Research on the Anti-hyperlipidemia Effect and Mechanism of Linderae Folium via UFLC-MS/MS Combined with Network Pharmacology and Experimental Validation

OBJECTIVE  To explore the potential mechanism of Linderae Folium on prevention and treatment of hyperlipidemia, UFLC-MS/MS was used to analyze the chemical components of Linderae Folium, then the main active components, targets and related pathways of Linderae Folium against hyperlipidemia were verified by the chemical composition, network pharmacology, molecular docking technology and animal experiments.

METHODS  The chemical constituents in the Linderae Folium were analyzed by UFLC-MS/MS, and the targets related to hyperlipidemia were predicted by network pharmacology, and a "compound-disease-target" network was constructed. Through GO function and KEGG analysis, the significantly enriched pathways were screened(P<0.05). The selected active ingredients and the key targets of hyperlipidemia were verified by molecular docking. Finally, animal experiments and RT-PCR were used to verify the efficacy and anti-hyperlipidemia key targets.

RESULTS There were 32 chemical components analyzed by qualitative analysis of Linderae Folium, including 8 alkaloids, 16 flavonoids and 8 lactones. A total of 13 active ingredients and 46 related gene targets were screened by network pharmacological study. To explore the potential mechanism of Lindera Folium on lipid-lowering effect based on GO and KEGG analysis, the results suggested that the mechanism was mainly involved in the following pathways: fluid shear stress and atherosclerosis, AGE-RAGE signaling pathway, TNF signaling pathway, adipocytokine signaling pathway and so on. Animal experiments showed that the 2 doses of alcohol extract of Linderae Folium could significantly regulate the plasma lipid, blood glucose, and liver lipid levels of hyperlipidemia mice. The results of RT-PCR showed that Lindera Folium could significantly upregulate the expression of scavenger receptor class B type Ⅰ (SR-BI) mRNA, and downregulate the level of acyl-coenzyme A: cholesterol acyltransferase(ACAT) mRNA in liver.

CONCLUSION Lindera Folium might have effective on alleviate hyperlipidemia, and it exerted multi-component, multi-target and multi-pathway synergistic effect. The top 5 potential active compounds were quercetin, kaempferol, canadine, boldine, and tetrahydrocolumbamine. These compounds might exert lipid-lowering mechanism by regulating atherosclerosis, adipocytokines and other signaling pathways, and participating in inflammatory response, fat metabolism, insulin resistance and vascular endothelial function.