Glucuronic acid metabolites of phenolic acids target AKT-PH domain to improve glucose metabolism

IF 4.7 4区 医学 Q1 CHEMISTRY, MEDICINAL Chinese Herbal Medicines Pub Date : 2023-07-01 DOI:10.1016/j.chmed.2022.11.005
Jie Gao , Manqian Zhang , Xingwang Zu , Xue Gu , Erwei Hao , Xiaotao Hou , Gang Bai
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引用次数: 1

Abstract

Objective

Phenolic acids widely exist in the human diet and exert beneficial effects such as improving glucose metabolism. It is not clear whether phenolic acids or their metabolites play a major role in vivo. In this study, caffeic acid (CA) and ferulic acid (FA), the two most ingested phenolic acids, and their glucuronic acid metabolites, caffeic-4′-O-glucuronide (CA4G) and ferulic-4′-O-glucuronide (FA4G), were investigated.

Methods

Three insulin resistance models in vitro were established by using TNF-α, insulin and palmitic acid (PA) in HepG2 cells, respectively. We compared the effects of FA, FA4G, CA and CA4G on glucose metabolism in these models by measuring the glucose consumption levels. The potential targets and related pathways were predicted by network pharmacology. Fluorescence quenching measurement was used to analyze the binding between the compounds and the predicted target. To investigate the binding mode, molecular docking was performed. Then, we performed membrane recruitment assays of the AKT pleckstrin homology (PH) domain with the help of the PH-GFP plasmid. AKT enzymatic activity was determined to compare the effects between the metabolites with their parent compounds. Finally, the downstream signaling pathway of AKT was investigated by Western blot analysis.

Results

The results showed that CA4G and FA4G were more potent than their parent compounds in increasing glucose consumption. AKT was predicted to be the key target of CA4G and FA4G by network pharmacology analysis. The fluorescence quenching test confirmed the more potent binding to AKT of the two metabolites compared to their parent compounds. The molecular docking results indicated that the carbonyl group in the glucuronic acid structure of CA4G and FA4G might bind to the PH domain of AKT at the key Arg-25 site. CA4G and FA4G inhibited the translocation of the AKT PH domain to the membrane, while increasing the activity of AKT. Western blot analysis demonstrated that the metabolites could increase the phosphorylation of AKT and downstream glycogen synthase kinase 3β in the AKT signaling pathway to increase glucose consumption.

Conclusion

In conclusion, our results suggested that the metabolites of phenolic acids, which contain glucuronic acid, are the key active substances and that they activate AKT by targeting the PH domain, thus improving glucose metabolism.

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酚酸的葡萄糖醛酸代谢物靶向AKT-PH结构域,改善葡萄糖代谢
目的酚酸广泛存在于人类饮食中,具有改善葡萄糖代谢等有益作用。目前尚不清楚酚酸或其代谢产物是否在体内发挥主要作用。本研究对摄入量最大的两种酚酸咖啡酸(CA)和阿魏酸(FA)及其葡萄糖醛酸代谢产物咖啡-4′-O-葡萄糖醛酸(CA4G)和阿魏酸酯-4′-O葡萄糖醛酸(FA4G)进行了研究。方法应用TNF-α、胰岛素和棕榈酸分别建立HepG2细胞胰岛素抵抗模型。我们通过测量葡萄糖消耗水平,比较了FA、FA4G、CA和CA4G对这些模型中葡萄糖代谢的影响。通过网络药理学预测了潜在的靶点和相关途径。荧光猝灭测量用于分析化合物与预测靶标之间的结合。为了研究结合模式,进行了分子对接。然后,我们在PH-GFP质粒的帮助下进行了AKT-pleckstrin同源性(PH)结构域的膜募集测定。测定AKT酶活性以比较代谢物与其母体化合物之间的作用。最后,通过蛋白质印迹分析研究了AKT的下游信号通路。结果CA4G和FA4G在增加葡萄糖消耗方面比其母体化合物更有效。通过网络药理学分析预测AKT是CA4G和FA4G的关键靶点。荧光猝灭测试证实,与它们的母体化合物相比,这两种代谢物与AKT的结合更有效。分子对接结果表明,CA4G和FA4G的葡萄糖醛酸结构中的羰基可能在关键的Arg-25位点与AKT的PH结构域结合。CA4G和FA4G抑制了AKT PH结构域向膜的易位,同时提高了AKT的活性。蛋白质印迹分析表明,代谢产物可以增加AKT信号通路中AKT和下游糖原合成酶激酶3β的磷酸化,从而增加葡萄糖消耗。结论总之,我们的研究结果表明,含有葡萄糖醛酸的酚酸代谢产物是关键的活性物质,它们通过靶向PH结构域来激活AKT,从而改善葡萄糖代谢。
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文献相关原料
公司名称产品信息其他信息采购帮参考价格
索莱宝 Insulin from bovine pancreas
¥67.90~¥9000.00
希恩思 Palmitic acid sodium salt (PA)
¥19.00~¥7270.00
索莱宝 Metformin hydrochloride (Met)
¥2900.00~¥3138.90
上海源叶 Metformin hydrochloride (Met)
¥2900.00~¥3138.90
希恩思 Palmitic acid sodium salt (PA)
索莱宝 1,1'-Dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil)
上海源叶 Metformin hydrochloride
阿拉丁 Caffeic acid (CA)
>98.5%
阿拉丁 Ferulic acid (FA)
>98.5%
来源期刊
Chinese Herbal Medicines
Chinese Herbal Medicines CHEMISTRY, MEDICINAL-
CiteScore
4.40
自引率
5.30%
发文量
629
审稿时长
10 weeks
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