FEBS LETTERS
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2018
·Vol. 593
·期 2
·175-186
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IF 2.999
Shengnan Yang,Yuan Zhang,Fukui Shen,Xiaoyao Ma,Man Zhang,Yuanyuan Hou,Gang Bai
Baicalin is one of the main flavonoids of the dried root of Scutellaria baicalensis Georgi and is reported to exert beneficial effects on the regulation of glucose/lipid metabolism. However, understanding its specific target and unique mechanism for improving glucose utilization is a challenge. In this paper, target fishing with a baicalin probe reveals that baicalin interacts with AKT. An immunofluorescence assay further demonstrates the colocalization of baicalin with AKT in the cytoplasm. A competitive test and virtual docking show that baicalin might bind to the pleckstrin homology domain of AKT. This specific binding hampers AKT membrane translocation, activates the phosphorylation of AKT on Ser473, induces the downstream glycogen synthase kinase 3β activation, and affects glycogen synthesis.
研究领域:
药理学
·
分子生物学
·
代谢
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产品关联
DOI
PubMed
高影响因子
FEBS Journal
·
2019
·Vol. 287
·期 9
·1816-1829
·
IF 4.739
Man Zhang,Xiaoyao Ma,Honglei Xu,Wenbo Wu,Xin He,Xiaoying Wang,Min Jiang,Yuanyuan Hou,Gang Bai
Swertiamarin (SW), a representative component in Flos Lonicerae Japonicae , has been reported to exert significant activity in preventing infections. In this research, we aim to clarify the details of SW and its target to explore SW's underlying anti-inflammatory mechanisms. An azide labeled SW probe was synthesized for protein target fishing, and the results demonstrated that AKT could be captured specifically. Immunofluorescence colocalization with AKT was implemented by a click reaction of the SW probe and alkynyl CY5. The result showed that AKT was one of the targets of SW. Then, a competitive combination experiment using a set of AKT inhibitors and a membrane translocation experiment confirmed that SW might target the pleckstrin homology (PH) domain of AKT. This specific binding directly deactivated the phosphorylation of AKT on both Ser473 and Thr308, which induced the dephosphorylation of IKK and NF-κB. Finally, proinflammatory cytokines (TNF-α, IL-6, and IL-8) were suppressed both in cells and in acute lung injury animal model by targeting AKT-PH domain. This study demonstrated that SW functions as a natural AKT inhibitor and presents significant anti-inflammatory activity by directly regulating the AKT-PH domain and inhibiting downstream inflammatory molecules.
研究领域:
药理学
·
免疫学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Journal of Functional Foods
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2019
·
IF 3.197
Yunbing Shen,Shengnan Yang,Xueyan Hu,Man Zhang,Xiaoyao Ma,Zhihua Wang,Yuanyuan Hou,Gang Bai
Puerarin, one of the major isoflavonoid compounds isolated from Gegen, is widely used to treat glucose and lipid disorders. Identifying the mechanism of puerarin’s action against lipid and glucose metabolism dysfunction is very important for its application and development. Based on the structure of puerarin, different chemical probes were synthesized to capture and identify puerarin’s protein target. A membrane translocation assay was conducted to identify the interaction between puerarin and target proteins. The results show that puerarin targets the PH domain of Akt1, inhibits Akt1′s transmembrane effect and activates Akt1. Activated Akt1 improves the phosphorylation or activity of downstream proteins, such as GSK-3β and PDE3, and ultimately regulates glucose and lipid metabolism. Puerarin, as a dietary supplement, could target the PH domain of Akt1 to activate the insulin pathway and improve insulin resistance, thereby assisting in the regulation of glucose and lipid metabolism disorders.
研究领域:
药理学
·
代谢紊乱
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
Journal of Functional Foods
·
2021
·
IF 4.451
Ge Fang,Chuanjing Cheng,Manqian Zhang,Xiaoyao Ma,Shengnan Yang,Xiaotao Hou,Jiagang Deng,Yuanyuan Hou,Gang Bai
Kaempferol and quercetin, as dietary flavonoids, are beneficial to diabetes risk reduction. Clarifying the mechanism of dietary flavonoids in improving glucose metabolism may meet the functional and nutritional requirements. In this paper, PCA revealed two glucuronides, kaempferol-3-glucuronide (K-3-G) and quercetin-3-glucuronide (Q-3-G), which were the primary metabolites. Target prediction, enzymatic activity, and AKT plasma translocation assays indicated that K-3-G and Q-3-G could target the AKT PH domain. Fluorescence spectrometry was used to measure the dissociation constants of K-3-G (K D = 27.6 μM) and Q-3-G (K D = 10.8 μM). Molecular docking provided interaction insights for K-3-G and Q-3-G with the AKT PH domain. K-3-G and Q-3-G activated the AKT/GSK3β phosphorylation, thereby improving glucose consumption. Our study demonstrates that the glucuronide metabolites of flavonoids directly target the AKT PH domain and activate AKT/GSK3β signal pathway, thereby improving glucose metabolism.
研究领域:
药理学
·
生物化学
·
营养学
·
代谢学
✓ 已验证
产品关联
DOI
高影响因子
Chinese Herbal Medicines
·
2023
·
IF 3.8
Jie Gao,Manqian Zhang,Xingwang Zu,Xue Gu,Erwei Hao,Xiaotao Hou,Gang Bai
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.
研究领域:
药理学
·
代谢学
·
细胞生物学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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