Yongmei Guan

Sex dependent pharmacokinetics, tissue distribution and excretion of peimine and peiminine in rats assessed by liquid chromatography–tandem mass spectrometry

ETHNOPHARMACOLOGICAL RELEVANCEnFritillaria thunbergii Miq. has been traditionally used in China as antitussive and expectorant herbs, and newly used in the clinical treatment of leukemia in recent years.nnnAIMnTo investigate whether gender exerted a significant influence on the pharmacokinetics, tissue distribution and excretion of peimine and peiminine in Sprague-Dawley rats who were given a single oral administration of 4.25 g/kg Fritillaria thunbergii Miq. extract.nnnMATERIALS AND METHODSnSprague-Dawley rats were assigned into two groups based on the gender and orally administered 4.25 g/kg Fritillaria thunbergii Miq. extract for each individual pharmacokinetics, tissue distribution and excretion study.nnnRESULTSnCompared with female rats, peimine and peiminine were eliminated slowly from male rat plasma, and significant gender-related differences were observed in the pharmacokinetic parameters. Drug blood and tissue levels in male rats were significantly higher than the female counterparts except for several tissues, such as fat, muscle and skin. Gender also exerted a significant influence on the urine excretion but such effect was not observed in the feces excretion study.nnnCONCLUSIONSnGender exerted a significant influence on the pharmacokinetics, tissue distribution and urine excretion of peimine and peiminine. It is assumed that the sex-associated differences of peimine and peiminine in rats might be mainly result from sex-dependent expression and activity of drug metabolism enzymes and P-glycoprotein.

The protective and healing effects of Si Shen Wan in trinitrobenzene sulphonic acid-induced colitis

ETHNOPHARMACOLOGICAL RELEVANCEnSi Shen Wan is a traditional Chinese herbal medicine formula for the treatment of diseases with diarrhea, such as ulcerative colitis, allergic colitis and chronic colitis. To investigate the protective and healing effects of Si Shen Wan in the experimental colitis induced by trinitrobenzene sulphonic acid, and to furture explore its mechanism of action.nnnMATERIALS AND METHODSnRats with colitis treated with Si Shen Wan for 10 days. Colon wet weight, colon organ coefficient, colonic damage score and pathological change after trinitrobenzene sulphonic acid challenge were determined. The levels of MPO, MDA, GSH-PX, SOD and the expression of IL-4 and IL-10 mRNA in the colon were also measured.nnnRESULTSnAfter treatment, colon wet weight, colon organ coefficient and colonic damage score were lower than that in the control group (p<0.05). MDA and MPO concentrations in the inflamed colonic tissues were decreased remarkably in the treated groups compared with that in the control group (p<0.05). But SOD level, IL-4 and IL-10 mRNA expression in the inflamed colonic tissues were obviously increased.nnnCONCLUSIONSnIt is a potential path that protective effect of Si Shen Wan on impaired colonic mucosa rats with experimental colitis was accomplished by down-regulating the level of MDA and MPO, and up-regulating the level of SOD and the IL-4 and IL-10 mRNA expression in the colon mucosa.

Mechanistic studies of the transport of peimine in the Caco-2 cell model

Fritillaria thunbergii Miq. has been widely used in traditional Chinese medicine for its expectorant, antitussive, antiinflammatory and analgesic properties. Moreover, modern pharmacological studies have demonstrated that F. thunbergii Miq. has efficacy in the treatment of leukemia and cancers of the liver and cervix. Although the alkaloid, peimine, is largely responsible for these pharmacological effects, it has very low oral bioavailability. The aim of this study was to investigate the intestinal absorption of peimine in Caco-2 cell monolayers. Having demonstrated that peimine is non-toxic to Caco-2 cells at concentrations <200 μmol/L, the effect of peimine concentration, pH, temperature, efflux transport protein inhibitors and EDTA-Na2 on peimine transport were studied. The results show that peimine transport is concentration-dependent; that at pH 6.0 and 7.4, the Papp(AP-BL) of peimine is not significantly different but the Papp(BL-AP)) is; that both Papp(AP-BL) and Papp(BL-AP) at 4 °C are significantly higher than their corresponding values at 37 °C; that the P-glycoprotein (P-gp) inhibitors, verapamil and cyclosporin A, increase absorption of peimine; and that EDTA-Na2 has no discernible effect. In summary, the results demonstrate that the intestinal absorption of peimine across Caco-2 cell monolayers involves active transport and that peimine is a substrate of P-gp.

Scorpio and Scolopendra attenuate inflammation and articular damage in rats with collagen-induced arthritis

ETHNOPHARMACOLOGICAL RELEVANCEnScorpio and Scolopendra (SS) are two traditional Chinese medicines, which are generally used to treat rheumatoid arthritis (RA) in China. However, the mechanism is so far unclear. The purpose of this study was to explore the effects and mechanisms of SS in attenuating inflammation and joint injury in collagen-induced arthritis in rats.nnnMATERIALS AND METHODSnRA was induced in Wistar rats by injection of collagen, meanwhile, the rats were administrated daily either SS (0.4 g/kg, 0.2 g/kg, and 0.1 g/kg) or vehicle (physiological saline) for 42 days. The therapeutic effect of SS on RA was evaluated by pathological methods. T lymphocyte subsets and anti-collagen type II (CII) antibody were tested in peripheral blood. Tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4) and interleukin-10 (IL-10) were assessed in tissue homogenate of fresh joints.nnnRESULTSnThe inflammation and articular damage in SS powder-treated rats were attenuated significantly. In addition, SS powder was revealed to modulate the equilibrium of T lymphocyte subsets, down-regulate TNF-α and IL-1β, up-regulate IL-4 and IL-10, and significantly suppress the level of anti-CII antibody.nnnCONCLUSIONSnScorpio and Scolopendra, when used as a combination, reveal desirable effect for treatment of rheumatoid arthritis, and this beneficial effect may be accomplished through normalization of T lymphocyte subsets and the balance of Th1/Th2 cytokines.

Pharmacokinetics, tissue distribution and excretion of peimisine in rats assessed by liquid chromatography-tandem mass spectrometry.

Peimisine, the common ingredient of “zhebeimu” groups and “chuanbeimu” groups, is responsible for the expectorant and cough relieving effects. The aim of this study was to investigate the pharmacokinetics, tissue distribution and excretion of peimisine in male and female SD (Sprague-Dawley) rats by a rapid and sensitive LC-MS/MS (liquid chromatography-tandem mass spectrometry) method used carbamazepine as the internal standard after oral administration, carbamazepine was stated as an IS. The results showed that peimisine was slowly distributed, and eliminated from rat plasma and manifested linear dynamics in a dose range of 0.26–6.5xa0mg/kg. Tested by ANOVA, there were gender differences in the pharmacokinetic parameters of AUC0−t, AUC0−∞ among a single dose of 0.26, 1.3, 6.5xa0mg/kg (Pxa0<xa00.05). Drug blood and tissue levels in male rats were significantly higher than the female counterparts after oral administration, while both the males and the females showed high drug levels in spleen, kidney, lung, liver and heart. On the other hand, the peimisine levels that can be reached in uterus, ovary, testis and brain is low. The excretion study showed that little administered peimisine (<0.7xa0%) was recovered in the male and female bile. Approximately 13.46 and 15.05xa0% were recovered in female urine and feces, while 43.07 and 7.49xa0% were recovered in male urine and feces, respectively, which indicated that the major elimination route of male rats was urine excretion. In addition, there was significant differences in total cumulative excretive ratio of peimisine in feces (Pxa0<xa00.05) and no significant differences in the urine (Pxa0>xa00.05) at a dose of 1.3xa0mg/kg.

Analysis of the High-Performance Liquid Chromatography Fingerprints and Quantitative Analysis of Multicomponents by Single Marker of Products of Fermented Cordyceps sinensis

Fermented Cordyceps sinensis, the succedaneum of Cordyceps sinensis which is extracted and separated from Cordyceps sinensis by artificial fermentation, is commonly used in eastern Asia in clinical treatments due to its health benefit. In this paper, a new strategy for differentiating and comprehensively evaluating the quality of products of fermented Cordyceps sinensis has been established, based on high-performance liquid chromatography (HPLC) fingerprint analysis combined with similar analysis (SA), hierarchical cluster analysis (HCA), and the quantitative analysis of multicomponents by single marker (QAMS). Ten common peaks were collected and analysed using SA, HCA, and QAMS. These methods indicated that 30 fermented Cordyceps sinensis samples could be categorized into two groups by HCA. Five peaks were identified as uracil, uridine, adenine, guanosine, and adenosine, and according to the results from the diode array detector, which can be used to confirm peak purity, the purities of these compounds were greater than 990. Adenosine was chosen as the internal reference substance. The relative correction factors (RCF) between adenosine and the other four nucleosides were calculated and investigated using the QAMS method. Meanwhile, the accuracy of the QAMS method was confirmed by comparing the results of that method with those of an external standard method with cosines of the angles between the groups. No significant difference between the two methods was observed. In conclusion, the method established herein was efficient, successful in identifying the products of fermented Cordyceps sinensis, and scientifically valid to be applicable in the systematic quality control of fermented Cordyceps sinensis products.

Mechanisms of P-Glycoprotein Modulation by Semen Strychni Combined with Radix Paeoniae Alba

Semen Strychni has been extensively used as a Chinese herb, but its therapeutic window is narrowed by the strong toxicity of the compound, which limits its effectiveness. Radix Paeoniae Alba has been reported to reduce the toxic effects and increase the therapeutic effects of Semen Strychni, but the underlying mechanism remains unknown. This research aimed to explore the mechanism through which P-glycoprotein (P-gp) is modulated by Semen Strychni combined with Radix Paeoniae Alba in vitro. An MTT assay was used to study cytotoxicity in an MDCK-MDR1 cell model. Rh123 efflux and accumulation were measured to assess P-gp function. The expression levels of MDR1 mRNA and P-gp protein in MDCK-MDR1 cells were investigated. A P-gp ATPase activity assay kit was applied to detect the effect on P-gp ATPase activity. Semen Strychni combined with Radix Paeoniae Alba could induce P-gp-mediated drug transport by inhibiting brucine and strychnine transport in MDCK-MDR1 cells, enhancing the P-gp efflux function, upregulating the P-gp expression and MDR1 mRNA levels, and stimulating P-gp ATPase activity.

Investigation of HP- -CD Inclusion Compound Preparation Process for Pulsatilla Chinensis (Bunge) Regel Total Saponins

This paper used parallel test of single factor to investigate feed ratio of inclusion compound, drying method, the volatilization temperature of ethanol for Pulsatilla Chinensis (Bunge) Regel total saponins-HP-β-CD inclusion compound. The results showed that the best preparation process of inclusion compound was as following: the feed ratio of Pulsatilla total saponin extract and HP-β-CD was 1:1; After the Pulsatilla total saponins extract with ethanol solution was dropwise added to the HP-β-CD aqueous solution, the mixed solution was continuously stirred for 6h at 60C and was freeze-dried for 12h. The preparation process for Pulsatilla Chinensis (Bunge) Regel total saponins-HP-β-CD inclusion compound was simple and it is helpful to improve the drug dissolution.