基于网络药理学和动物实验探讨养血荣筋丸治疗糖尿病微血管病变的作用*

作者：靳 冉1，岳 枫1，裴海鸾1，张晓笛2，徐 意2，李晋生2

单位：1.北京同仁堂科技发展股份有限公司制药厂，北京 100079； 2.北京同仁堂股份有限公司科学研究所，北京 100079

引用：引用：靳冉，岳枫，裴海鸾，张晓笛，徐意，李晋生.基于网络药理学和动物实验探讨养血荣筋丸治疗糖尿病微血管病变的作用[J].中医药导报,2025,31(3):37-44,88.

DOI：10.13862/j.cn43-1446/r.2025.03.007

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摘要：

目的：基于网络药理学和动物实验探讨养血荣筋丸治疗糖尿病微血管病变（DMA）的作用。方法：利用TCMSP、ETCM和PubChem数据库筛选养血荣筋丸有效成分并预测靶点，通过GeneCards、OMIM、PharmGkb和DrugBank数据库获得DMA相关疾病靶标基因，利用Cytoscape软件建立“药物成分-疾病靶点网络”，基于共有靶点使用String数据库构建蛋白质互作网络（PPI），利用David6.7数据库，进行GO功能和KEGG通路富集分析。采用高脂饲料联合尾静脉注射链脲佐菌素（STZ）法建立糖尿病（DM）大鼠模型后进行药物干预。给药10周后检测血糖、足部皮肤血流再灌注量、血液流变学、血清晚期糖基化终产物（AGEs）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）水平，HE染色观察眼球、肾脏和足底皮肤组织形态，过典酸雪夫（PAS）染色观察视网膜毛细血管管腔形态，免疫组化法检测肾脏组织中转化生长因子-β1（TGF-β1）的表达情况。结果：筛选养血荣筋丸活性成分103个，PPI网络中关键交集靶点95个，核心基因包括IL-1β、TNF、白细胞介素-10（IL-10）、白细胞介素-6（IL-6）等核心靶点7个，涉及晚期糖基化终产物及其受体（AGE-RAGE）和核因子-κB（NF-κB）等信号通路。动物实验结果显示，在给药后不同时间点，模型组大鼠血糖水平均高于对照组（P<0.01），羟苯磺酸钙组、养血荣筋丸高剂量组、养血荣筋丸中剂量组及养血荣筋丸低剂量组大鼠血糖水平与模型组比较，差异均无统计学意义（P>0.05）。给药干预结束后，模型组大鼠血流灌注量（Flux）低于对照组（P<0.01），全血黏度高于对照组（P<0.01）；模型组大鼠眼球视网膜明显增厚，肾小球基底膜样物质增多，管状基底膜增厚，足底皮肤毛细血管增生明显；模型组大鼠血清AGEs、IL-1β、TNF-α水平和肾脏中TGF-β1表达水平均高于对照组（P<0.01）；羟苯磺酸钙组、养血荣筋丸高剂量组、养血荣筋丸中剂量组及养血荣筋丸低剂量组大鼠Flux高于模型组（P<0.01）；羟苯磺酸钙组大鼠低、中、高切全血黏度低于模型组（P<0.01）；养血荣筋丸中、低剂量组大鼠中、高切全血黏度低于模型组（P<0.01或P<0.05）；羟苯磺酸钙组、养血荣筋丸高剂量组、养血荣筋丸中剂量组及养血荣筋丸低剂量组大鼠视网膜增厚情况改善，足底上皮细胞变性、真皮层胶原纤维萎缩变性程度降低；羟苯磺酸钙组大鼠血清IL-1β及TNF-α水平低于模型组（P<0.01或P<0.05）；养血荣筋丸高剂量组大鼠血清AGEs、IL-1β及TNF-α水平和肾脏中TGF-β1水平低于模型组（P<0.05或P<0.01）；养血荣筋丸中、低剂量组大鼠血清IL-1β水平均低于模型组（P<0.01)。结论：养血荣筋丸治疗DMA具有多成分、多靶点的特性，可能通过调节AGE-RAGE和NF-κB信号通路，改善DMA大鼠肢体末端微循环，减轻受累脏器损伤，发挥治疗作用。

关键词：糖尿病微血管病变；养血荣筋丸；网络药理学；AGE-RAGE；NF-κB信号通路

Abstract：

Objective: To investigate the effect of Yangxue Rongjin pill on diabetic microangiopaopathy (DMA) based on network pharmacology and animal experiments. Methods: TCMSP, ETCM and PubChem databases were used to screen the effective components of Yangxue Rongjin pill and predict the target. The DMA-related disease target genes were obtained from GeneCards, OMIM, PharmGkb and DrugBank databases. The "drug ingredient-disease target network" was established using Cytoscape software, and a String database was used to build a PPI network based on common targets. The enrichment analysis of GO function and KEGG pathway was carried out using David6.7 database. Diabetic rat model was established by high fat diet combined with streptozotocin (STZ) injection through tail vein, and then drug intervention was performed. After 10 weeks of administration, blood glucose level,vascular reperfusion volume of foot skin, blood rheology, serum  advanced glycation end products (AGEs), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels were detected. HE staining was used to observe the morphology of eye, kidney and plantar skin tissues, and periodic acid Schiff (PAS) staining was used to observe the morphology of retinal capillary lumens. Immunohistochemistry was used to detect the expression of transforming growth factor-β1 (TGF-β1) in kidney tissues. Results: Totally 103 active ingredients of Yangxue Rongjin pill were screened, and 95 key intersection targets were identified in PPI network. Core genes included IL-1β, TNF, interleukin-10 (IL-10), interleukin-6 (IL-6) and other core targets, which were involved in advanced glycation end products - receptor for advanced glycation end products (AGE-RAGE) and nuclear factor-kappa B (NF-κB) signaling pathways. The experimental results revealed that at various time points after administration, the model group showed higher blood glucose values than control group (P<0.01). Compared with the model group, there was no statistically significant difference in blood glucose levels between the calcium dobesilate group, high-dose Yangxue Rongjin pill group, medium-dose Yangxue Rongjin pill group, and low-dose Yangxue Rongjin pill group (P>0.05). After the administration intervention, the model group showed lower blood flow perfusion (Flux) than control group (P<0.01), while higher whole blood viscosity than control group (P<0.01). The retina of the eyes of rats was significantly thickened in model group, and the basement membrane-like substances in the glomeruli increased in model group. The tubular basement membrane thickened, and the capillary proliferation in the plantar skin was obvious in in model group. The model group showed higher levels of AGEs, IL-1β and TNF-α in serum and expression of TGF-β1 in the kidneys than control group (P<0.01). The calcium dobesilate group, high-dose Yangxue Rongjin pill group, medium-dose Yangxue Rongjin pill group, and low-dose Yangxue Rongjin pill group showed higher Flux values than model group (P<0.01). The calcium dobesilate group showed lower low, middle and high shear rates than model group (P<0.01). The medium-dose Yangxue Rongjin pill group, and low-dose Yangxue Rongjin pill group showed lower middle and high shear rates than model group (P<0.01 or P<0.05). The thickening of the retina in rats improved in calcium dobesilate group, high-dose Yangxue Rongjin pill group, medium-dose Yangxue Rongjin pill group, and low-dose Yangxue Rongjin pill group, and the degeneration of plantar epithelial cells and the atrophy and degeneration of collagen fibers in the dermis were reduced. The calcium dobesilate group showed lower contents of IL-1β and TNF-α than  model group (P<0.01 or P<0.05). The high-dose Yangxue Rongjin pill group showed lower contents of AGEs, IL-1β and TNF-α and TGF-β1 in the kidneys, than model group (P<0.05 or P<0.01). The medium-dose Yangxue Rongjin pill group, and low-dose Yangxue Rongjin pill group showed lower contents of IL-1β than model group (P<0.01). Conclusion: Yangxue Rongjin pill shows the characteristics of multiple components and multiple targets in the treatment of DMA, which may improve the microcirculation of the distal ends of limbs in DMA rats by regulating the AGE-RAGE and NF-κB signaling pathways, alleviating the damage to affected organs, and thus exerting therapeutic effects.

Key words：diabetic microangiopathy; Yangxue Rongjin pill; network pharmacology; AGE-RAGE; NF-κB signaling pathway

发布时间：2025-12-14

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