Mechanisms Underlying the Differences in the Pharmacokinetics of Six Active Constituents of Huangqi Liuyi Decoction between Normal and Diabetic Nephropathy Mouse Models

Abstract

The aim of this study was to explore the mechanisms underlying the differences in the pharmacokinetics of Huangqi Liuyi decoction extract (HQD) under physiological and pathological conditions. The roles of liver cytochrome P450 metabolic enzymes (Cyp450) and small intestinal transporters were also investigated. The cocktail probe drug method was used to investigate the effects of diabetic nephropathy (DN) and HQD on metabolic enzyme activity. The expression levels of liver Cyp450 metabolic enzymes (Cyp1A2, Cyp2C37, Cyp3A11, Cyp2E1, and Cyp2C11) and small intestinal transporters (breast cancer resistance protein (BCRP), P-glycoprotein (P-gp), organic cation transporters (OCTs), and multidrug resistance-associated protein (MRPs) were determined using western blot. Compared to normal mice, the expression of OCT1, OCT2, MRP1, and MRP2 was increased in DN mice, while that of P-gp and BCRP (P < 0.05 and P < 0.001) was inhibited. HQD inhibited expression of Cyp1A2 and Cyp3A11 and increased the expression of P-gp and BCRP in normal mice. In DN mice, HQD induced expression of BCRP and inhibited expression of Cyp2C37, Cyp3A11, OCT2, MRP1, and MRP2. The activity of each Cyp450 enzyme was consistent with changes in expression. The changes in pharmacokinetic parameters of HQD in DN might, in part, be secondary to decreased expression of P-gp and BCRP. HQD varied in regulating transporter activities between health and disease. These findings support careful application of HQD-based treatment in DN, especially in combination with other drugs.

Figure 1

The HPLC-MS/MS chromatogram of HQD. Blank sample (a), reference substance of six active ingredients in HQD (b), and HQD sample (c). (1) astragaloside IV, (2) calycosin-7-O-β-D-glucoside, (3) calycosin-glucuronide, (4) ononin, (5) formononetin, (6) glycyrrhizic acid, (7) puerarin (IS), (8) digoxin (IS).

Figure 2

The number of metabolites produced by specific probe substrates in liver microsomes (μmol/mL, x ± SD, n = 6). ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001 compared to the control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared to the model group.

Figure 3

Protein expression of CYP450 metabolic enzymes (Cyp1A2, Cyp2C37, Cyp3A11, Cyp2E1, and Cyp2C11) in different groups (n = 3). ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001 compared to the control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared to the model group.

Figure 4

Protein expression of small intestinal transporters (BCRP, P-gp, OCT1, OCT2, MRP1, and MRP2) in different groups (n = 3). ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001 compared to the control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared to the model group.