花椒抗炎镇痛网络药理学分析及实验验证研究

Network pharmacological analysis and experimental verification of anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium

基于网络药理学及炎症疼痛小鼠模型探讨花椒的抗炎镇痛药理作用及机制。通过TCMSP数据库筛选花椒的活性成分、PharmMapper数据库获得花椒活性成分的靶基因,通过检索DrugBank、TTD、DisGeNET药物疾病靶点数据库收集炎症疼痛相关靶基因,并通过Cytoscape 3.7.0软件构建“活性成分-基因-疾病”网络。结果发现,花椒有5个潜在活性成分,预测到29个相关联的靶基因,在DrugBank、TTD、DisGeNET数据库中共检索筛选到132个炎症疼痛相关靶基因,“活性成分-基因-疾病”网络分析结果表明磷脂酰肌醇3激酶催化亚基γ亚型蛋白(PIK3CG)可能是花椒抗炎镇痛作用的关键靶点。通过二甲苯或角叉菜胶诱导的炎症小鼠模型,醋酸或福尔马林诱导的疼痛小鼠模型探讨花椒挥发油部位提取物和二氯甲烷部位提取物的抗炎镇痛药效。结果显示,花椒挥发油部位、二氯甲烷部位提取物能降低二甲苯诱导的小鼠耳肿胀和角叉菜胶诱导的小鼠足肿胀,减少醋酸诱导的小鼠扭体反应次数和福尔马林诱导的小鼠在第二时相的舔足时间,表明花椒具有抗炎镇痛的作用。进一步通过Western blot法检测蛋白表达,对花椒抗炎镇痛机制进行初步探讨。结果表明,花椒提取物可以降低PIK3CG、磷酸化核因子κB(p-NF-κB)和磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)蛋白的表达水平。该研究体现了花椒抗炎镇痛多成分、多靶点作用的特点,为花椒的抗炎镇痛研究提供药效学依据,为进一步的作用机制研究提供参考。

To explore the mechanism of anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium based on network pharmacology and inflammatory or pain mouse models. The effective components of Zanthoxyli Pericarpium were screened out by TCMSP database. And their potential corresponding targets were predicted by PharmMapper software. The possible targets relating to inflammation and pain were mainly collected through DrugBank, TTD and DisGeNET databases. The "active ingredient-gene-disease" network diagram was constructed by Cytoscape 3.7.0 software. The network pharmacology results showed 5 potential effective compounds, which were related to 29 targets;132 targets relating to inflammation and pain were screened out in the DrugBank, TTD and DisGeNET databases. The network analysis results indicated that the phosphatidylinositol 3-kinase catalytic subunit gamma isoform(PIK3 CG) gene may be the key to the anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium. The anti-inflammatory and analgesic effects of essential oil extract and dichloromethane extract of Zanthoxyli Pericarpium were explored through the mouse model of inflammation induced by xylene or carrageenan and the mouse model of pain induced by acetic acid or formalin. The experimental results showed that essential oil extract and dichloromethane extract of Zanthoxyli Pericarpium could reduce xylene-induced ear swelling and carrageenan-induced paw swelling and decrease the number of writhing responses in mice induced by acetic acid and the licking foot time of mice in phase Ⅱ induced by formalin. Western blot results showed that Zanthoxyli Pericarpium extract could inhibit the expressions of PIK3 CG, phosphonated nuclear factor kappaB(p-NF-κB) and phosphonated p38(p-p38 MAPK) protein. The present study showed the anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium through multiple components and targets, so as to provide a pharmacodynamic basis for the study of Zanthoxyli Pericarpium and its mechanism.