摘 要：目的：利用网络药理学结合体内实验，探讨潞党参治疗小鼠大脑局部缺血后再灌注损伤的潜在作用机制。方法：对C57BL/6小鼠大脑构建中动脉闭塞再灌注模型（Middle Cerebral Artery Occlusion/ Reperfusion, MCAO/R），建模成功后，将其分为假手术组、MCAO/R模型组、潞党参低、中、高剂量组，进行连续7天潞党参药物治疗；通过Zea Longa 5级评分法评估MCAO/R小鼠的神经功能损伤情况；利用网络药理学和分子对接技术预测党参治疗缺血性脑卒中（IS）的潜在作用机制和靶点，在此基础上利用Western blot验证预测靶点。结果：Zea Longa 5级评分法显示，与假手术组相比，模型组小鼠神经功能损伤严重（P<0.001），与模型组相比，潞党参高剂量组小鼠的神经功能有所改善（P<0.05）。网络药理学机制研究表明，党参抗IS的作用可能与氧化应激、脂代谢以及神经炎症通路相关。分子对接结果显示，党参中的苍术内酯Ⅲ和补骨脂内酯可能通过IL-2、MMP12和CXCL12基因靶点，改善IS后的神经功能。Western blot验证结果显示，与模型组相比，潞党参高剂量组分别对MMP12和CXCL12有着不同程度的抑制作用，且均有统计学意义；党参高剂量组对TNF-α的表达有明显的（P<0.01）的下调作用。结论：潞党参可能通过抑制神经炎症反应，来缓解IS/R的神经功能损伤，其发挥抗炎作用可能是通过下调MMP12、CXCL12和TNF-α的表达水平实现的。

Abstract : Objective: To investigate the potential mechanism of action of Ludangshen in treating local post-ischemic reperfusion injury in mouse brain using network pharmacology combined with in vivo experiments. Methods: A middle cerebral artery occlusion/ reperfusion (MCAO/R) model was constructed for the brains of C57BL/6 mice, and after successful modeling, they were divided into sham-operated group, MCAO/R model group, and Ludangshen low, medium and high dose groups, and treated with Ludangshen drugs for 7 consecutive days. The neurological impairment of MCAO/R mice were assessed by Zea Longa 5-point scale; network pharmacology and molecular docking techniques were used to predict the potential mechanism of action and targets of Ludangshen in the treatment of ischemic stroke. The predicted targets were validated by Western blot. Results: The Zea Longa 5-point scale showed that mice in the model group had severe neurological impairment compared with the sham-operated group (P<0.001), and mice in the high-dose group of Ludangshen had improved neurological function compared with the model group (P<0.05). Subsequent network pharmacological mechanism studies indicated that the anti-ischemic stroke effect of Codonopsis may be related to oxidative stress, lipid metabolism and neuroinflammatory pathways. Further molecular docking results showed that atractylenolide III and psoralen in Codonopsis may exert neurological improvement after ischemic stroke through IL-2, MMP12 and CXCL12 gene targets. The results of Western blot validation showed that compared with the model group, the high-dose group of Ludangshen had different degrees of inhibitory effects on MMP12 and CXCL12, respectively, and all of them were statistically significant; the high-dose group of Ludangshen had a significant (P<0.01) down-regulation effect on the expression of TNF-α. Conclusion: Ludangshen may alleviate neurological impairment after reperfusion of ischemic stroke by inhibiting neuroinflammatory response, and its exerting anti-inflammatory effect may be achieved by down-regulating the expression levels of MMP12, CXCL12 and TNF-α.

1. 山西省重点研发计划(国际科技合作)重大区域创新合作项目(201803D421006) 2.国家中医药管理局青年岐黄学者支持项目