Journal of Ethnopharmacology (Volume 260, 5 October 2020, 113044)

Ethnopharmacological relevance: San-Ye-Tang-Zhi-Qing formula (SYTZQ) is an effffective prescription for the treatment of pre-diabetes disorders of glycolipid metabolism in type 2 diabetes mellitus (T2DM). It consists of  five Chinese herbs including Mori Folium, Nelumbinis Folium, Crataegi Folium, Salviae Miltiorrhizae Radix et Rhizoma and Paeoniae Radix Rubra.

Aim of the study: This study was aimed to reveal the pharmacological mechanism of pharmacokinetic target components of SYTZQ for the treatment of T2DM.

Materials and methods: A rapid, precise and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to quantify simultaneously nuciferin, vitexin-4″-O-glu-coside, vitexin-2″-O-rhamnoside, paeoniflflorin and rosmarinic acid in rat plasma after oral administration of SYTZQ. The network pharmacology was used to analyze the effffect of the compounds absorbed into the blood of  SYTZQ on T2DM. The effffects of paeoniflflorin, nuciferine and rosmarinic acid on adipogenic difffferentiation werevalidated in vitro experiments.

Results: The separation was performed on an ACQUITY UHPLC HSS T3 column (2.1 mm × 100 mm, 1.7 μm) using acetonitrile and 0.1% (v/v) formic acid in water as the mobile phase in gradient elution. The calibration curves of fifive analytes showed good linearity (r ≥ 0.9991) with the lower limits of quantifification (LLOQ)  between 0.3 and 5.0 ng/mL. The recoveries and matrix effffects of fifive analytes ranged from 81.1% to 113%. The RSDs of inter-day and intra-day precision were all within 13.7%. The validated method was successfully appliedto the pharmacokinetic study of fifive ingredients after oral administration of SYTZQ to rat. 39 major targets and22 candidate pathways of fifive compounds absorbed into the blood of rats after administration of SYTZQ were identifified and successfully constructed a compound-target-disease-pathway network. It was confifirmed that  paeniforin, nuciferine and rosmarinic acid could decrease the adipogenicity difffferentiation in vitro experiments. Conclusions: The pharmacokinetic parameters indicated that the fifive components (nuciferin, vitexin-4″-O-glu-coside, vitexin-2″-O-rhamnoside, paeoniflflorin and rosmarinic acid) were absorbed and eliminated quickly in vivo. These fifive absorbed components were associated with 22 pathways, including insulin resistance, regulation  of lipolysis in adipocytes, PI3k/AKT-, TNF-, cAMP- and cGMP-PKG-signaling pathway. Paeoniflflorin, nuciferine and rosmarinic acid have the effffect of inhibiting adipocyte difffferentiation. This study could provide more re-ference for quality control, and provide a fifirm basis for evaluating the clinical effiffifficiency of SYTZQ.