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Objective To determine the content of total alkaloids and their components in roots, stems, and leaves of Berberis wilsonae, and investigate the effects of their extracts and components on the improvement of learning and memory ability and anti-oxidant activity of mice. Methods In this study, the total alkaloid content of roots, stems, leaves, and roots, stem extracts and components of B. wilsonae was determined by acid dye colorimetry. The HPLC method and conditions were established for determination jateorhizine, palmatine, and berberine. The memory impairment mice model was established with scopolamine, and the root and stem extracts and components of B. wilsonae were ig administrated for 30 d. Jumping platform test and water maze test were used to detect the effect of roots and stem extracts and components on learning and memory, and the levels of MDA and T-SOD in serum was determinated by kit method. Results The results showed that the latency of the mice in step-down test was significantly prolonged (P < 0.01), and the reduction of the number of errors was also significant (P < 0.01). To a certain extent, the place navigation time in water maze was shortened (P < 0.01), and the number of errors was reduced significantly (P < 0.05). Extracts and components can significantly increase the level of T-SOD (P < 0.05, 0.01) in mice, and reduce the level of MDA significantly (P < 0.05, 0.01). Conclusion The extracts and components of B. wilsonae had a certain effect on the improvement of learning and memory ability of mice, and the anti-oxidant capacity in vivo was also improved.
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Objective To establish a method for determining five isoquinoline alkaloids (including berberine,jatrorrhizine,jatrorrhizind,coptisine and palmatine) in Erhuang Xiaoyan tablets simultaneously.Methods Determination by reversed phase high performance liquid chromatography (RP-HPLC).Column:YMC-Pack ODS-AM (4.6 mm× 250.0 mm,5 μm);mobile phase:elution with 0.1％ (v/v) formic acid aqueous solution-acetonitrile mobile phase gradient;flow rate:0.4 mL/min;injection volume:10 μL;detection wavelength:345 nm;column temperature:27 ℃.Results Five of isoquinoline alkaloids in Erhuang Xiaoyan tablets were separated perfectly.The good linear relationships were obtained in the following ranges,1.526 4-5.342 4μg (r=0.999 9) for berberine,0.403 2-1.411 2 μg (r=0.999 6) for palmatine,0.309 7-1.083 9 μg (r=0.999 8) for coptisine,0.111 2-0.389 2 μg (r=0.999 8) for jatrorrhizine and 0.162 2-0.567 7 μg (r=0.999 6) for epiberberine.The recoveries were 98.69％,98.66％,98.67％,98.67％ and 98.67％,respectively.Conclusion The method possesses high feasibility,strong specificity,high accuracy and good reproducibility,which can be used for the quality control of Erhuang Xiaoyan tablets.
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Aim To study the biliary excretion characteristics of berberine,palmatine and jateorhizine in rat hepatocytes.Methods Berberine,palmatine and jateorhizine were incubated with the sandwich-cultured rat hepatocytes (SCRH) in standard Ca2+ buffer or Ca2+ free buffer.The accumulation of the three compounds under different conditions were measured by UPLC-MS/MS.The biliary excretion index and biliary clearance were calculated,and the effect of P-gp or Mrp2 inhibitor on the transport of three compounds was also investigated.Results While the incubation time increased,the accumulation of the three compounds also increased.There were obvious differences in accumulation of berberine,palmatine and jateorhizine in incubations treated with standard buffer and calcium-free buffer.The P-gp inhibitors ciclosporin A and verapamil could inhibit the biliary excretion of berberine,palmatine and jateorhizine.However,the Mrp2 inhibitors MK571 and probenecid had no effect on biliary excretion of the three compounds.Conclusions The biliary excretion of berberine,palmatine and jateorhizine is mainly through an active process.They are all the P-gp substrates other than Mrp2 substrates.
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OBJECTIVE: To evaluate the uptake characterizations of berberine, palmatine and jateorhizine through hepatic uptake model with rat primary hepatocytes. METHODS: Rat hepatocytes were isolated by Seglen's two-step method and cultured in suspension. The activity of transporters was assessed using rosuvastatin which is known as the substrate of the transporters. The tested drugs were incubated with rat hepatocytes at 4 or 37℃ for various periods of time and the drugs were determined using UPLC-MS/MS method to calculate the uptake amounts. The effect of cultural time and temperature on the hepatic uptake of these compounds were assessed. The kinetic parameters such as Km, Vmax and CLactive/CLuptake were calculated from concentration dependent uptake experiments. The effects of positive inhibitors on the uptake of berberine, palmatine and jateorhizine were also surveyed by pre-incubation of the inhibitors with the rat hepatocytes, followed by co-incubation with berberine, palmatine and jateorhizine. RESULTS: The hepatic uptake of berberine, palmatine and jateorhizine increased along with the increase of concentration at 37℃ while the change of hepatic uptake was very little at 4℃. The value of CLactive/CLuptake was 85.26%, 74.90% and 57.74% for berberine, palmatine and jateorhizine. Ibuprofen, digoxin, glycyrrhizin and prazosin which were the known inhibitors of transporters could remarkably reduce the hepatic uptake of berberine and palmatine, respectively. Glycyrrhizin and prazosin could reduce the uptake of jateorhizine. CONCLUSION: The transport of berberine, palmatine and jateorhizine into hepatocytes is mainly through an active process. Berberine and palmatine are the substrates of Oatp1a1, Oatp1a4, Oatp1b2 and Oct1, and the uptake of jateorhizine might be associated with Oatp1b2 and Oct1.