通窍活血汤对蛛网膜下腔出血大鼠的神经保护作用及机制研究
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R743.35

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浙江省中医药科技计划项目(2021ZB285);嘉兴市科技计划民生科技创新研究项目(2021AD300222)


Neuroprotective Effect and Mechanism of Tongqiao Huoxue Decoction on Rats with Subarachnoid Hemorrhage
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    摘要:

    目的:观察通窍活血汤对蛛网膜下腔出血(SAH) 大鼠的神经保护作用,并探讨其作用机制。方 法:将60 只SD 大鼠随机分为假手术组,模型组,通窍活血汤低、中、高(3、6、12 g/kg) 剂量组各12 只。 采用改良二次枕大池注血法制作SAH 大鼠模型。造模成功后,通窍活血汤低、中、高剂量组大鼠分别灌胃对 应剂量的通窍活血汤溶液治疗,假手术组和模型组大鼠灌胃等量的0.9%氯化钠溶液,连续灌胃7 d。于治疗第 1、3、5、7 天评估各组大鼠Bederson 神经功能评分;每组各取6 只大鼠,在治疗第3、7 天分批取材,采用酶 联免疫吸附法检测各组大鼠海马组织肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β) 水平;免疫组织化 学法测定各组大鼠醌氧化还原酶1(NQO-1)、核因子E2 相关因子2(Nrf2) 蛋白表达。结果:与假手术组比 较,模型组大鼠在治疗第1、3、5、7 天的Bederson 神经功能评分均升高(P<0.05);通窍活血汤中、高剂量 组大鼠在治疗第5、7 天的Bederson 神经功能评分均较模型组降低(P<0.05)。与假手术组比较,模型组大鼠 治疗第3、7 天海马组织中TNF-α、IL-1β 含量明显升高(P<0.01),海马齿状回区NQO-1、Nrf2 蛋白表达明 显降低(P<0.01);与模型组比较,通窍活血汤低、中、高剂量组大鼠治疗第3、7 天海马组织中TNF-α、 IL-1β 含量降低(P<0.01),通窍活血汤低、中、高剂量组大鼠海马齿状回区NQO-1、Nrf2 蛋白表达升 高(P<0.05)。结论:通窍活血汤可通过抗氧化应激,减轻炎性反应来改善SAH 大鼠神经功能,其机制与调 节Nrf2-抗氧化反应元件(Nrf2-ARE) 信号通路下游NQO-1、Nrf2 蛋白水平有关。

    Abstract:

    Abstract: Objective: To observe the neuroprotective effect of Tongqiao Huoxue Decoction on rats with subarachnoid hemorrhage (SAH) and to discuss its mechanism. Methods:A total of 60 SD rats were randomly divided into the sham-operation group, the model group, the low-dose, medium-dose and high-dose of Tongqiao Huoxue Decoction group (3,6 and 12 g/kg),with 12 rats in each group. The SAH rat models were established by modified secondary blood injection into occipital cistern. After successful modeling, rats in the low- dose, medium- dose and high- dose of Tongqiao Huoxue Decoction groups were treated with Tongqiao Huoxue Decoction solution of corresponding dose respectively,and rats in the sham-operation group and the model group were treated with 0.9% sodium chloride solution of the same amount by gavage for consecutive day. The Bederson neurological function scores of rats in each group were evaluated on the first,third,fifth and seventh day of treatment;six rats in each group were taken in batches on the third and seventh day of treatment, and levels of tumor necrosis factor- α (TNF- α) and interleukin- 1β (IL- 1β) in the hippocampus of rats in each group were detected by enzyme- linked immunosorbent assay;the protein expressions of quinone oxidoreductase (NQO- 1) and nuclear factor erythroid 2- related factor 2 (Nrf2) were determined by immunohistochemistry. Results: Compared with those in the sham- operation group, the Bederson neurological function scores in the model group were higher on the first,third,fifth and seventh day of treatment (P<0.05);the Bederson neurological function scores of rats in the Tongqiao Huoxue Decoction group of of medium-dose and high-dose were decreased when compared with those in the model group on the fifth and seventh day of treatment (P<0.05). Compared with those in the sham-operation group,the levels of TNF-α and IL-1β in the hippocampus of rats in the model group were significantly increased on the third and seventh day of treatment (P<0.01), the protein expressions of NQO-1 and Nrf2 in the hippocampal dentate gyrus were significantly decreased (P<0.01);compared with those in the model group,the levels of TNF- α and IL-1β in the hippocampus of rats in the low- dose, medium- dose and high- dose of Tongqiao Huoxue Decoction group were decreased on the third and seventh day after treatment (P<0.01),and the protein expressions of NQO-1 and Nrf2 in the hippocampal dentate gyrus in the low- dose, medium- dose and high- dose of Tongqiao Huoxue Decoction group were increased (P<0.05). Conclusion:Tongqiao Huoxue Decoction can improve the neurological function of SAH rats by resisting oxidative stress and reducing the inflammatory reactions. Its mechanism is related to regulating the protein levels of NQO- 1 and Nrf2 downstream of the Nrf2 to antioxidant responsive element (Nrf2-ARE) signal pathway.

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魏绪旺,朱平,郭志丽,金敏艳,沈斌,孙长忠,刘鹏鹏,高晨琛.通窍活血汤对蛛网膜下腔出血大鼠的神经保护作用及机制研究[J].新中医,2023,55(7):15-20

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  • 在线发布日期: 2023-04-12
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