School of Medicine, Qinghai University, Xining, Qinghai, 810001, China; Qinghai Provincial Key Laboratory of Tibetan Medicine Research and CAS Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, 810008, Xining, China.
Qinghai Provincial Key Laboratory of Tibetan Medicine Research and CAS Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, 810008, Xining, China.
J Ethnopharmacol. 2024 Dec 5;335:118657. doi: 10.1016/j.jep.2024.118657. Epub 2024 Aug 9.
Sea buckthorn (Hippophae rhamnoides L.) is a traditional Chinese medicinal and possesses a rich medical history in terms of treating gastric disorders, sputum and cough and liver injuries in oriental medicinal system. By reason of the complicated chemical constituents, the material basis and potential pharmacological mechanism of sea buckthorn acting on Non-alcoholic fatty liver disease (NAFLD) has not been clearly elucidated.
To explore the pharmacological efficacy and underlying mechanism of sea buckthorn triterpenoid acid enrichment (STE) in the treatment of NAFLD.
The approaches of Network pharmacology and experiment validation in vitro and in vivo were applied in this study. Firstly, targets of triterpenoid acid compounds and NAFLD were collected from databases. The crucial targets were screened by the construction of protein-protein interaction (PPI) network. Furthermore, the potential signaling pathways and targets affected by STE was predicted by GO together with KEGG enrichment analysis. Finally, the experiment validation was carried out through high-fat feeding NAFLD mice and lipid accumulation HepG2 cell model. Lipids and liver related biochemical indicators were determined, Oil Red O and H&E staining were employed to observe fat accumulation. In addition, the expression levels of proteins of key target and signal pathway anticipated in network pharmacology were detected to elaborated its action mechanism.
A total of 180 intersecting potential targets for enhancing NAFLD with STE were eventually identified. 6 key targets including AKT1, TNF, IL6, INS, JUN, STAT3 and TP53 were further identified and the AMPK-SREBP1 pathway was enriched. Animal experiment result showed that STE treatment could significantly reduce the levels of TG, TC, LDL-C, ALT and AST, increase the levels of HDL-C in serum, and improve lipid accumulation of epididymal fat and liver. The results of the lipid accumulation cell model indicated that STE and key compound oleanolic acid could diminish intracellular lipid levels of TG, TC, LDL-C and number of lipid droplets. Western blot results showed that the above beneficial effects could be achieved by regulating the expression of p-AMPK/AMPK, SREBP1, FAS, ACC, SCD protein.
This study confirmed the effect of STE on improving NAFLD and the potential action mechanism was involved in the regulation of the AMPK-SREBP1 pathway.
沙棘（Hippophae rhamnoides L.）是一种传统的中药，在东方医学体系中，具有治疗胃部疾病、痰和咳嗽以及肝损伤的丰富医学历史。由于其复杂的化学成分，沙棘对非酒精性脂肪性肝病（NAFLD）的作用的物质基础和潜在的药理机制尚不清楚。
探讨沙棘三萜酸富集物（STE）治疗非酒精性脂肪性肝病的药理作用及作用机制。
本研究采用网络药理学和体外及体内实验验证的方法。首先，从数据库中收集三萜酸化合物和非酒精性脂肪性肝病的靶点，通过构建蛋白质-蛋白质相互作用（PPI）网络筛选关键靶点，然后通过 GO 与 KEGG 富集分析预测 STE 影响的潜在信号通路和靶点。最后，通过高脂喂养非酒精性脂肪性肝病小鼠和脂肪堆积 HepG2 细胞模型进行实验验证。测定血脂和肝脏相关生化指标，用油红 O 和 H&E 染色观察脂肪堆积。此外，还检测了网络药理学中预测的关键靶点和信号通路的蛋白表达水平，以阐述其作用机制。
最终确定了 180 个与 STE 增强非酒精性脂肪性肝病相关的潜在靶点。进一步鉴定出 6 个关键靶点，包括 AKT1、TNF、IL6、INS、JUN、STAT3 和 TP53，并富集了 AMPK-SREBP1 通路。动物实验结果表明，STE 治疗可显著降低血清 TG、TC、LDL-C、ALT 和 AST 水平，升高 HDL-C 水平，改善附睾脂肪和肝脏的脂肪堆积。脂质堆积细胞模型的结果表明，STE 和关键化合物齐墩果酸可减少细胞内 TG、TC、LDL-C 和脂滴数量。Western blot 结果表明，上述有益作用可通过调节 p-AMPK/AMPK、SREBP1、FAS、ACC、SCD 蛋白的表达来实现。
本研究证实了 STE 改善非酒精性脂肪性肝病的作用，其潜在作用机制涉及调节 AMPK-SREBP1 通路。
