717解毒合剂调控VEGF/ERK通路对蝮蛇咬伤大鼠局部组织损伤的修复作用

    Repair Effect of 717 Jiedu Decoction on Local Tissue Damage Induced by Agkistrodon Halys Snakebite in Rats via Regulation of the VEGF/ERK Pathway

    • 摘要:
      目的  探讨717解毒合剂通过调控VEGF/ERK信号通路促进蝮蛇咬伤大鼠局部损伤组织修复的机制。
      方法 选取SPF级健康SD大鼠60只,随机分为正常对照组(对照组)、蝮蛇咬伤模型组(模型组)、抗蝮蛇毒血清组(血清组)和717解毒合剂高、中、低剂量组(717-H组、717-M组、717-L组)。通过苏木素-伊红(hematoxylin-eosin staining,HE)和马松(Masson)染色观察腓肠肌病理变化,免疫组化观察中性粒细胞趋化因子-1(cytokine-induced neutrophil chemoattractant-1,CINC-1)和细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)的表达,酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测血清中血管内皮生长因子(vascular endothelial growth factor,VEGF)、细胞外调节蛋白激酶(extracellular signal-regulated kinase,ERK)、神经生长因子(nerve growth factor,NGF)及干扰素-γ(interferon-γ,IFN-γ)的含量,Western blotting及逆转录实时荧光定量聚合酶链式反应(reverse transcription-quantitative polymerase chain reaction,RT-qPCR)分别检测创面组织中VEGF、ERK、NGF、IFN-γ的蛋白和mRNA表达水平,生化试剂盒检测活性氧(reactive oxygen species,ROS)水平。
      结果 模型组腓肠肌组织损伤明显,血清组和717解毒合剂各剂量组均有不同程度改善,尤以717-H组接近正常对照组。模型组胶原纤维沉积增加,ROS水平显著升高;抗蝮蛇毒血清和717解毒合剂可明显降低胶原沉积和ROS水平。模型组CINC-1和ICAM-1表达明显上升,抗蝮蛇毒血清和717解毒合剂可明显下调CINC-1和ICAM-1表达。VEGF、NGF、ERK和IFN-γ在模型组上调,血清组和717-H组表现出最佳抑制效果。
      结论 717解毒合剂可有效减轻组织纤维化、抑制氧化应激和炎症反应,其机制可能与其抑制VEGF/ERK信号通路有关,该研究为蝮蛇毒诱导的局部组织损伤防治提供新靶点。

       

      Abstract:
      OBJECTIVE To explore the mechanism by which 717 Jiedu Decoction promotes the repair of local damaged tissue in rats bitten by Agkistrodon halys through regulating the VEGF/ERK signaling pathway.
      METHODS  Sixty SPF-grade healthy SD rats were selected and randomly divided into the normal control group(control group), the Agkistrodon halys bite model group(model group), the anti-Agkistrodon halys venom serum group(serum group), and the high-, medium-, and low-dose group of 717 Jiedu Decoction(717-H group, 717-M group, 717-L group). Histopathological changes in the gastrocnemius muscle were observed using hematoxylin-eosin(HE) and Masson staining. Immunohistochemistry was used to detect the expression of cytokine-induced neutrophil chemoattractant-1(CINC-1) and intercellular adhesion molecule-1(ICAM-1). The levels of vascular endothelial growth factor(VEGF), nerve growth factor(NGF), extracellular signal-regulated kinase(ERK), and interferon-γ(IFN-γ) in serum were measured by enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expression levels of VEGF, ERK, NGF, and IFN-γ in wound tissue were detected by Western blotting and reverse transcription-quantitative polymerase chain reaction(RT-qPCR), respectively. Reactive oxygen species (ROS) levels were measured using a biochemical assay kit.
      RESULTS The model group showed marked tissue damage, while the serum group and each dose groups of 717 Jiedu Decoction exhibited varying degrees of improvement, with the 717-H group closely resembling the control group. The model group also showed increased collagen deposition and ROS levels, which were significantly reduced by anti-Agkistrodon halys venom serum and 717 Jiedu Decoction. CINC-1 and ICAM-1 expression was significantly upregulated in the model group, whereas their expression was downregulated by anti-Agkistrodon halys venom serum and 717 Jiedu Decoction. VEGF, NGF, ERK, and IFN-γ were upregulated in the model group, and both serum group and 717-H group showed strong inhibitory effects on these factors.
      CONCLUSION 717 Jiedu Decoction effectively reduces tissue fibrosis, oxidative stress, and inflammatory responses. Its mechanism may be related to its inhibition of the VEGF/ERK signaling pathway. The research provides a new target for the prevention and treatment of local tissue damage induced by Agkistrodon halys venom.

       

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