Hepatoprotective effect of shidagonglao on acute liver injury induced by carbon tetrachloride.

This study investigates the hepatoprotective activity of ethanol extract from Shidagonglao roots (SDGL(EtOH)). The hepatoprotective effect of SDGL(EtOH) (20, 100 and 500 mg/kg) was analyzed on carbon tetrachloride (CCl(4))-induced acute liver injury. Rats pretreated orally with SDGL(EtOH) (100 and 500 mg/kg) and silymarin (200 mg/kg) for 3 consecutive days prior to the administration of a single dose of 50% CCl(4) (0.10 ml/100 g of bw, ip) significantly prevented the increases in the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in CCl(4)-treated rats. Histological analysis also showed that SDGL(EtOH) (100 and 500 mg/kg) and silymarin reduced the incidence of liver lesions including vacuole formation, neutrophil infiltration and necrosis of hepatocytes induced by CCl(4) in rats. Moreover, the SDGL(EtOH) (100 and 500 mg/kg) increased the activities of anti-oxidative enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GRd) and decreased malondialdehyde (MDA) level in liver, as compared to those in the CCl(4)-treated group. Furthermore, SDGL(EtOH) (100 and 500 mg/kg) and silymarin attenuated the increased levels of tumor necrosis factor-alpha (TNF-alpha) in serum and nitric oxide (NO) in liver as compared to the CCl(4)-treated group. The hepatoprotective mechanisms of SDGL(EtOH) are likely related to inhibition of TNF-alpha, MDA and NO productions via increasing the activities of antioxidant enzymes (SOD, GPx and GRd). These experimental results suggest that SDGL(EtOH) can attenuate CCl(4)-induced acute liver injury in rats.

Analgesic and anti-inflammatory activities of ethanol root extract of Mahonia oiwakensis in mice.

AIMS OF THE STUDY: This study investigated the anti-inflammatory and analgesic activities, and protoberberine alkaloid contents of ethanol extract of MO roots (MOR(EtOH)). MATERIALS AND METHODS: The analgesic activity of MOR(EtOH) was determined using acetic acid-induced writhing response and formalin test. The anti-inflammatory activity of MOR(EtOH) was determined using the lambda-carrageenan-induced paw oedema model. The protoberberine alkaloid contents of MOR(EtOH) were identified by high-performance liquid chromatography (HPLC). RESULTS: MOR(EtOH) (100 and 500 mg/kg) decreased the acetic acid-induced writhing responses and licking times of the second phase in the formalin test. Moreover, carrageenan-induced paw oedema was significantly reduced in a dose-dependent manner by administering MOR(EtOH) (100 and 500 mg/kg) at 3, 4, and 5h after the carrageenan injection. The serum levels of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) of MOR(EtOH)-treated mice were significantly reduced compared with those in the serum of animals administered carrageenan. Notably, MOR(EtOH) attenuated the expression of cyclo-oxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) and neutrophil infiltration in paw tissues injected with carrageenan. The anti-inflammatory mechanisms of MOR(EtOH) appear to be related to the inhibition of neutrophil infiltration, iNOS and COX-2 protein expression, NO release, and the decreasing TNF-alpha level in serum. The analytical results showed that the contents of berberine, palmatine and jatrorrhizine were 191.45 mg/g extract, 100.15 mg/g extract and 66.45 mg/g extract, respectively. CONCLUSION: These experimental results suggest that MOR(EtOH) produced both analgesic and anti-inflammatory effects in mice and may be a candidate for the development of pharmacological agents used in the treatment of inflammatory disorders.

Oxidative metabolism of the alkaloid rutaecarpine by human cytochrome P450.

Rutaecarpine is the main active alkaloid of the herbal medicine, Evodia rutaecarpa. To identify the major human cytochrome P450 (P450) participating in rutaecarpine oxidative metabolism, human liver microsomes and bacteria-expressed recombinant human P450 were studied. In liver microsomes, rutaecarpine was oxidized to 10-, 11-, 12-, and 3-hydroxyrutaecarpine. Microsomal 10- and 3-hydroxylation activities were strongly inhibited by ketoconazole. The 11- and 12-hydroxylation activities were inhibited by alpha-naphthoflavone, quinidine, and ketoconazole. These results indicated that multiple hepatic P450s including CYP1A2, CYP2D6, and CYP3A4 participate in rutaecarpine hydroxylations. Among recombinant P450s, CYP1A1 had the highest rutaecarpine hydroxylation activity. Decreased metabolite formation at high substrate concentration indicated that there was substrate inhibition of CYP1A1- and CYP1A2-catalyzed hydroxylations. CYP1A1-catalyzed rutaecarpine hydroxylations had V(max) values of 1,388 to approximately 1,893 pmol/min/nmol P450, K(m) values of 4.1 to approximately 9.5 microM, and K(i) values of 45 to approximately 103 microM. These results indicated that more than one molecule of rutaecarpine is accessible to the CYP1A active site. The major metabolite 10-hydroxyrutaecarpine decreased CYP1A1, CYP1A2, and CYP1B1 activities with respective IC(50) values of 2.56 +/- 0.04, 2.57 +/- 0.11, and 0.09 +/- 0.01 microM, suggesting that product inhibition might occur during rutaecarpine hydroxylation. The metabolite profile and kinetic properties of rutaecarpine hydroxylation by human P450s provide important information relevant to the clinical application of rutaecarpine and E. rutaecarpa.

Identification of the microsomal oxidation metabolites of rutaecarpine, a main active alkaloid of the medicinal herb Evodia rutaecarpa.

Rutaecarpine is a quinazolinocarboline alkaloid of the medicinal herb Evodia rutaecarpa and shows a variety of pharmacological effects. Four oxidation metabolites of rutaecarpine were prepared from 3-methylcholanthrene-treated rat liver microsomes. These metabolites had an [M + H]+ ion at m/z 304. The structures of metabolites were identified by comparison of their liquid chromatograms and mass, absorbance, and 1H NMR spectra with those of synthetic standards. Rutaecarpine was metabolized by microsomal enzymes to form 3-, 10-, 11-, and 12-hydroxyrutaecarpine. The formation of 10-hydroxyrutaecarpine was highly induced by a cytochrome P450 1A inducer, 3-methylcholanthrene.

Analysis of synthetic anti-diabetic drugs in adulterated traditional Chinese medicines by high-performance capillary electrophoresis.

Four synthetic anti-diabetic drugs, acetohexamide (ACE), chlorpropamide (CHL), glibenclamide (GLI) and tolbutamide (TOL), which can be found as adulterants in traditional Chinese medicines (TCMs) were assayed simultaneously using high-performance capillary electrophoresis (HPCE) in 4 min with UV detection at 200 nm. The electrolyte was a buffer solution containing 100 mM phosphate buffer (NaH2PO4/Na2B4O7, pH 7.5). Applied voltage was 15.0 kV and temperature was 30 degrees C. 2-(4-Hydroxyphenyl) ethyl ammonium chloride (HEA) was used as an internal standard. The effects of buffer concentration, pH and supplied voltage on separation were investigated. The relative standard deviations (R.S.D.) of these anti-diabetic drugs for intra-day and inter-day analyses were 0.23-4.27 and 1.23-6.33%, respectively. The recoveries of the synthetic drug adulterants in traditional Chinese medicinal formula ranged from 81.3 to 105.5%. GLI was found and determined in a real sample of TCM.

Determination of matrine and oxymatrine in Sophora subprostata by CE.

Shan-dou-gen is the dried roots of Sophora subprostata (Leguminosae) and a commonly used Chinese herbal drug in Taiwan. It possesses antipyretic, anti-inflammatory, analgesic effects and is used to treat sore throat and acute pharyngolaryngeal infections. To evaluate the quality of S. subprostata, a simple, rapid and accurate high-performance capillary electrophoresis (HPCE) method was developed for the assay of two alkaloids: matrine and oxymatrine. The electrolyte was a buffer solution containing 75% 130 mM phosphate buffer (NaH2PO4/H3PO4, pH 3.5) and 25% acetonitrile. Applied voltage was 10 kV and temperature was 30 degrees C. 2-(4-Hydroxyphenyl)ethylammonium chloride was used as an internal standard and detector set at 200 nm. The contents of matrine and oxymatrine of S. subprostata in several different samples of crude drugs and commercial concentrated preparation have also been determined.

Preventive effect of the Taiwan folk medicine Ixeris laevigata var. oldhami on alpha-naphthyl-isothiocyanate and carbon tetrachloride-induced acute liver injury in rats.

The hepatoprotective effects of Ixeris laevigata Sch-Bip. var. oldhami Kitam. (IL) were studied on cholestatic hepatitis induced by alpha-naphthylisothiocyanate (ANIT, 100 mg/10 mL/kg, in olive oil, i.p.) and acute hepatitis induced by carbon tetrachloride (20% CCl(4)/olive oil, 1.5 mL/kg, i.p.) in rats. Hepatoprotective activity was monitored by estimating the serum transaminases levels and the histopathological changes in the livers of experimental rats. The pretreatment of animals with IL, extract (0.3-2.0 g/kg orally) significantly inhibited the acute elevation of serum transaminases, as well as the hepatotoxin-induced histopathological changes in the livers of the experimental rats.