Abstract
Berberrubine (BRB) is the primary metabolite of berberine (BBR) that has shown a stronger glucose-lowering effect than BBR in vivo. On the other hand, BRB is quickly and extensively metabolized into berberrubine-9-O-β-D-glucuronide (BRBG) in rats after oral administration. In this study we compared the pharmacokinetic properties of BRB and BRBG in rats, and explored the mechanisms underlying their glucose-lowering activities. C57BL/6 mice with HFD-induced hyperglycemia were administered BRB (50 mg·kg−1·d−1, ig) for 6 weeks, which caused greater reduction in the plasma glucose levels than those caused by BBR (120 mg·kg−1·d−1) or BRB (25 mg·kg−1·d−1). In addition, BRB dose-dependently decreased the activity of α-glucosidase in gut of the mice. After oral administration of BRB in rats, the exposures of BRBG in plasma at 3 different dosages (10, 40, 80 mg/kg) and in urine at different time intervals (0–4, 4–10, 10–24 h) were dramatically greater than those of BRB. In order to determine the effectiveness of BRBG in reducing glucose levels, we prepared BRBG from the urine pool of rats, and identified and confirmed it through LC-MS-IT-TOF and NMR spectra. In human normal liver cell line L-O2 in vitro, treatment with BRB or BRBG (5, 20, 50 μmol/L) increased glucose consumption, enhanced glycogenesis, stimulated the uptake of the glucose analog 2-NBDG, and modulated the mRNA levels of glucose-6-phosphatase and hexokinase. However, both BBR and BRB improved 2-NBDG uptake in insulin-resistant L-O2 cells, while BRBG has no effect. In conclusion, BRB exerts a stronger glucose-lowering effect than BBR in HFD-induced hyperglycemia mice. Although BRB significantly stimulated the insulin sensitivity and glycolysis in vitro, BRBG may have a greater contribution to the glucose-lowering effect because it has much greater system exposure than BRB after oral administration of BRB. The results suggest that BRBG is a potential agent for reducing glucose levels.
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Acknowledgements
This study was financially supported by the National Natural Science Foundation of China (No 81573495, 81530098), the Key Technology Projects of China “Creation of New Drugs” (No 2015ZX09501001), the Project for Jiangsu Province Key Lab of Drug Metabolism and Pharmacokinetics (No BM2012012) and the Project of the University Collaborative Innovation Center of Jiangsu Province (Modern Chinese Medicine Center and Biological Medicine Center).
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Supplementary information is available at website of Acta Pharmacologica Sinica.
Supplementary information
Figure S1
Ion peaks in rat plasma after p.o. BRB administration for 5 min. (DOC 171 kb)
Figure S2
MS and UV chromatograms of BRBG in urine samples. (DOC 224 kb)
Figure S3
Identification of different peaks in preparative system by LC-MS-PDA. (DOC 187 kb)
Figure S4
MS1 and MS2 fragments of target compound analyzed by LC-MS-IT-TOF system. (DOC 72 kb)
Table S1
Purity analysis of target compound. (DOC 32 kb)
Summary
This supplementary material contains five files: four figures and one table in total. Ion peaks of plasma, LC-MS-PDA analysis and MS fragments of target compound are shown in figures, respectively. Purity analysis of target compound is also listed in the table. (DOC 13 kb)
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Yang, N., Sun, Rb., Chen, Xl. et al. In vitro assessment of the glucose-lowering effects of berberrubine-9-O-β-D-glucuronide, an active metabolite of berberrubine. Acta Pharmacol Sin 38, 351–361 (2017). https://doi.org/10.1038/aps.2016.120
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DOI: https://doi.org/10.1038/aps.2016.120
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