目的 通過高效液相色譜-四極桿-靜電場軌道阱高分辨質(zhì)譜（HPLC-Q-Exactive Orbitrap MS）技術(shù)結(jié)合網(wǎng)絡(luò)藥理學(xué)和實驗驗證，探討金櫻子Rosa laevigata治療糖尿病腎病的藥效物質(zhì)及作用機制。方法 構(gòu)建糖尿病腎病大鼠模型考察金櫻子抗糖尿病腎病的藥效，并采用HPLC-Q-Exactive Orbitrap MS技術(shù)分析金櫻子在大鼠體內(nèi)的入血成分；基于已鑒定的入血成分，利用網(wǎng)絡(luò)藥理學(xué)預(yù)測金櫻子治療糖尿病腎病的潛在作用機制，并利用Cytoscape對網(wǎng)絡(luò)藥理學(xué)的結(jié)果進行進一步分析。采用分子對接實驗驗證核心活性成分與關(guān)鍵靶點的結(jié)合能力，同時借助糖尿病腎病大鼠模型驗證核心信號通路的調(diào)控作用。結(jié)果 金櫻子能顯著降低糖尿病腎病大鼠血糖（FBG）、24 h尿白蛋白（24 h-U-Alb）、尿素氮（BUN）、血肌酐（Scr）、總膽固醇（TC）、三酰甘油（TG）、低密度脂蛋白膽固醇（LDL-C）水平（P<0.05）及抑制白細(xì)胞介素-6(IL-6)、白細(xì)胞介素18(IL-18)、腫瘤壞死因子-α(TNF-α)等炎癥因子的分泌。鑒定出19個金櫻子入血成分，網(wǎng)絡(luò)藥理學(xué)分析顯示，金櫻子發(fā)揮作用的生物學(xué)過程主要涉及RNA聚合酶II正向調(diào)控轉(zhuǎn)錄、信號轉(zhuǎn)導(dǎo)、磷酸化、細(xì)胞增殖的正向調(diào)控、抑制凋亡過程、基因表達的正向調(diào)控等，通過STAT3、AKT1、PIK3R1、PIK3CA、GRB2、PTPN11、PIK3CB、EGFR等核心靶點，并調(diào)控癌癥、PI3K-Akt、癌癥中的蛋白聚糖、人巨細(xì)胞病毒感染、癌癥中的MicroRNAs、脂質(zhì)與動脈粥樣硬化等信號通路。分子對接驗證實驗表明，金櫻子關(guān)鍵活性成分與STAT3、AKT1、PIK3R1等核心靶點結(jié)合性較好。動物實驗進一步證實，金櫻子能顯著下調(diào)糖尿病腎病大鼠腎組織中PI3K、Akt與p-Akt蛋白的表達（P<0.05）。結(jié)論 金櫻子具有抗糖尿病腎病的藥效作用。其主要活性成分為沒食子酸、積雪草酸、羥基積雪草苷、δ-生育酚、芹菜素、甘草素、鞣花酸、12-羥基茉莉酸、原花青素B1。其抗糖尿病腎病分子機制可能與抑制炎癥因子分泌及調(diào)節(jié)PI3K-Akt信號轉(zhuǎn)導(dǎo)通路有關(guān)。

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

貴州省科技計劃項目(黔科合基礎(chǔ)-ZK[2023]一般416); 2023年度貴州中醫(yī)藥大學(xué)學(xué)術(shù)新苗項目(貴科合學(xué)術(shù)新苗[2023]-13號); 貴州省衛(wèi)生健康委科學(xué)技術(shù)基金項目(黔衛(wèi)健函[2024]24號,gzwkj2024-514); 貴州省基礎(chǔ)研究計劃(自然科學(xué))面上項目(黔科合基礎(chǔ)MS[2025]172); 貴州省中醫(yī)藥管理局中醫(yī)藥、民族醫(yī)藥科學(xué)技術(shù)研究課題(QZYY-2025-012); 貴州中醫(yī)藥大學(xué)藥用高分子材料研究中心(貴中醫(yī)ZX合字[2024]071號)