Modulation of Rat Hepatic CYP1A and 2C Activity by Honokiol and Magnolol: Differential Effects on Phenacetin and Diclofenac Pharmacokinetics In Vivo.

Honokiol (2-(4-hydroxy-3-prop-2-enyl-phenyl)-4-prop-2-enyl-phenol) and magnolol (4-Allyl-2-(5-allyl-2-hydroxy-phenyl)phenol) are the major active polyphenol constituents of Magnolia officinalis (Magnoliaceae) bark, which has been widely used in traditional Chinese medicine (Houpu Tang) for the treatment of various diseases, including anxiety, stress, gastrointestinal disorders, infection, and asthma. The aim of this study was to investigate the direct effects of honokiol and magnolol on hepatic CYP1A and 2C-mediated metabolism in vitro using rat liver microsomes and in vivo using the Sprague-Dawley rat model. Honokiol and magnolol inhibited in vitro CYP1A activity (probe substrate: phenacetin) more potently than CYP2C activity (probe substrate: diclofenac): The mean IC50 values of honokiol for the metabolism of phenacetin and diclofenac were 8.59 µM and 44.7 µM, while those of magnolol were 19.0 µM and 47.3 µM, respectively. Notably, the systemic exposure (AUC and Cmax) of phenacetin, but not of diclofenac, was markedly enhanced by the concurrent administration of intravenous honokiol or magnolol. The differential effects of the two phytochemicals on phenacetin and diclofenac in vivo pharmacokinetics could at least be partly attributed to their lower IC50 values for the inhibition of phenacetin metabolism than for diclofenac metabolism. In addition, the systemic exposure, CL, and Vss of honokiol and magnolol tended to be similar between the rat groups receiving phenacetin and diclofenac. These findings improve our understanding of CYP-mediated drug interactions with M. officinalis and its active constituents.

In vitro and in vivo evaluation of the effect of puerarin on hepatic cytochrome p450-mediated drug metabolism.

Puerarin (8-ß-D-glucopyranosyl-7-hydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) is a major pharmacological component of Puerariae Radix, the root of Pueraria lobata. We investigated the effect of puerarin on hepatic cytochrome P450-mediated drug metabolism in rats and humans. The in vitro cytochrome P450 inhibitory effect of puerarin in human and rat liver microsomes was evaluated using the following model cytochrome P450 substrates: phenacetin for CYP1A, diclofenac for CYP2C, dextromethorphan for CYP2D, and testosterone for CYP3A. The in vivo pharmacokinetics of intravenous and oral buspirone, a probe substrate for CYP3A, was studied with single simultaneous intravenous coadministration of puerarin in rats. In the in vitro cytochrome P450 inhibition study, the rate of disappearance of testosterone was significantly reduced in the presence of 10 µM PU, while that of other cytochrome P450 substrates was not significantly affected in both human and rat liver microsomes, suggesting that puerarin inhibits the in vitro hepatic CYP3A-mediated metabolism in the human and rat systems (IC50 = 15.5 ± 3.9 µM). After intravenous administration of buspirone with single simultaneous coadministration of intravenous puerarin at a dose of 10 mg/kg in rats, the total area under the plasma concentration-time curve from time zero to time infinity was increased while time-averaged total body clearance decreased. When buspirone was orally administered in rats with the 10 mg/kg intravenous puerarin coadministration, both total area under the plasma concentration-time curve from time zero to time infinity and the extent of absolute oral bioavailability were significantly increased. Therefore, results of the in vitro microsomal and in vivo pharmacokinetic studies suggest the possible inhibition of hepatic CYP3A-mediated drug metabolism by puerarin administration, potentially leading to metabolism-mediated herb-drug interactions with clinical significance.

Metabolic interactions of rosmarinic acid with human cytochrome P450 monooxygenases and uridine diphosphate glucuronosyltransferases.

In light of the widespread use of herbal medicines containing rosmarinic acid (RA) and limited literature available thereon, we investigated the metabolic interactions of RA with human cytochrome P450 monooxygenases (CYPs) and uridine diphosphate glucuronosyltransferases (UGTs). The involvement of selected enzymes (CYP1A2, CYP2C19, CYP2E1, CYP3 A4, UGT1A1, UGT1A6, and UGT2B7) in the metabolism of RA and the inhibitory effect of RA on the enzyme activity were comprehensively evaluated using human recombinant isozyme system. Additionally, concentration-dependent RA metabolism mediated by phase I enzymes (including CYPs) or UGT was investigated in human liver microsome (HLM) system. A significant disappearance of RA was observed in the seven CYP and UGT isoforms studied, indicating their possible involvement in the metabolism of RA. Based on Michaelis-Menten kinetics, the metabolism study using HLM suggests that the UGT system may have a higher capacity and lower affinity for the metabolism of RA than phase I enzyme (including CYP) systems. Moreover, RA weakly inhibited CYP2C9 and 2E1 activities with IC50 values of 39.6 and 61.0 µM, respectively, while moderately inhibiting UGT1A1, 1A6, and 2B7 with IC50 values of 9.24, 19.1, and 23.4 µM, respectively. By constructing Line weaver-Burk plots, the type of inhibition exhibited by RA on CYP and UGT activities was determined as follows: CYP2C19, mixed inhibition; CYP2E1, UGT1A1, UGT1A6, and UGT2B7, competitive inhibition. Based on the comparison of the IC50 and Ki values obtained in the current study with the previously reported plasma concentrations of RA after oral dosing in humans, it is suggested that RA may significantly inhibit the activities of the tested UGTs, rather than CYPs, in clinical settings. Thus, the present study could provide a basis for further studies on clinically significant interactions between UGT substrate drugs and herbal medicines containing RA.

Smooth muscle relaxation activity of an aqueous extract of dried immature fruit of Poncirus trifoliata (PF-W) on an isolated strip of rat ileum.

We demonstrated that an aqueous extract of dried immature fruit of Poncirus trifoliate (PF-W) produces relaxation of intestinal smooth muscle using the ileac strips of a rat. Furthermore, the underlying mechanism of its relaxant activity was investigated. PF-W was prepared using the standard extraction protocol. A 1.5 - 2 cm long rat ileac strip was placed in an organ bath with Tyrode's solution and smooth muscle contractility was recorded by connecting it to a force transducer. Various compounds were added to the organ baths, and changes in muscular contractility were measured. PF-W concentration-dependently induced relaxation of rat ileac strips that were contracted both spontaneously and via acetylcholine treatment. Various potassium channel blockers did not inhibit the relaxation by PF-W. No difference in the effect of PF-W was observed between ileac strips treated with low (20 mM) and high concentrations (60 mM) of KCl. PF-W inhibited the contraction of rat ileac strips induced by extracellular calcium. PF-W acts as a potent smooth muscle relaxant, implicating its possible action as a rapid acting reliever for abdominal pains and a cure for intestinal convulsion. Considering that PF-W also exhibits prokinetic activity, its use in various gastrointestinal disorders seems promising.

A survey for rhinitis in an automotive ring manufacturing plant.

We report findings regarding otolaryngologist-confirmed rhinitis, current exposure to MWF aerosols, fungi, and endotoxins for workers in a plant manufacturing automobile piston rings. Questionnaire data showed that 61.5% of 187 workers exhibited rhinitis-related symptoms. Rhinitis was confirmed in 99 of 115 workers whom were medically examined. Otolaryngologist-confirmed rhinitis was present in 10 of 19 grinding workers (52.6%), 67 of 142 production workers (47.2%), and 22 of 26 quality control (QC) workers (84.6%). These rates are much higher than the rates of rhinitis-related symptoms in automobile plants and other occupational settings and quite high even allowing for the common occurrence of rhinitis in the general population. We found that rhinitis could develop even in workers exposed to less than 0.5mg/m(3) MWF aerosol. The average exposure to fungi exceeded 10 x 10(3) CFU/m(3), a level higher than that reported for other automobile plants. Although we were unable to identify significant risk factors for rhinitis using only the physician-confirmed rhinitis cases, this study concludes that exposure to MWF aerosol, which would include microbes and metals, could contribute to a high occurrence of rhinitis in grinding and production workers. Forty-nine workers (63.6%) of 77 rhinitis patients in grinding and production operations were determined to handle synthetic MWF directly. For QC workers, for whom the prevalence of physician-confirmed rhinitis was highest, exposure to a low level of MWF aerosol, including specific microbe species we couldn't identify, bright light, dry air, and certain work characteristics during inspection are possible risk factors for development of rhinitis. Further studies including identification of fungi species should be conducted so a firm conclusion can be made regarding the development of rhinitis in QC manufacturing plant workers.

Association between use of synthetic metalworking fluid and risk of developing rhinitis-related symptoms in an automotive ring manufacturing plant.

The main objective of this study was to determine the association between synthetic metalworking fluid (MWF) and rhinitis-related symptoms. At a plant manufacturing piston rings for automobiles, we interviewed grinders (19) and manufacturing workers (142) in operations where synthetic or semisynthetic MWF is handled, and administrative office workers (44) regarding the principal symptoms of rhinitis (nasal stuffiness, runny nose, anosmia, nasal itchiness, rhinorrhea, headache, epistaxis, and post-nasal drip). In addition, we assessed the current exposure of workers handling MWF to MWF aerosols, fungi, and endotoxins. Logistic regression analysis was used to examine the association between MWF surrogates indicative of MWF exposure and each rhinitis-related nasal symptom. Odds ratios (ORs) and 95% confidence intervals were adjusted for sex, age, smoking habit, and duration of employment. Among grinders handling synthetic MWF, the frequency of complaints of the dominant symptoms was 66.7% for nasal stuffiness, 77.8% for anosmia, 77.8% for runny nose, and 50.0% for headache. These rates are quite high even allowing for the common occurrence of rhinitis in the general population. Twenty eight of 34 grinding and manufacturing workers (82.4%) sampled were exposed to MWF mist above the threshold limit of 0.2 mg/m(3) listed as a notice of intended change by the American Conference for Governmental Industrial Hygienists (ACGIH). The percentage of workers exposed to MWF mist >0.5 mg/m(3) was 17.6%. Most workers were exposed to fungi levels >103 CFU/m(3). All exposures to endotoxins were <50 EU/m(3). Logistic regression analysis found that use of synthetic MWF was significantly associated with excess risk of nasal stuffiness (OR 3.5), nasal itchiness (OR 2.0), and runny nose (OR 2.1). The use of semi-synthetic MWF had little or no impact on the risk of developing rhinitis-related nasal symptoms. Grinding workers handling synthetic MWF had an increased risk of nasal stuffiness (OR 7.9), anosmia (OR 23.2), nasal itchiness (OR 8.3), runny nose (OR 20.4), post nasal drip (OR 18.4), and headache (OR 7.4) compared to administrative workers. Synthetic MWF may play an important role in the development of the dominant symptoms of rhinitis. Further study is needed to establish the risk of rhinitis or rhinitis-related symptoms according to MWF type.

Avoidance therapy in reactive dye-induced occupational asthma: long-term follow-up.

BACKGROUND: Previous studies reported that early diagnosis and avoidance therapy are the most important factors for prevention of permanent lung impairment; however, few studies have evaluated the long-term prognosis of reactive dye-induced occupational asthma (RD-OA). OBJECTIVE: To evaluate the long-term outcomes of RD-OA. METHODS: Methacholine airway hyperresponsiveness (AHR) and lung functions were evaluated and compared in 26 patients with RD-OA at the time of diagnosis and after complete avoidance of the causative agents. Patients with continued (n = 13) or remitted (n = 6) AHR were further monitored for up to a mean +/- SD of 8.7 +/- 1.8 years. RESULTS: The AHR resolved in 10 (38%) of 26 patients a mean +/- SD of 2.2 +/- 1.3 years after complete avoidance of RDs; however, prebronchodilator forced expiratory volume in 1 second (FEV1) values were not different. Levels of IgE specific to the RD-human serum albumin complex were markedly decreased at first follow-up in 5 RD-atopic patients from whom paired serum samples were compared (P = .02). The AHR disappeared in an additional 5 patients and improved in 4 by the second follow-up. The FEV1 values also improved compared with diagnosis and first follow-up levels. Favorable prognosis was associated with early diagnosis of RD-OA and complete avoidance of the causative agent. No association was found with smoking history, latent periods, the presence of RD specific IgE, baseline provocation concentration that caused a decrease in FEV1 of 20%, or FEV1. CONCLUSIONS: Early diagnosis and avoidance therapy are the most important prognostic factors in RD-OA. The AHR and lung function of patients with RD-OA can sometimes be recovered steadily and slowly through avoidance measures.