四妙勇安汤降低动脉粥样硬化炎症因子高敏C反应蛋白的拆方研究

Prescription-splitting study on Simiao Yong’an Decoction in reducing atherosclerotic inflammatory factor high sensitivity C-reactive protein

关键词:  四妙勇安汤  金银花  动脉粥样硬化  炎症  高敏-C反应蛋白

Key words:  Simiao Yong’an Decoction  honeysuckle  atherosclerosis  inflammation  high sensitivity C-reactive protein

摘要:

[目的] 明确四妙勇安汤复方中降低动脉粥样硬化炎症因子高敏C反应蛋白（hs-CRP）的核心药物，为临床精准用药提供实验依据。[方法] 采用单味药拆方实验，通过高脂饲料喂养联合注射脂多糖和维生素D3制备炎症模型，30只SD大鼠随机分为6组，包括空白对照组、模型组、金银花组、当归组、玄参组、甘草组，每组5只，动态监测药物干预后大鼠血清 hs-CRP 水平的变化；通过构建RAW264.7巨噬细胞与 HepG2肝细胞条件培养基共培养炎症模型和直接共培养炎症模型，双重验证核心药物对C反应蛋白（CRP）的调控作用；通过右侧颈总动脉套管手术联合高脂饲料喂养制备动脉粥样硬化模型，20只ApoE^-/-小鼠随机分为4组，包括模型组、他汀组、秋水仙碱组、金银花组，每组5只，并以C57BL/6J小鼠普通饲料喂养，作为正常对照组，探索核心药物潜在机制。[结果] 拆方动物实验显示，与空白对照组相比，模型组hs-CRP水平显著增高（P<0.05或P<0.01），与模型组相比，金银花组第4天、第7天及第10天hs-CRP水平持续下降，呈现时间依赖效应，并至第10天显著低于模型组（P<0.05）；其余当归组、玄参组、甘草组差异无统计学意义（P>0.05）；转移与共培养细胞模型中，金银花标醇提取物组与水提取物组均能显著降低CRP水平（P<0.01），其效果与阳性对照秋水仙碱组相近（P>0.05）。机制研究显示，与模型组相比，金银花组中核因子-κB（NF-κB，P<0.001）、白细胞介素-1β（IL-1β，P<0.001）、白细胞介素-6（IL-6，P<0.001）的表达水平显著降低，其效果与阳性对照秋水仙碱组相近（P>0.05）。[结论] 金银花是四妙勇安汤降低动脉粥样硬化炎症因子hs-CRP的关键药物，其作用与抑制NF-κB/IL-1β/IL-6/CRP通路相关，为动脉粥样硬化抗炎治疗的中医药临床应用和药物研发提供方向。

Abstract:

[Objective] To identify the core component of Simiao Yong’an Decoction responsible for reducing high sensitivity C-reactive protein（hs-CRP），a key inflammatory factor in atherosclerosis，and provide experimental evidence for precise clinical medication. [Methods] Single-herb disassembling experiments were performed. An inflammation model was established by feeding a high-fat diet combined with injections of lipopolysaccharide（LPS）and vitamin D3. Thirty Sprague-Dawley（SD）rats were randomly divided into 6 groups（5 rats per group）：blank control group，model group，Lonicera japonica group，Angelica sinensis group，Scrophularia ningpoensis group，and Glycyrrhiza uralensis group. Serum hs-CRP levels in rats were dynamically monitored after drug intervention. To double-verify the core herb’s regulatory effect on CRP，two inflammation models were constructed：a conditioned medium co-culture model of RAW264.7 macrophages and HepG2 hepatocytes，and a direct co-culture inflammation model. An atherosclerosis model was established via right common carotid artery cannulation combined with high-fat diet feeding. Twenty ApoE^-/- mice were randomly assigned to 4 groups（5 mice per group）：model group，statin group，colchicine group，and Lonicera japonica group. C57BL/6J mice fed a regular diet served as the normal control group. The potential mechanism of the core herb was explored. [Results] In the herb-disassembling animal experiment：Compared with the blank control group，the model group exhibited a significant increase in hs-CRP levels（P<0.05 or P<0.01）. Relative to the model group，the Lonicera japonica group showed a continuous decrease in hs-CRP levels on days 4，7，and 10（presenting a time-dependent effect），with a significantly lower level than the model group on day 10（P<0.05）. No significant differences were observed in the Angelica sinensis，Scrophularia ningpoensis，or Glycyrrhiza uralensis groups（P>0.05）. In the transfer and co-culture cell models：Both the ethanol extract and aqueous extract of Lonicera japonica significantly reduced CRP levels（P<0.01），and their effects were comparable to those of the positive control（P>0.05）. Mechanistic studies：Compared with the model group，the Lonicera japonica group showed markedly decreased expression levels of NF-κB（P<0.001），IL-1β（P<0.001），and IL- 6（P<0.001）；its effects were similar to those of the colchicine group（P>0.05）. [Conclusion] Lonicera japonica is the key herb in Simiao Yong’an Decoction for reducing the inflammatory factor hs-CRP in atherosclerosis，and its action is associated with the inhibition of the NF-κB/IL-1β/IL-6/CRP pathway. This study provides directions for the clinical application of traditional Chinese medicine and drug development in anti-inflammatory therapy for atherosclerosis.