畲药异叶茴芹茎杆挥发油对LPS/D-GalN诱导的小鼠急性肝损伤保护作用研究

    Investigation on the Protective Effect of the Essential Oil from the Stem of Pimpinella Diversifolia on LPS/D-GalN-induced Acute Liver Injury in Mice

    • 摘要:
      目的 探究异叶茴芹茎杆挥发油(Pimpinella diversifolia stem essential oil,PDSEO)对脂多糖(lipopolysaccharide,LPS)/D-氨基半乳糖(D-galactosamine,D-GlaN)诱导小鼠急性肝损伤保护作用及其可能的分子机理。
      方法 同时蒸馏萃取法制备PDSEO,并采用气质联用色谱法(GC/MS)鉴定化学成分;以LPS/D-GalN为诱导剂制备急性肝损伤模型,检测血清ALT、AST和ALP水平,通过流式多因子检测(CBA)法分析血清中炎症因子水平; HE染色分析肝组织病变情况,肝组织匀浆检测MDA水平,荧光定量PCR分析肝组织炎症因子和抗氧化酶相关基因表达水平,TUNEL法分析肝细胞凋亡情况,综合评价PDSEO保肝活性和机制。
      结果 共鉴定PDSEO中46个成分,其中萜类化合物34种。PDSEO低(50 mg·kg−1)、高(100 mg·kg−1)剂量组均可显著降低LPS/D-GalN诱导的血清ALT、AST和ALP升高;也可有效降低肝组织炎症基因IL-6TNF-αMCP-1表达以及血清中炎症因子水平;PDSEO低、高剂量组处理后肝组织中SOD1SOD2CATGpx4等抗氧化相关基因表达上调,同时肝组织中SOD、CAT酶活力和GSH水平显著升高,而脂质过氧化产物MDA显著下降;HE和TUNEL凋亡染色进一步证实,PDSEO低、高剂量组均可有效抑制肝细胞凋亡,减轻肝组织损伤。
      结论 异叶茴芹茎杆富含挥发油,其可通过抑制肝组织过度炎症反应,提高抗氧化系统水平降低肝组织过度氧化应激,减少肝细胞凋亡,有效缓解LPS/D-GalN诱导的小鼠急性肝损伤,在预防肝组织损伤药物开发方面表现出潜在开发利用价值。

       

      Abstract:
      OBJECTIVE To explore the protective effect of the Pimpinella diversifolia stem essential oil(PDSEO) on lipopolysaccharide(LPS)/D-galactosamine(D-GalN) induced acute liver injury in mice and its possible molecular mechanism.
      METHODS PDSEO was prepared through the simultaneous distillation and extraction method and its chemical composition were identified by GC/MS. The acute liver injury model was established using LPS/D-GalN as the inducer. The serum levels of ALT, AST, and ALP were determined, and the serum levels of inflammatory factors were analyzed by cytometric bead array assay(CBA). The pathological changes of liver tissues were analyzed by HE staining. The level of malondialdehyde(MDA) was detected using liver tissue homogenates. The expression levels of genes related to inflammatory factors and antioxidant enzymes in liver tissues were analyzed by fluorescence quantitative PCR. The apoptosis of hepatocytes was analyzed by the TUNEL method. The hepatoprotective activity and mechanism of PDSEO were comprehensively evaluated.
      RESULTS A total of 46 components of PDSEO were identified, among which 34 were terpenoids. Both low(50 mg·kg−1) and high(100 mg·kg−1) doses of PDSEO could conspicuously reduce the elevation of serum ALT, AST and ALP induced by LPS/D-GalN. It could also efficaciously decrease the expression of inflammatory genes IL-6, TNF-α and MCP-1 in liver tissue as well as the level of serum inflammatory factors. After the treatment with low and high doses of PDSEO, the expression of anti-oxidation-related genes such as SOD1, SOD2, CAT and Gpx4 in liver tissue was up-regulated. Meanwhile, the activities of SOD and CAT enzymes and the GSH levels in liver tissue were significantly augmented, while the lipid peroxidation product MDA was significantly diminished. HE and TUNEL apoptotic staining of liver tissue sections further confirmed that both the low and high dose PDSEO groups could effectively alleviate hepatocyte apoptosis and liver tissue injury.
      CONCLUSION The stems of Pimpinella diversifolia abound in essential oil, which is capable of suppressing excessive inflammation of liver tissue, enhancing the level of the antioxidant system, alleviating the emergency response of excessive oxidation of liver tissue, decrease liver cell apoptosis, effectively mitigating LPS/D-GalN-induced acute liver injury in mice, and demonstrating potential development and utilization value in the development of drugs for preventing liver tissue injury.

       

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