中南药学

目的 探讨清营汤治疗川崎病的作用机制。方法 采用网络药理学筛选清营汤治疗川崎病(KD)的关键靶点。并通过分子对接验证核心靶点与药物成分的结合活性。动物实验采用C57BL/6JSD小鼠随机分成空白对照组(NC)、川崎病模型组(KD)、清营汤低剂量组(QYT-L)、清营汤高剂量组(QYT-H)、阿司匹林组(ASC)，每组各10只。采用干酪乳酸杆菌(LCWE)造模，造模成功后，QYT-L组、QYT-H组、ASC组分别灌胃给予50 mg·kg^-1 QYT、100 mg·kg^-1 QYT及50 mg·kg^-1 ASC，NC组及KD组灌胃等体积磷酸盐缓冲液(PBS)，1次·d^-1，持续干预7 d。记录小鼠体重变化；心脏超声检查小鼠左心室射血分数与短轴收缩率；酶联免疫吸附法测定小鼠血清中白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α含量；蛋白免疫印迹法检测小鼠心脏冠脉组织中PI3K、Akt、p-Akt、TNF-α蛋白表达情况，以论证清营汤对KD模型小鼠的作用。结果 筛选获得清营汤活性成分与KD的关键核心靶点(包括AKT1、IL6、TNF、STAT3、MMP9、TGFB1等)，与关键核心成分木犀草素、黄连素、汉黄芩素、山柰酚、柳杉酚、槲皮素的分子对接结果显示普遍具有良好结合活性。动物实验结果显示，清营汤能有效改善KD小鼠的心肌功能指标，抑制炎症因子释放，降低TNF-α、PI3K、p-AKT蛋白表达。结论 清营汤可能是通过木犀草素、山柰酚、槲皮素等活性成分作用于IL6、AKT1、TNF-α、IL-1β、TGF-β1等靶点，调控TNF、PI3K-AKT信号通路与Th17细胞等介导一系列炎症反应，抑制炎性因子释放等方式发挥对KD的治疗作用。

Objective To determine the mechanism of Qingying decoction (QYT) for Kawasaki disease (KD).Methods Network pharmacology was used to screen the core targets of QYT for KD.Then the binding activity of the core targets with the drug components was verified by molecular docking.In the experiments,50 C57BL/6JSD mice were randomly divided into a blank control group (NC group),a Kawasaki disease model group (KD group),a QYT low-dose group (QYT-L group),a QYT high-dose group (QYT-H group),and an aspirin group (ASC group),with 10 mice in each group.The mice were modeled with lactobacillus casei (LCWE).After successful modeling,50 mg·kg^-1 QYT,100 mg·kg^-1 QYT and 50 mg·kg^-1 ASC were given by gavage in the QYT-L,QYT-H and ASC groups,respectively,while an equal volume of phosphate buffer solution was given by gavage in the NC group and the KD group,continuous intervention for 7 d.Changes in body weight of the mice were recorded.Cardiac ultrasonography was used to examine the left ventricular ejection fraction and short-axis fractional shortening.Enzyme-linked immunosorbent assay was used to determine the serum levels of interleukin (IL)-6,IL-1β,and tumor necrosis factor (TNF)-α in mice.Western blot was used to detect the expression of PI3K,Akt,p-Akt,and TNF-α proteins in the coronary tissues of mouse hearts.Results The core targets of QYT’s active ingredients,including AKT1,IL6,TNF,STAT3,MMP9,and TGFB1 were screened.The molecular docking with the active ingredients luteolin,berberine,wogonin,kaempferol,sugiol,quercetin showed generally good binding activities.The animal experiments showed that QYT effectively improved the myocardial function indexes,inhibited the release of inflammatory factors,and reduced the expression of TNF-α,PI3K and p-AKT in KD mice.Conclusion QYT achieves its therapeutic effect on KD through luteolin,kaempferol,and quercetin,etc.via the targets of IL6,AKT1,TNF-α,IL-1β,and TGF-β1 to modulate the TNF and PI3K-AKT signaling pathway with Th17 cells,etc.,which mediates a series of inflammatory responses and inhibits the release of inflammatory factors.