目的 基于網(wǎng)絡(luò)藥理學(xué)和實(shí)驗(yàn)驗(yàn)證探討瓜蔞皮注射液治療動(dòng)脈粥樣硬化的作用機(jī)制。方法 檢索瓜蔞皮注射液的化學(xué)成分報(bào)道，通過(guò)Swiss ADME和Swiss Target Prediction預(yù)測(cè)活性成分和靶點(diǎn)。通過(guò)OMIM、TTD、GeneCards數(shù)據(jù)庫(kù)獲得動(dòng)脈粥樣硬化的靶點(diǎn)，使用Venny 2.1軟件獲得瓜蔞皮注射液治療動(dòng)脈粥樣硬化的共有靶點(diǎn)。采用Cytoscape 3.6.1軟件構(gòu)建“藥物–成分–靶點(diǎn)”網(wǎng)絡(luò)，String、DAVID數(shù)據(jù)庫(kù)進(jìn)行蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)構(gòu)建，基因本體（GO）、京都基因和基因組百科全書(shū)（KEGG）富集分析。通過(guò)高脂飲食建立動(dòng)脈粥樣硬化小鼠模型，將小鼠隨機(jī)分為對(duì)照組、模型組、瓜蔞皮注射液（0.6、1.2 mL/kg）組，每組6只。瓜蔞皮注射液組分別im 0.6、1.2 mL/kg瓜蔞皮注射液，連續(xù)給藥3周。全自動(dòng)生化分析儀檢測(cè)小鼠血脂相關(guān)指標(biāo)，蘇木素–伊紅（HE）染色觀察主動(dòng)脈病理變化，蛋白質(zhì)免疫印跡（Western blotting）法檢測(cè)核心蛋白的表達(dá)。結(jié)果 篩選出芹菜素-7-O-β-D-葡萄糖苷、木犀草素、香草酸等21個(gè)瓜蔞皮注射液活性成分，共有靶點(diǎn)89個(gè)，核心靶點(diǎn)包括蛋白激酶B1（Akt1）、半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）、非受體酪氨酸激酶（SRC）等。GO功能分析獲得702個(gè)條目，KEGG富集獲得42條信號(hào)通路。動(dòng)物實(shí)驗(yàn)顯示，瓜蔞皮注射液可顯著降低血清總膽固醇（TC）、三酰甘油（TG）、低密度脂蛋白膽固醇（LDL-C）水平（P＜0.05、0.01）；HE染色顯示，瓜蔞皮注射液可減少斑塊在主動(dòng)脈管壁上附著；Western blotting顯示，瓜蔞皮注射液可顯著升高Caspase-3蛋白表達(dá)，降低p-SRC、p-Akt1蛋白表達(dá)（P＜0.05、0.01）。結(jié)論 瓜蔞皮注射液通過(guò)多個(gè)成分調(diào)控Caspase-3、p-SRC、p-Akt1等核心靶點(diǎn)，降低斑塊在主動(dòng)脈附著，進(jìn)而發(fā)揮治療動(dòng)脈粥樣硬化。

Objective To investigate the mechanism of action of Gualoupi Injection in treatment of atherosclerosis based on network pharmacology and animal experiments. Methods To search the chemical composition reports of Gualoupi Injection, and predict the active constituents and targets by Swiss ADME and Swiss Target Prediction. The targets of atherosclerosis were obtained by OMIM, TTD, and GeneCards databases, and the common targets of Gualoupi Injection for the treatment of atherosclerosis were obtained by Venny 2.1 software. Cytoscape 3.6.1 software was used to construct the “drug - component - target” network, String and DAVID databases were used to construct the PPI network, GO and KEGG enrichment analysis. The atherosclerosis mouse model was established by high-fat diet, and the mice were randomly divided into control group, model group, Gualoupi Injection (0.6, 1.2 mL/kg) group, with 6 mice in each group. Gualoupi Injection group was given 0.6 and 1.2 mL/kg of Gualoupi Injection, respectively, for 3 weeks. The relevant indexes of blood lipids were detected by automatic biochemical analyzer, the pathological changes of aorta were observed by HE staining, and the expression of core proteins was detected by Western blotting. Results 21 Active components of Gualoupi Injection, including apigenin-7-O-β-D-glucoside, luteolin, and vanillic acid were selected, with 89 targets. The core targets included Akt1, Caspase 3, and SRC. GO functional analysis obtained 702 items, KEGG enrichment obtained 42 signaling pathways. Animal experiments showed that the levels of TC, TG, and LDL-C in serum could be significantly reduced by Gualoupi Injection (P＜0.05, 0.01). HE staining showed that Gualoupi Injection could reduce plaque adhesion on the active vascular wall. Western blotting showed that Gualoupi Injection significantly increased the protein expression of Caspase 3, and decreased the protein expressions of p-SRC and p-Akt1 (P＜0.05, 0.01). Conclusion Gualoupi Injection regulates Caspase 3, p-SRC, p-Akt1 and other targets through multiple components, thereby reducing blood lipids and plaque attachment, and thus treating atherosclerosis.

R285.5；R972

鄭州大學(xué)第一附屬醫(yī)院橫向課題（K2020-0233）