Integrating Network Pharmacology and Component Analysis Study on Anti‐Atherosclerotic Mechanisms of Total Flavonoids of Engelhardia roxburghiana Leaves in Mice

Abstract

Engelhardia roxburghiana Wall. leaves are widely used to develop herbal teas in southeast of China due to medicinal use for diabetes mellitus and hyperlipidemia. Studies have demonstrated that the total flavonoids of E. roxburghiana leaves (TFER) exhibited regulatory effects on blood glucose and lipids. To clarify the active ingredients of TFER and their targets in treating atherosclerosis, the present study integrated chemical analysis, network pharmacology analysis and animal experimental studies. Firstly, high performance liquid chromatography‐mass spectrometry/mass spectrometry (HPLC/MS/MS) was utilized to identify components of TFER. Then, active ingredients were screened by oral bioavailability (OB) and drug‐likeness (DL) index. Thirdly, network was constructed to predict major targets of active ingredients against atherosclerosis. Finally, to verify parts of predicted signaling, Apoe−/− mice were used to develop atherosclerosis. Atherosclerotic plaques in aorta were evaluated by echocardiography. Then, serum lipids, target genes expressions in thoracic aorta were determined by qRT‐PCR and ELISA methods. Chemical analysis revealed 10 components in TFER sample, 7 of which acted as active ingredients, including naringenin, kaempferol, quercetin, isoengeletin, engeletin, astilbin and quercitrin. KEGG pathway analysis highly enriched in some inflammatory signalings, including NF‐κB signaling, Toll‐like receptor signaling and TNF signaling. The animal studies indicated that TFER reduced atherosclerotic plaques size in aorta and significantly decreased the serum lipids, down‐regulated NF‐κB signaling by decreasing mRNA level of NF‐κB p65 subunit, TNF‐α and VCAM‐1, as well as IL‐1β expressions in thoracic aorta, eventually alleviating atherosclerosis progression, which was in consistent with our prediction.

Total Flavonoids of Engelhardia roxburghiana Wall. Leaves Alleviated Foam Cells Formation through AKT/mTOR‐Mediated Autophagy in the Progression of Atherosclerosis

“…The content of total flavonoids was determined as mg rutin (R)/g ethanol extracts, which was 826.4 mg R/g extracts. 10 compounds, such as quercitrin, astilbin, engeletin, quercetin, kaempferol, were identified in TFER by accurate mass and tandem mass spectra, which were reported in our previous study [14] . The chemicals structure were provided in Section 3 of Supporting Information .…”

“…10 components, such as quercitrin, astilbin, engeletin, quercetin, kaempferol, were isolated from TFER sample and confirmed by accurate mass and tandem mass spectra. Additionally, network pharmacology analysis based on these components has predicted that some inflammation pathways (i. e., NF‐κB signaling, Toll‐like receptor signaling, TNF signaling, NOD‐like receptor signaling), cholesterol metabolism and insulin signaling such as FoxO signaling might be involved in the anti‐atherosclerotic effect of TFER [14] . Based on the prediction, we further confirmed the inhibition of TFER on NF‐κB signaling in vascular tissue of atherosclerotic mice, but detailed mechanism still remained unclear.…”

Hypoxia-Induced miR-137 Inhibition Increased Glioblastoma Multiforme Growth and Chemoresistance Through LRP6

“…U6 RT primer: 5′-AACGCTTCACGAATTTGCGT-3′; U6 PCR primers sense: 5′-CTCGCTTCGGCAGCACA-3′; antisense: 5′-TGGTGTCGTGGAGTCG-3′. The cDNAs were amplified by ChamQ SYBR ® qPCR Master Mix (Vazyme Biotech co., ltd, China) on a LightCycler480II Real-Time PCR thermocycler (Roche, Switzerland), and fold changes were calculated by relative quantification (2 -△△Ct ) ( 20 , 21 ).…”

Ten New Dammarane-Type Saponins with Hypolipidemia Activity from a Functional Herbal Tea—Gynostemma pentaphyllum

“…Recent reports demonstrated that some functional tea drinks may exert a positive effect on lowering blood lipid and cholesterol. For example, green tea such as Longjing tea in China has antioxidant and blood lipid lowering effects [4].…”

“…Recent reports demonstrated that some functional tea drinks may exert a positive effect on lowering blood lipid and cholesterol. For example, green tea such as Longjing tea in China has antioxidant and blood lipid lowering effects [ 4 ]. The constituents of these tea, including polyphenols, flavonoids, and volatile oil, etc., have several good bioactivities, such as the ability to lowering blood lipid levels and blood pressure, in addition to anti-oxidant protectives and inhibition of inflammation [ 5 , 6 ].…”