果正常组、模型组、西药组和中药组的血清MDA(nmoL·mL^-1)分别为5.25±0.49,10.79±1.15,6.75±0.72和6.34±0.68;这4组的血清SOD(U·mL^-1)分别为97.63±15.62,40.39±7.17,70.66±13.27和76.01±13.89;这4组的血清IL-1β(pg·g-1)分别为2057.1±204.3,3566.7±227.5,2508.1±141.6和2353.9±125.8;这4组的Nrf2蛋白表达水平分别为1.48±0.50,0.39±0.32,0.97±0.42和1.16±0.45;这4组的HO-1蛋白表达水平分别为1.35±0.37,0.63±0.15,1.07±0.26和1.13±0.23;这4组的NQO-1蛋白表达水平分别为1.72±0.34,1.05±0.21,1.41±0.26和1.53±0.23。上述指标:模型组与正常组比较,差异均有统计学意义(均P<0.05);西药组和中药组与模型组比较,差异均有统计学意义(P<0.05)。基因结果的趋势与蛋白一致。结论温肾醒脑方通过激活Nrf2-抗氧化反应元件信号通路,抑制VD大鼠氧化应激及炎性反应损伤,发挥神经保护作用。

Objective To investigate the effect of Wenshen Xingnao decoction on oxidative stress in vascular dementia(VD)rats,and to explore its antioxidative mechanism.Methods Rats were divided into 4 groups:normal group,model group,western medicine group and traditional Chinese medicine group.VD rat model was constructed.Normal group and model group:normal saline,intragastric(ig);Chinese medicine group:Wenshen Xingnao Formula 10 mL,2 g·kg^-1·d^-1,ig;Western medicine group:Piracetam aqueous solution 10 mL,0.6 mg·kg^-1·d^-1,ig;1 time/d,14 d continuous administration.Enzyme linked immunosorbent assay was used to detect serum and brain tissues of malondialdehyde(MDA),superoxide dismutase(SOD),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),acetylcholinesterase(AchE),and choline acetyltransferase(ChAT).The protein expression of nuclear transcription factor 2(Nrf2),heme oxygenase-1(HO-1),and quinone oxidoreductase-1(NQO-1)in the hippocampus were determined by immunohistochemical staining.Results Serum MDA(nmoL·mL^-1)in normal group,model group,western medicine group,and traditional Chinese medicine group were respectively 5.25±0.49,10.79±1.15,6.75±0.72,6.34±0.68;serum SOD(U·mL^^-1)in the 4 groups were respectively 97.63±15.62,40.39±7.17,70.66±13.27,76.01±13.89;serum IL-1β(pg·g^-1)in the 4 groups were respectively 2057.1±204.3,3566.7±227.5,2508.1±141.6,2353.9±125.8;the level of Nrf2 protein in the 4 groups were respectively 1.48±0.50,0.39±0.32,0.97±0.42,1.16±0.45;the levels of HO-1 protein in the 4 groups were 1.35±0.37,0.63±0.15,1.07±0.26,1.13±0.23;the levels of NQO-1 protein in the 4 groups were 1.72±0.34,1.05±0.21,1.41±0.26,1.53±0.23.Compared between model group and normal group,the differences of the factors were significant(all P<0.05);compared between western medicine group,traditional Chinese medicine and model group,the differences of the factors were significant(all P<0.05).Gene results are consistent with protein trends.Conclusion Wenshen Xingnao decoction inhibits oxidative stress and inflammatory response injury in VD rats by activating Nrf2-antioxidant response element signaling pathway,and exerts neuroprotective effects.