Isolation and Identification of Chemical Constituents from Rhizomes of Dendropanax proteus / 中国实验方剂学杂志

The chemical constituents from 95% ethanol extract of Dendropanax proteus rhizomes and their anti-inflammatory activities were investigated. These compounds in 95% ethanol extract of D. proteus rhizomes were isolated and purified by silica gel column chromatography, medium-pressure liquid chromatography, preparative liquid chromatography, etc. Their structures were elucidated based on the spectral data and physicochemical properties. All the compounds were tested for their ability to inhibit lipopolysaccharide (LPS) induced nitric oxide production in the murine microglia BV2 cell line. Nine compounds were isolated from the ethyl acetate fraction of 95% ethanol extract of D. proteus rhizomes, and identified as (-)-syringaresinol (1), (+)-(7S,8S)-1',4-dihydroxy-3,3',5'-trimethoxy-7',8,9'-trinor-8,4'-oxyneoligna-7,9-diol (2), erythro-guaiacylglycerol-β-O-4'-coniferyl ether (3), threo-guaiacylglycerol-β-O-4'-coniferyl ether (4), coniferyl alcohol (5), 7-O-ethylguaiacylglycerol (6), vanillin (7), syringaldehyde (8), and excoecanol B (9). Compounds 2 and 4 showed neuritis inhibitory activity against microglial inflammation factor, their half inhibitory concentrations (IC50) were 5.85, 7.29 μmol ·L-1, respectively. Compounds 1-6,8-9 are isolated from this plant for the first time, compounds 2 and 4 exhibit the potent inhibitory activity.

Research progress of pharmacological effects of mulberroside / 中国药理学通报

Mulberroside, a glycosylated stilbene, is the main bioactive constituent of white mulberry root-bark ( Sangbaipi) . It is widely used in many famous traditional Chinese medicine pre-scriptions to treat gout, arthritis, and rheumatism through pur-ging diuresis and relieving edema. In recent years, the pharma-cological activity of mulberroside has drawn extensive attention. With the utilization of the techniques and methods of modern pharmacology, a variety of biological activity and pharmacologi-cal effects of mulberroside are discovered gradually. The recent progress in the research on pharmacological effects of mulberro-side was reviewed in this paper to provide reference for the fur-ther development and comprehensive utilization.

Regulation of P-glycoprotein gene expression by PKC/NF-κB-PXR signaling pathway / 药学学报

P-glycoprotein (P-gp), an ATP binding cassette protein, plays a major role in efflux transport of drugs and xenobiotics due to its abundant expression on several barriers. This study aimed to investigate the potential role of PKC/NF-κB-PXR signaling pathway in modulation of P-gp gene expression in human colon adenocarcinoma LS174T. The effect of PMA on MDR1 luciferase activity was investigated by PXR-MDR1 dual luciferase reporter gene assay. Real-time qPCR assay and Western blot analysis were used to study the gene expression of P-gp and NF-κB, respectively. Compared to the vehicle-treated group, PMA statistically decreased P-gp luciferase activity, mRNA expression and protein expression. Moreover, PMA treatment yielded a significant and dose-dependent increase in RelA/p65 translocation to nucleus. Meanwhile, a remarkable increase of the pho-IκBα status was observed in LS174T cells after treatment with PMA (1-100 nmol·L-1). In addition, knockdown of PKCα, NF-κB or PXR can significantly attenuate PMA-induced P-gp suppression.These results suggested that PKC/NF-κB-PXR signaling pathway might play crucial roles in modulation of P-gp gene expression.

OATP1B1 in drug-drug interactions between traditional Chinese medicine Danshensu and rosuvastatin / 药学学报

The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharma­cokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*1a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of Cmax, AUC0-t, AUC0-∞ were increased about 123%, 194% and 195%, by Danshensu in rats, while the CLz/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol·L-1 Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol·L-1. The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvas­tatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94) % and (63.61 ± 3.94) %, respectively, by Danshensu at 1 and 10 μmol·L-1. While transport of rosuvastatin was reduced by (8.22 ± 2.40) % and (11.56 ± 3.04) % and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATP1B1. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.

OATP1B1 in drug-drug interactions between traditional Chinese medicine Danshensu and rosuvastatin / 药学学报

The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharmacokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of C(max0, AUCO(0-t), AUC(0-∞) were increased about 123%, 194% and 195%, by Danshensu in rats, while the CL z/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol x L(-1) Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol x L(-1). The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvastatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94)% and (63.61 ± 3.94)%, respectively, by Danshensu at 1 and 10 μmol x L(-1). While transport of rosuvastatin was reduced by (8.22 ± 2.40)% and (11.56 ± 3.04)% and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATPJBI. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.

Effect of Ursolic Acid on Breast Cancer Resistance Protein-mediated Transport of Rosuvastatin In Vivo and Vitro / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate whether ursolic acid can inhibit breast cancer resistance protein (BCRP)-mediated transport of rosuvastatin in vivo and in vitro.</p><p><b>METHODS</b>Firstly, we explored the pharmacokinetics of 5-fluorouracil (5-FU, a substrate of BCRP) in rats in the presence or absence of ursolic acid. Secondly, we studied the pharmacokinetics of rosuvastatin in rats in the presence or absence of ursolic acid or Ko143 (inhibitor of BCRP). Finially, the concentration-dependent transport of rosuvastatin and the inhibitory effects of ursolic acid and Ko143 were examined in Madin-Darby Canine Kidney (MDCK) 2-BCRP421CC (wild type) cells and MDCK2-BCRP421AA (mutant type) cells.</p><p><b>RESULTS</b>As a result, significant changes in pharmacokinetics parameters of 5-FU were observed in rats following pretreatment with ursolic acid. Both ursolic acid and Ko143 could significantly affect the pharmacokinetics of rosuvastatin. The rosuvastatin transport in the BCRP overexpressing system was increased in a concentration-dependent manner. However, there was no statistical difference in BCRP-mediated transport of rosuvastatin betweent the wild type cells and mutant cells. The same as Ko143, ursolic acid inhibited BCRP-mediated transport of rosuvastatin in vitro.</p><p><b>CONCLUSION</b>Ursolic acid appears to be a potent modulator of BCRP that affects the pharmacokinetic of rosuvastatin in vivo and inhibits the transport of rosuvastatin in vitro.</p>

Effect of Danshen on pharmacokinetics of Rosuvastatin in rats / 第二军医大学学报

Objective To study the effect of Danshen on the pharmacokinetics of Rosuvastatin in rats. Methods Parallel control method was applied in this study. Twelve male rats weighing (230 ± 15) g were evenly randomized into two groups: rosuvastatin (80 mg/kg) group and Rosuvastatin plus Danshen (400 mg/kg) group. Blood samples (0. 3 ml each) were collected from the orbital vein at 0. 5, 1, 1. 5, 3, 5, 8, 10, 12 and 24 h after drug administration. LC-MS was used to determine the plasma concentrations of Rosuvastatin. The pharmacokinetic parameters of the two groups were compared and statistical analysis was performed using SPSS12. 0 software. Results When combined with Danshen, the Cmax, AUC0-t, and AUCo-∞ of Rosuvastatin were increased by 60. 81%, 88. 00% and 82. 92%,and T1/2 and CLz/F were reduced by 33. 21% and 21. 51%, respectively. Conclusion Danshen can affect the pharmacokinetic properties of Rosuvastatin in rats.

Effect of OATP1B1 521T --> C heterogenesis on pharmacokinetic characterstics of rosuvastatin in Chinese volunteers / 药学学报

This study is to report the effect of OATP1B1 gene mutation in the 521T --> C in Chinese human on the pharmacokinetics of rosuvastatin and guide the reasonable clinical application of rosuvastatin by the feature of genetic polymorphism of OATP1B1. Plasma samples were determined with LC-MS: the analyte and internal standard pitavastatin were both analyzed by MS in the ESI, m/z was 480.0 for rosuvastatin and 420.0 for the IS, separately. Genotyping of OATP1B1 was determined with the method of polymerase chain reaction--amplification refractory mutation system targeted at 40 healthy volunteers and showed that there were 7 subjects with 521T --> C mutant, accounting to 17.5% of total and wild type homozygote accounted to 82.5%. It was found that there were significant differences between OATP1B1 mutation in the 521T --> C and wild type homozygote for rosuvastatin pharmacokinetic process in Chinese human. In contrast to OATP1B1 wild type group, OATP1B1 mutation group's absorption degree increased, elimination process decreased. The OATP1B1 mutation should be noted for guiding the reasonable application of rosuvastatin during its clinical use.

Quantification of pentoxyverine citrate in human plasma by LC-ESI/MS method and its application / 药学学报

A rapid and sensitive liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI/ MS) method for quantification of pentoxyverine citrate in human plasma has been developed and applied for the bioequivalence and pharmacokinetics study. After extracted from plasma samples with ethyl acetate, analysis was performed in selected ion monitoring (SIM) mode with a positive electrospray ionization (ESI) interface with a mobile phase consisted of methanol and water (0.4% glacial acetic acid and 4 mmol x L(-1) ammonium acetate, 43 : 57, v/v). The linear concentration range of the calibration curves was 1.0-160.0 ng x mL(-1) for pentoxyverine citrate, inter- and intra-precision (RSD) was less than 12.5%, accuracy (RE) was in +/- 13.5% and absolute recovery was more than 80%. The method was proved simple, rapid, sensitive, specific and suitable for pharmacokinetic and bioequivalence study of Yufenweilin capsule containing pentoxyverine citrate.

Occurrence of supernumerary upper incisor teeth in Pax6-/- mouse fetuses / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the occurrence of supernumerary upper incisor teeth in Pax6-/- mouse fetuses and to provide a model to explore the role of Pax6 in the upper incisor development and the mechanism of supernumerary teeth involving Pax6.</p><p><b>METHODS</b>Twenty Pax6-/- mouse fetuses of strain DEBA were isolated on E18.5 (embryonic day). The fetuses were sectioned serially in coronal plane and stained with haematoxylin and erosion, then the presence of supernumerary teeth in the upper anterior area was examined histologically, and also the number, morphology and structure of lower incisor germs and the first and second molar germs in the maxilla and mandible were observed histologically. Eighteen E18.5 mouse fetuses of strain DEBA with Pax6+/+ genotype were used as control.</p><p><b>RESULTS</b>Of the 20 Pax6-/- fetuses examined, four possessed a single supernumerary tooth in the upper incisors' region. No supernumerary upper incisor teeth were observed in any of the 18 Pax6+/+ fetuses examined. In the regions of lower incisors and the first and second molars of the maxilla and mandible, no significant difference was observed between Pax6-/- and Pax6+/+ fetuses regarding the number, morphology and structure of tooth germs.</p><p><b>CONCLUSIONS</b>The results suggest that Pax6 played an important role in the development of upper incisor teeth in mice.</p>