Integrated serum pharmacochemistry, pharmacokinetics, and network analysis to explore active components of BuShao Tiaozhi Capsule on hyperlipidemia

Abstract

BuShao Tiaozhi Capsule (BSTZC), a novel drug in China, has been used to treat hyperlipidemia (HLP) in clinical practice for many years. Despite our previous studies suggesting that BSTZC can treat HLP, there is a lack of a rapid and systematic method to explore its active components. Therefore, in this study, we aimed to investigate the active components and mechanisms of BSTZC in treating HLP by integrating serum pharmacology, pharmacokinetics, network analysis, and experimental validation. We first established UPLC fingerprints, calibrated 23 common peaks, and identified 13 common peaks, and the similarity was greater than 0.99 for 10 batches. A total of nine metabolites from BSTZC were identified in serum and considered as PK markers. The pharmacokinetic parameters of the PK markers were compared between the control group and the model group through the pharmacokinetics study to determine the dynamic changes of representative components in rats. Compared with the control group, the C_max and AUC_0→t of OXY, IVT, IVL, and KPF-3-G were significantly higher (P< 0.05); the AUC_0→∞ of OXY, PN, and IVT was significantly higher (P< 0.05); and the t_1/2 of IVT, SA, and KPF-3-G was significantly different (P< 0.05). In vivo experiments showed that BSTZC and its active components could effectively alleviate lipid metabolism disorders and liver injury, with obvious lipid-lowering effects. Further studies showed that BSTZC alleviated HLP by inhibiting the PI3K/Akt signaling pathway, which was consistent with the results of the network analysis study. Our results revealed the active components and mechanisms of BSTZC in the treatment of HLP, which could provide useful information to guide the clinical application of BSTZC.

Keywords: BuShao Tiaozhi Capsule; hyperlipidemia; network analysis; pharmacokinetics; serum pharmacochemistry.

FIGURE 1

BSTZ quality control. (A) UPLC fingerprints of BSTZ samples from 10 batches and controls. (B) A total of 13 peaks were identified by standards. Peaks 1, 4, 6, 7, 10, 12, 13, 15, 16, 17, 18, 19, and 21 were gallic acid, oxypaeoniflorin, chlorogenic acid, paeoniflorin, ferulic acid, salicylic acid, vitexin, isovitexin, neoandrographolide, isoviolanthin, astragalin, narcissoside, and neoandrographolide.

FIGURE 2

Total ion chromatograms of the BSTZC in the positive/negative ion mode (A, B), blank serum in the positive/negative ion mode (C, D), and rat serum after oral administration of BSTZC in the positive/negative ion mode (E, F).

FIGURE 3

Chromatograms of nine tested components ((A) mixed standard solution, (B) blank plasma, (C) serum sample after oral administration of BSTZC, and (D) blank solution. 1: OXY; 2: PN; 3: IVT; 4: IVL; 5: SA; 6: KPF-3-G; 7: NCS; 8: APG-7-G; 9: NAG).

FIGURE 4

Serum biochemical parameters (A) and HE staining of liver sections (B).

FIGURE 5

Mean plasma concentration–time curves of nine components in control and HLP model groups (1: OXY; 2: PN; 3: IVT; 4: IVL; 5: SA; 6: KPF-3-G; 7: NCS; 8: APG-7-G; 9: NAG).

FIGURE 6

Venny diagram (A), PPI network diagram (B), and bubble diagram (C).

FIGURE 7

Lipid-lowering effect of BSTZC and its active components on triton WR-1339-induced HLP mice. (A) Biochemical analyses of serum TC, TG, LDL-c, ALT, and AST. (B) mRNA levels of PI3KCA, AKT, PTGS2, and TNF in mice liver. The results are represented as mean ± SD of six mice in each group. ^## p < 0.01 vs. the control group, **p < 0.01 < 0.01 vs. the model group.