717解毒合剂调控VEGF/ERK通路促进蝮蛇咬伤大鼠局部组织损伤修复作用

717 JieDu Decoction regulates VEGF/ERK pathway and promotes local tissue damage repair in Agkistrodon halys bite rats

关键词:  717解毒合剂  蝮蛇咬伤  VEGF/ERK  局部组织损伤

Key words:  717 JieDu Decoction  agkistrodon snakebite  VEGF/ERK  local tissue injury

摘要:

摘要：目的 探讨717解毒合剂通过调控VEGF/ERK信号转导通路促进蝮蛇咬伤大鼠局部损伤组织修复机制。方法 选取SPF级健康SD大鼠60只，随机分为正常对照组、蝮蛇咬伤模型组、抗蝮蛇毒血清组和717解毒合剂高、中、低剂量组。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)?、Western Blot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)检测创面组织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)水平，生化试剂盒检测活性氧(reactive oxygen species, ROS)水平。 结果 蝮蛇咬伤模型组腓肠肌组织损伤明显，抗蝮蛇毒血清组和717解毒合剂各剂量组均有不同程度改善，尤以717解毒合剂高剂量组接近正常对照组。蝮蛇咬伤模型组胶原纤维沉积增加，ROS水平显著升高；717解毒合剂高剂量组胶原沉积和ROS水平均显著降低。717解毒合剂各剂量组CINC-1和ICAM-1表达下调，尤其是717解毒合剂高剂量组。VEGF、NGF、ERK和IFN-γ在蝮蛇咬伤模型组上调，抗蝮蛇毒血清组和717解毒合剂高剂量组表现出最佳抑制效果。结论 717解毒合剂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，研究为蝮蛇毒诱导的局部组织损伤防治提供新靶点。

Abstract:

ABSTRACT: OBJECTIVE To explore the mechanism by which 717 Jiedu Decoction promotes the repair of local damaged tissues in rats bitten by Agkistrodon halys bite by regulating the VEGF/ERK signal transduction pathway. METHODS Sixty SPF-grade healthy SD rats were selected and randomly divided into the normal control group, the Agkistrodon halys bite model group, the anti-Agkistrodon halys venom serum group, and the high -, medium -, and low-dose group of 717 Jiedu Decoction. Histopathological changes in the gastrocnemius muscle were observed using HE and Masson staining. Immunohistochemistry was used to detect the expression of CINC-1 and ICAM-1. Levels of VEGF, NGF, ERK, and IFN-γ in wound tissues were analyzed by ELISA, Western blot, and RT-qPCR, and ROS levels were measured with a biochemical assay kit. RESULTS the Agkistrodon halys bite model group showed marked tissue damage, while the serum and each dose groups of 717 JieDu Decoction exhibited varying degrees of improvement, with the high-dose group of 717 Jiedu Decoction closely resembling the normal control group. the Agkistrodon halys bite model group also showed increased collagen deposition and ROS levels, which were significantly reduced in the high-dose 717 JieDu Decoction group. CINC-1 and ICAM-1 expression was downregulated in the 717 JieDu Decoction groups, especially in the high-dose group of 717 Jiedu Decoction. VEGF, NGF, ERK, and IFN-γ were upregulated in the Agkistrodon halys bite model group, and both serum and high-dose 717 JieDu Decoction groups showed strong inhibitory effects on these factors. CONCLUSION 717 JieDu Decoction effectively reduces tissue fibrosis, oxidative stress, and inflammatory responses, the mechanism may be related to its inhibition of the VEGF/ERK signaling pathway, and the research provides a new target for the prevention and treatment of local tissue damage induced by Agkistrodon halysr venom. KEY WORDS: 717 JieDu Decoction; Agkistrodon halys bite; VEGF/ERK; local tissue injury