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基于TLR4/NF-κB/NLRP3通路探讨毛果鱼藤提取物抗痛风作用及机制
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作者单位:

1.广西民族大学 海洋与生物技术学院,广西多糖材料与改性重点实验室,南宁 530008;2.广西民族大学 化学化工学院,广西林产化学与工程重点实验室,南宁 530008;3.广西壮族自治区中医药研究院,南宁 530022

中图分类号:

R2-0;R22;R285.5;R289;R33

基金项目:

国家自然科学基金项目(81560713,81960164);广西高校中青年教师科研基础能力提升项目(2021KY0175)


Derris eriocarpa Extract Treats Gout via TLR4/NF-κB/NLRP3 Pathway
Author:
Affiliation:

1.Guangxi Key Laboratory for Polysaccharide Materials and Modifications,School of Marine Sciences and Biotechnology,Guangxi Minzu University,Nanning 530008,China;2.Guangxi Key Laboratory of Chemistry and Engineering of Forest Products,School of Chemistry and Chemical Engineering,Guangxi Minzu University,Nanning 530008,China;3.Guangxi Institute of Chinese Medicine and Pharmaceutical Science,Nanning 530022,China

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    摘要:

    目的 通过网络药理学分析结合体内、外实验验证,探究毛果鱼藤对痛风的改善作用及机制。方法 利用数据库获取毛果鱼藤的化学成分及其候选靶标,通过蛋白质互作和网络分析,筛选毛果鱼藤治疗痛风的关键网络靶标和潜在活性成分,并进行生物功能富集及通路分析;小鼠腹腔注射次黄嘌呤复制高尿酸血症模型,观察毛果鱼藤醇提取物对高尿酸血症及正常小鼠尿酸水平的影响;大鼠足跖注射微晶型尿酸钠结晶,复制痛风性炎症模型,观察毛果鱼藤醇提物对痛风性炎症的影响;二甲苯诱导急性炎症模型,观察毛果鱼藤醇提物的抗炎作用;热板法、扭体法观察毛果鱼藤的镇痛作用。脂多糖诱导RAW264.7细胞复制体外炎症模型,观察毛果鱼藤醇提物对相关信号通路的影响。结果 通过构建毛果鱼藤-成分-痛风疾病靶点网络及分子对接分析,获得毛果鱼藤治疗痛风的12个关键网络靶标和15个潜在活性成分,发现其可以通过大黄素、白桦脂酸、美迪紫檀素等成分作用于Toll样受体4(TLR4)、NOD样受体蛋白3(NLRP3)、核转录因子-κB(NF-κB)等靶点发挥抗高尿酸血症、抗炎、镇痛等作用。与正常组比较,模型组小鼠血清尿酸水平显著增高(P<0.01)、大鼠肿胀度显著增高(P<0.05,P<0.01)、小鼠耳廓肿胀度降低(P<0.05)、小鼠扭体次数显著减少(P<0.05,P<0.01)、小鼠热板痛阈显著提高(P<0.05),细胞中TLR4 mRNA水平、TLR4、NF-κB、NLRP3的蛋白表达及细胞上清中TLR-4、NF-κB水平显著升高(P<0.05,P<0.01)。与模型组比较,毛果鱼藤醇提物(20、10、5 g·kg-1)可以显著降低高尿酸血症小鼠血清尿酸水平(P<0.01),抑制大鼠足跖肿胀(P<0.05,P<0.01),降低细胞中TLR4 mRNA水平,降低细胞中TLR4、NF-κB、NLRP3的蛋白表达及降低细胞上清中TLR4、肿瘤坏死因子-α(TNF-α)、NF-κB、白细胞介素-6(IL-6)的水平(P<0.05,P<0.01)。结论 毛果鱼藤醇提物具有明显的抗痛风作用,作用机制可能与其对TLR4/NF-κB/NLRP3信号通路的影响有关。

    Abstract:

    Objective To explore the anti-gout effect and mechanism of Derris eriocarpa extract by network pharmacological analysis combined with in vivo and in vitro experimental verification.Method The chemical components and candidate targets of D. eriocarpa were obtained from the database. The key targets and potential active components of D. eriocarpa in the treatment of gout were screened by the protein-protein interaction analysis, and then the Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed for the key targets. A mouse model of hyperuricemia was established by intraperitoneal injection of hypoxanthine to observe the effect of D. eriocarpa alcohol extract on hyperuricemia. A rat model of gouty inflammation induced by the injection of microcrystalline sodium urate crystals into the foot and plantar was used to observe the effect of D. eriocarpa alcohol extract on gouty inflammation. A xylene-induced acute inflammation model was established to observe the anti-inflammatory effect of D. eriocarpa alcohol extract. The hot plate test and twisting test were performed to observe the pain-relieving effect of D. eriocarpa. The lipopolysaccharide (LPS)-induced RAW264.7 cells were used to study the anti-gout effect and mechanism of D. eriocarpa alcohol extract.Result A total of 12 key targets and 15 potential active components were obtained from the D. eriocarpa-component-gout target network. The emodin, betulinic acid, and medicarpin endowed D. eriocarpa with anti-hyperuricemia, anti-inflammatory, and pain-relieving effects by acting on Toll-like receptor 4 (TLR4), NOD-like reception protein 3 (NLRP3), and nuclear factor (NF)-κB. Compared with the control group, the model groups showed elevated serum uric acid level in mice (P<0.01), increased swelling degree of rats (P<0.05, P<0.01), alleviated the auricular swelling of mice (P<0.05), reduced the twisting times of mice (P<0.05, P<0.01), and increased the hot plate pain threshold (P<0.05). Moreover, the model group showed up-regulated mRNA level of TLR4 and protein levels of TLR4, NF-κB, and NLRP3 in cells (P<0.01), and elevated levels of TLR4 and NF-κB in the cell supernatant (P<0.05, P<0.01). Compared with the model group, the alcoholic extracts (20, 10, 5 g·kg-1) of D. eriocarpa lowered the serum uric acid level in hyperuricemic mice (P<0.01), inhibited foot and plantar swelling in rats (P<0.05, P<0.01), down-regulated the mRNA level of TLR4 and the protein levels of TLR4, NF-κB, NLRP3 in cells, and lowered the levels of TLR4, TNF-α, NF-κB, and IL-6 in cell supernatants (P<0.05, P<0.01).Conclusion D. eriocarpa alcohol extract may exert the anti-gout, anti-inflammatory, and pain-relieving effects by regulating the TLR4/NF-κB/NLRP3 signaling pathway.

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刘洪存,朱震鑫,卢月,肖阳,刘颖,韦桂宁,杨立芳.基于TLR4/NF-κB/NLRP3通路探讨毛果鱼藤提取物抗痛风作用及机制[J].中国实验方剂学杂志,2023,29(23):72~80

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  • 收稿日期:2022-12-09
  • 在线发布日期: 2023-11-06

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