目的 通过高效液相色谱-四极杆-静电场轨道阱高分辨质谱（HPLC-Q-Exactive Orbitrap MS）技术结合网络药理学和实验验证，探讨金樱子Rosa laevigata治疗糖尿病肾病的药效物质及作用机制。方法 构建糖尿病肾病大鼠模型考察金樱子抗糖尿病肾病的药效，并采用HPLC-Q-Exactive Orbitrap MS技术分析金樱子在大鼠体内的入血成分；基于已鉴定的入血成分，利用网络药理学预测金樱子治疗糖尿病肾病的潜在作用机制，并利用Cytoscape对网络药理学的结果进行进一步分析。采用分子对接实验验证核心活性成分与关键靶点的结合能力，同时借助糖尿病肾病大鼠模型验证核心信号通路的调控作用。结果 金樱子能显著降低糖尿病肾病大鼠血糖（FBG）、24 h尿白蛋白（24 h-U-Alb）、尿素氮（BUN）、血肌酐（Scr）、总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）水平（P<0.05）及抑制白细胞介素-6(IL-6)、白细胞介素18(IL-18)、肿瘤坏死因子-α(TNF-α)等炎症因子的分泌。鉴定出19个金樱子入血成分，网络药理学分析显示，金樱子发挥作用的生物学过程主要涉及RNA聚合酶II正向调控转录、信号转导、磷酸化、细胞增殖的正向调控、抑制凋亡过程、基因表达的正向调控等，通过STAT3、AKT1、PIK3R1、PIK3CA、GRB2、PTPN11、PIK3CB、EGFR等核心靶点，并调控癌症、PI3K-Akt、癌症中的蛋白聚糖、人巨细胞病毒感染、癌症中的MicroRNAs、脂质与动脉粥样硬化等信号通路。分子对接验证实验表明，金樱子关键活性成分与STAT3、AKT1、PIK3R1等核心靶点结合性较好。动物实验进一步证实，金樱子能显著下调糖尿病肾病大鼠肾组织中PI3K、Akt与p-Akt蛋白的表达（P<0.05）。结论 金樱子具有抗糖尿病肾病的药效作用。其主要活性成分为没食子酸、积雪草酸、羟基积雪草苷、δ-生育酚、芹菜素、甘草素、鞣花酸、12-羟基茉莉酸、原花青素B1。其抗糖尿病肾病分子机制可能与抑制炎症因子分泌及调节PI3K-Akt信号转导通路有关。

Objective To investigate the active substances and molecular mechanisms of Rosa laevigata in the treatment of diabetic nephropathy through HPLC-Q-Exactive Orbitrap MS, network pharmacology, and experimental validation. Methods Firstly, a diabetic nephropathy rat model was established to evaluate the therapeutic effect of R. laevigata on diabetic nephropathy, and the blood components of R. laevigata were analyzed using HPLC-Q-Exactive Orbitrap MS technology. Secondly, based on the blood components of R. laevigata, network pharmacology was used to analyze the potential mechanisms of R. laevigata in treating diabetic nephropathy, and further analysis of the network pharmacology results was performed using Cytoscape. Lastly, molecular docking and a diabetic nephropathy rat model were employed to verify the key active components and core pathways. Results R. laevigata significantly reduced fasting blood glucose（FBG）, 24-h urinary albumin（24 h-U-Alb）, serum creatinine（Scr）, blood urea nitrogen（BUN）, triglyceride（TG）, total cholesterol（TC）, low-density lipoprotein cholesterol（LDL）（P＜0.05）, and inhibited the secretion of inflammatory cytokines such as interleukin-6（IL-6）, interleukin-18（IL-18）, and tumor necrosis factor-α（TNF-α）. Nineteen blood components of R. laevigata were identified. Network pharmacology analysis revealed that R. laevigata’s effects are primarily associated with various biological processes, including positive regulation of transcription by RNA polymerase II, signal transduction, phosphorylation, cell proliferation, inhibition of apoptosis, and gene expression. These effects are mediated through targets such as STAT3, AKT1, PIK3R1, PIK3CA, GRB2, PTPN11, PIK3CB, and EGFR, impacting signaling pathways related to cancer, PI3K-Akt, proteoglycans in cancer, human cytomegalovirus infection, microRNAs in cancer, and lipid and atherosclerosis. Molecular docking validation experiments further confirmed that the active components of R. laevigata exhibit strong binding affinities with key targets like STAT3, AKT1, and PIK3R1. Animal validation experiments showed that R. laevigata could down-regulate the expression of PI3K, Akt, and p-Akt proteins（P＜0.05）. Conclusion R. laevigata has a therapeutic effect on diabetic nephropathy. The main active components of R. laevigata are gallic acid, asiatic acid, asiaticoside, δ-tocopherol, apigenin, liquiritigenin, ellagic acid, 12-hydroxyjasmonic acid, and procyanidin B1. Its molecular mechanism may be related to the inhibition of inflammatory cytokine secretion and the regulation of the PI3K-Akt signaling pathway. The results of this study can provide experimental evidence for the development and application of functional foods derived from R. Laevigata.

R285.5

贵州省科技计划项目(黔科合基础-ZK[2023]一般416); 2023年度贵州中医药大学学术新苗项目(贵科合学术新苗[2023]-13号); 贵州省卫生健康委科学技术基金项目(黔卫健函[2024]24号,gzwkj2024-514); 贵州省基础研究计划(自然科学)面上项目(黔科合基础MS[2025]172); 贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2025-012); 贵州中医药大学药用高分子材料研究中心(贵中医ZX合字[2024]071号)