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    基于D-半乳糖诱导的氧化应激小鼠模型的丹参-红花-三七配伍抗氧化应激损伤活性评价及机制研究

    Study on the Anti-Oxidant Stress Injury Activity of Salviae Miltiorrhizae Radix Et Rhizoma-Carthami Flos-Notoginseng Radix et Rhizoma Combination based on the D-galactose Induced Oxidative Stress Mice Model

      摘要
    • 摘要:
      目的  基于D-半乳糖诱导的氧化应激小鼠模型研究丹参-红花-三七配伍的抗氧化应激损伤活性。
      方法 采用UPLC-QTOF-MS鉴定丹参-红花-三七配伍样品的化学组成；采用腹腔注射D-半乳糖建立氧化应激小鼠模型，评价丹参、红花、三七单用和配伍对小鼠氧化应激损伤的影响；采用微量法检测血清氧化应激标志物超氧化物歧化酶(superoxide dismutase，SOD)、丙二醛(malondialdehyde，MDA)、谷胱甘肽过氧化物酶(glutathioneperoxidase，GSH-Px)和肝损伤标志物谷丙转氨酶(aminotransferase，ALT)、谷草转氨酶(aminotransferase，AST)的含量；采用ELISA检测肝组织中炎症因子IL-1β、IL-6和TNF-α的含量；Real-time PCR检测肝组织中Casp3、Bax、Bcl-2、IκK、IκB和NF-κB mRNA的表达水平。
      结果 从丹参、红花、三七和配伍样品中分别鉴定出17、11、40和68个化合物；丹参、红花、三七单用和配伍对小鼠体质量和脏器指数(心脏、肝脏、脾脏、肺、肾脏和胰腺)无显著影响；与对照组相比，模型组小鼠血清中SOD和GSH-Px的含量显著降低，MDA和ALT的含量显著升高，肝组织中IL-1β、IL-6、TNF-α蛋白和Casp3、IκK、IκB、NF-κB mRNA的表达水平显著升高。与模型组相比，红花和三七显著升高血清SOD的含量，配伍组血清中SOD和GSH-Px的含量显著升高且MDA含量显著降低；三七、丹参和配伍组血清ALT含量显著降低；红花、三七、丹参和配伍组肝组织Casp3 mRNA的表达水平显著降低，且红花和配伍组肝组织中Bcl-2/Bax mRNA显著升高；三七、丹参和配伍组肝组织IL-1β、IL-6、TNF-α蛋白和IκK、IκB、NF-κB mRNA的表达水平显著降低；与丹参、红花或三七组相比，配伍组显著增加血清SOD含量和肝组织Bcl-2/Bax mRNA表达水平，并显著降低血清MDA和ALT含量、肝组织IL-1β、IL-6、TNF-α蛋白和IκK、IκB、NF-κB mRNA的表达水平。
      结论 丹参-红花-三七配伍可协同改善D-半乳糖诱导的发氧化应激，其机制可能与激活酶氧化系统、减轻氧化应激损伤和抑制炎症通路激活有关。

       

      Abstract:
      OBJECTIVE  To investigate the anti-oxidative stress injury effects of the Salviae Miltiorrhizae Radix Et Rhizoma-Carthami Flos-Notoginseng Radix et Rhizoma combination based on the D-galactose induced oxidative stress mice model.
      METHODS  The chemical compositional of the Salviae Miltiorrhizae Radix Et Rhizoma-Carthami Flos-Notoginseng Radix et Rhizoma compatible sample was identified by UPLC-QTOF-MS. A mice model of oxidative stress was established by intrabitoneal injection of D-galactose, and the effects of Salviae Miltiorrhizae Radix Et Rhizoma, Carthami Flos, Notoginseng Radix et Rhizoma on oxidative stress injury were evaluated. The contents of superoxide dismutase(SOD), malondialdehyde(MDA), glutathione peroxidase(GSH-Px) in serum and alanine aminotransferase(ALT), oxalic aminotransferase(AST) in liver tissues were detected by micromethod. The levels of inflammatory factors IL-1β, IL-6 and TNF-α in liver tissues were detected by ELISA. The mRNA expression levels of Casp3, Bax, Bcl-2, IκK, IκB and NF-κB in liver tissues were detected by Real-time PCR.
      RESULTS  A total of 17, 11, 40, and 68 compounds were identified from Salviae Miltiorrhizae Radix Et Rhizoma, Carthami Flos, Notoginseng Radix et Rhizoma, and compatible sample, respectively. Body weight and organ index(heart, liver, spleen, lung, kidney and pancreas) were not significantly changed in each group. Compared with the Con group, the contents of SOD and GSH-Px were significantly decreased and the contents of MDA and ALT were significantly increased in serum of model group, and the expression levels of IL-1β, IL-6, TNF-α protein and Casp3, IκK, IκB, NF-κB mRNA in liver tissues of model group were significantly increased. Compared with the model group, Carthami Flos and Notoginseng Radix et Rhizoma significantly increased the content of SOD in serum, and the content of SOD and GSH-Px were significantly increased and the content of MDA was significantly decreased in serum of combination group; Notoginseng Radix et Rhizoma, Salviae Miltiorrhizae Radix Et Rhizoma and compatible sample significantly decreased serum ALT content; The expression level of Casp3 mRNA in liver tissues of all groups were significantly decreased, and the expression level of Bcl-2/Bax mRNA in liver tissues of Carthami Flos and combination groups were significantly increased; The expression levels of IL-1β, IL-6, TNF-α protein and mRNA of IκK, IκB and NF-κB in liver tissues of Notoginseng Radix et Rhizoma, Salviae Miltiorrhizae Radix Et Rhizoma and combination groups were significantly decreased. Compared with Salviae Miltiorrhizae Radix Et Rhizoma, Carthami Flos or Notoginseng Radix et Rhizoma groups, the SOD content in serum and Bcl-2/Bax mRNA expression level in liver tissues were significantly increased, and the contents of MDA and ALT in serum and the expression levels of IL-1β, IL-6, TNF-α protein and IκK, IκB, NF-κB mRNA in liver tissue were significantly decreased in combination groups.
      CONCLUSION  Salviae Miltiorrhizae Radix Et Rhizoma-Carthami Flos-Notoginseng Radix et Rhizoma combination can synergically improve the oxidative stress induced by D-galactose, and its mechanism may be related to activating the enzyme oxidation system, alleviating oxidative stress injury and inhibiting the activation of inflammatory pathways.

       

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