川芎-丹参配伍治疗脊髓损伤的网络药理学研究与验证

Chuanxiong Rhizoma-salviae Miltiorrhizae Radix et Rhizoma in the Treatment of Spinal Cord Injury:Based on Network Pharmacology and Animal Experiments

目的:基于网络药理学,预测川芎-丹参配伍治疗脊髓损伤的作用机制。方法:利用网络药理学工具,获得川芎-丹参治疗脊髓损伤的有效成分、核心靶点及信号通路。采用改良Allen重物打击法构建脊髓损伤模型,采用酶联免疫吸附试验(ELISA)测定各组大鼠活性氧(ROS)、丙二醛(MDA)水平;采用蛋白质印迹法检测各组大鼠脊髓组织中磷脂酰肌醇3激酶(PI3K)、p-PI3K、蛋白激酶B(AKT)、p-AKT蛋白表达。结果:化合物-靶点网络包含154个化合物和484个相应靶点;基因本体(GO)功能富集分析得447个条目;京都基因和基因组百科全书(KEGG)通路富集分析得到126条信号通路。动物实验结果表明,与假手术组比较,模型组ROS、MDA水平显著增高(P<0.01);p-PI3K、p-AKT蛋白表达显著下降(P<0.01);与模型组比较,中药高剂量组ROS、MDA水平显著下降(P<0.01);p-PI3K、p-AKT蛋白表达显著增高(P<0.01)。结论:川芎-丹参联合治疗脊髓损伤具有多成分、多靶点、多通路的优势,其作用机制可能与调控PI3K/AKT信号通路有关。

Objective:To predict the potential mechanism of Chuanxiong Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma in the treatment of spinal cord injury(SCI)based on network pharmacology.Methods:The effective components,core targets,and signaling pathways of Chuanxiong Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma in treating SCI were obtained by network pharmacology.The rat model of SCI was established by the modified Allen method.The levels of reactive oxygen species(ROS)and malondialdehyde(MDA)in the rats were determined by enzyme-linked immunosorbent assay(ELISA).The protein levels of phosphatidylinositol 3-kinase(PI3K),p-PI3K,protein kinase B(AKT),and p-AKT were determined by Western blotting.Results:The compound-target network consisted of 154 compounds and 484 corresponding targets.The gene ontology(GO)annotation predicted 447 GO terms.The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that the key targets were involved in 126 signaling pathways.The animal experiments showed that compared with the sham group,the modeling elevated the levels of ROS and MDA(P<0.01)and down-regulated the protein levels of p-PI3K and p-AKT.Compared with the model group,high-dose Chinese medicine lowered the levels of ROS and MDA(P<0.01)and up-regulated the protein levels of p-PI3K and p-AKT(P<0.01).Conclusion:Chuanxiong Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma can treat SCI in a multi-component,multi-target,and multi-pathway manner by regulating the PI3K/AKT signaling pathway.