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PubMed2006

[Qualitative and quantitative determination of the main components of huanglianjiedu decoction by HPLC-UV/MS].

AIM: To establish a comprehensive HPLC analytical method of Huanglianjiedu decoction. METHODS: This study was performed by HPLC-UV/MS to identify the chemical constituents of the whole and individual herbs of the "Huanglianjiedu decoction". Zorbax Extend C18 (150 mm x 4. 6 mm ID, 5 microm) column was used; the mobile phase was composed of acetonitrile (A) and water (B, with 0.5% acetic acid) with gradient elution; the flow rate was 1.0 mL x min(-1) and the column temperature was setup at 25 degrees C. The detection wavelength was 254 nm. RESULTS: The chromatogram of Huanglianjiedu decoction showed 21 main peaks. Peaks 1, 2, 5 and 18 were from Gardenia jasminoides Ellis, Peaks 8, 13, 14, 15, 16, 17, 19 and 21 from Scutellaria baicalensis Georgi. While 10 from Coptis chinensis Franch and 20 from Phellodendron amurense Rupr., Peaks 3, 4, 6, 9, 11 and 12 came from them together. Peak 7 presented in the chromatograms of the herbs except Gardenia jasminoides Ellis. By comparison of the retention time, the on-line UV spectra and MS spectra, 11 peaks were identified as 5 (geniposide), 9 (jatrorrhizine), 10 (coptisine), 11 (palmatine), 12 (berberine), 13 (baicalin), 15 (oroxin A), 17 (wogonoside), 19 (baicalein), 20 (obaculactone), 21 (wogonin), then eight of them were quantified by HPLC-UV. CONCLUSION: The method could represent the characteristics of Huanglianjiedu decoction, and it could be used to evaluate the quality and quantity of Huanglianjiedu decoction. It distinguished between Coptis chinensis Franch and Phellodendron amurense Rupr. by HPLC for the first time.

Berberine↗

PubMed2005

Screening and analysis of bioactive compounds with biofingerprinting chromatogram analysis of traditional Chinese medicines targeting DNA by microdialysis/HPLC.

Biofingerprinting chromatogram analysis, which is defined as the comparison of fingerprinting chromatograms of the extract of traditional Chinese medicines (TCMs) before and after the interaction with biological systems (DNA, protein, cell, etc.), was proposed for screening and analysis of the multiple bioactive compounds in TCMs. A method of microdialysis sampling combined with high performance liquid chromatography (HPLC) was applied to the study of DNA-binding property for the extracts of TCMs. Seven compounds were found to bind to calf thymus DNA (ct-DNA) from the TCMs of Coptis chinensis Franch (Coptis), but only three ones from Phellodendron amurense Rupr. (Phellodendron) and none from Sophoraflavescens Ait. (Sophora) to bind to ct-DNA, respectively. Three of them were identified as berberine, palmatine and jatrorrhizine and their association constants (K) to ct-DNA were determined by microdialysis/HPLC. Competitive binding behaviors of them to ct-DNA were also investigated.

Animals↗

PubMed2000

Identification of dyes on ancient Chinese paper samples using the subtracted shifted Raman spectroscopy method.

The Stein Collection in the British Library contains the Diamond Sutra, the world's oldest, dated, printed document. The paper of the Diamond Sutra and other documents from the Stein collection is believed to be dyed yellow by a natural extract, called huangbo, from the bark of Phellodendron amurense, which contains three major yellow chromophores: berberine, palmatine, and jatrorrhizine. Conservation of these documents requires definite information on the chemical composition of the dyes but no suitable, completely noninvasive analytical method is known. Here we report resonance Raman studies of a series of pure dyes, of plant materials and extracts, and of dyed ancient and modern paper samples. Resonance Raman spectroscopy is used to enhance the spectra of the dyes over the signals from the paper matrixes in which they are held. The samples all give resonance Raman spectra which are dominated by intense fluorescence, but by using SSRS (subtracted shifted Raman spectroscopy) we have obtained reliable spectra of the pure dyes, native bark from the Phellodendron amurense, modern paper dyed with huangbo extracted from this bark, and ancient paper samples. For both ancient paper samples whose pigment bands were detected, the relative intensities of the bands due to berberine and palmatine suggest that the ancient paper is richer in berberine than its modern counterpart. This is the first nondestructive in situ method for detection of these pigments in manuscripts, and as such has considerable potential benefit for the treatment of irreplaceable documents that are believed to be dyed with huangbo but documents on which conservation work cannot proceed without definite identification of the chemical compounds that they contain.

Luminescent Measurements↗

PubMed2002

Marked decrease of cyclosporin absorption caused by phellamurin in rats.

Phellamurin is a flavonoid glycoside that is abundant in the leaves of Phellodendron wilsonii Hayata et Kanehira (Rutaceae). In vitro everted rat intestine study indicated that phellamurin inhibited intestinal P-glycoprotein in a dose-dependent manner. In order to investigate the effect of phellamurin on cyclosporin absorption and disposition, rats were given cyclosporin (5 mg/kg) with or without phellamurin in a parallel design. Fluorescence polarization immunoassay was used to determine the blood concentration of cyclosporin. Unanticipatedly, our results indicated that the coadministration of phellamurin significantly decreased the Cmax of cyclosporin by 77 % and reduced the AUC(0-infinity) of cyclosporin by 56 %. This indicated that a serious interaction occurred between phellamurin with cyclosporin. To ensure the efficacy of cyclosporin, we suggest that the coadministration of phellamurin or Phellodendron wilsonii with cyclosporin should be avoided.

Animals↗

PubMed1988

The pharmacological and pathological studies on several hepatic protective crude drugs from Taiwan (I).

This study is to investigate the hepatic protective effect of several Taiwan crude drug extractions on the carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. The pharmacological and pathological effects of Bupleurum chinense, Phellodendron wilsonii, Clematis chinensis and Hedyotis corymbosa were analyzed by liver enzyme function test and pathological studies. However, the results of amine transferase SGOT and SGPT have shown a significant hepatic protective effect after treatment with Bupleurum chinense (P less than 0.005), Phellodendron wilsonii (P less than 0.001), Clematis chinensis (P less than 0.005) and Hedyotis corymbosa (P less than 0.005, SGPT only). The fatty degeneration around the central vein area and necrosis of the central lobule can be significantly improved by P. wilsonii and moderately changed by B. chinense or C. chinensis. Although fatty metamorphosis has been affected by H. corymbosa, various inflammatory cell infiltrations in the cytoplasm were noted.

Animals↗

PubMed2006

Assessing the potential of native tree species for carbon sequestration forestry in Northeast China.

Although the native forests of China are exceptionally diverse, only a small number of tree species have been widely utilized in forest plantations and reforestation efforts. We used dendrochronological sampling methods to assess the potential growth and carbon sequestration of native tree species in Jilin Province, Northeast China. Trees were sampled in and near the Changbaishan Biosphere Reserve, with samples encompassing old-growth, disturbed forest, and plantations. To approximate conditions for planted trees, sampling focused on trees with exposed crowns (dominant and co-dominant individuals). A log-linear relationship was found between diameter increment and tree diameter, with a linear decrease in increment with increasing local basal area; no significant differences in these patterns between plantations and natural stands were detected for two commonly planted species (Pinus koraiensis and Larix olgensis). A growth model that incorporates observed feedbacks with individual tree size and local basal area (in conjunction with allometric models for tree biomass), was used to project stand-level biomass increment. Predicted growth trajectories were then linked to the carbon process model InTEC to provide estimates of carbon sequestration potential. Results indicate substantial differences among species, and suggest that certain native hardwoods (in particular Fraxinus mandshurica and Phellodendron amurense), have high potential for use in carbon forestry applications. Increased use of native hardwoods in carbon forestry in China is likely to have additional benefits in terms of economic diversification and enhanced provision of "ecosystem services", including biodiversity protection.

Biodiversity↗

PubMed2004

Comparison of two Chinese medical herbs, Huangbai and Qianniuzi, on influence of short circuit current across the rat intestinal epithelia.

Huangbai (Phellodendron spec.) and Qianniuzi (Pharbitis spec.) are two traditional Chinese medical herbs used for anti-diarrheal and laxative agents, respectively. Ethanol and water extracts of these two herbs were prepared and effects of the extracts on ion transport of the rat intestinal epithelia were studied. For measuring changes of the short circuit current across the epithelia, the rat intestinal epithelia were mounted in the Ussing chamber and attached with voltage/current clamp. The intestinal epithelia were firstly activated by serosal administration of 5 microM forskolin. As current raised and being stable, extracts of these herbs were added, respectively, and changes in the short circuit current were recorded. Ethanol extract of Huangbai attenuated the current increment; on the contrary, ethanol extract of Qianniuzi augmented the current increment additionally. Water extracts of the two herbs showed minor effects on the current in comparison to ethanol extracts. The results provide evidences to reveal the pharmacological mechanism of the two Chinese medical herbs on the intestinal tissue.

Animals↗

PubMed2003

A new isocoumarin from bark of Pellodendron chinense.

A new isocoumarin, 3-acetyl-3,4-dihydro-5,6-dimethoxy-1H-2-benzopyran-1-one, was isolated from the cortex of Phellodendron chinense schneid. The structure was determined on the basis of spectroscopic evidences.

Coumarins↗

PubMed1999

[Comparison of semi-bionic extraction and water extraction in extraction of chemical constituents from processed cortex Phellodendri].

OBJECTIVE: To Study the granulation of processed Cortex Phellodendri. METHOD: Take berberine, total alkaloids and dry extract as indexes to compare the semi-bionic extraction (SBE) with water extraction (WE). RESULT: In 5 kinds of phellodendron bark solution, berberine, total alkaloids and dry extract obtained by SBE are much better than those obtained by WE. CONCLUSION: In the preparation of oral granules from processed Cortex Phellodendri, SBE is superior to WE, when the pH value is adjusted by HCl solution to 1.0 in the first decoction, adjusted by saturated Ca(OH)2 solution to 7.0 in the second decoction and to 10.0 in the third decoction respectively.

Alkaloids↗

PubMed2005

Acetaldehyde-induced interleukin-1beta and tumor necrosis factor-alpha production is inhibited by berberine through nuclear factor-kappaB signaling pathway in HepG2 cells.

Alcoholic liver disease (ALD) is one of the most common liver diseases in the world. Increased levels of proinflammatory cytokines, including interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), have been correlated with the patients affected by ALD. However, the direct effect of alcohol in the induction of IL-1beta and TNF-alpha has not been clarified. In this study, we demonstrated that acetaldehyde, the metabolic product of ethanol, was able to induce IL-1beta and TNF-alpha production in HepG2 cells. Nuclear factor-kappaB (NF-kappaB), the transcription factor involved in the regulation of cytokine production, was also activated by acetaldehyde through inhibitory kappaB-alpha (IkappaB-alpha) phosphorylation and degradation. However, the NF-kappaB inhibitors, such as aspirin, cyclosporin A and dexamethasone, inhibited both the acetaldehyde-induced NF-kappaB activity and the induced cytokine production. Therefore, these data suggested that acetaldehyde stimulated IL-1beta and TNF-alpha production via the regulation of NF-kappaB signaling pathway. By screening 297 controlled Chinese medicinal herbs supervised by Committee on Chinese Medicine and Pharmacy at Taiwan, we found that Coptis chinensis (Huang-Lien) and Phellodendron amurense (Huang-Po) were capable of inhibiting acetaldehyde-induced NF-kappaB activity. Berberine, the major ingredient of these herbs, abolished acetaldehyde-induced NF-kappaB activity and cytokine production in a dose-dependent manner. Moreover, its inhibitory ability was through the inhibition of induced IkappaB-alpha phosphorylation and degradation. In conclusion, we first linked the acetaldehyde-induced NF-kappaB activity to the induced proinflammatory cytokine production in HepG2 cells. Our findings also suggested the potential role of berberine in the treatment of ALD.

Acetaldehyde↗

PubMed2001

Topical anti-inflammatory activity of some Asian medicinal plants used in dermatological disorders.

The topical anti-inflammatory activity of extracts from Cassia angustifolia, Rheum palmatum, Coptis chinensis, Phellodendron amurense and Scutellaria baicalensis, plants used in traditional East Asian medicine against different skin disorders, was studied. Though in different degree, all the extracts significantly inhibited the edema induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), in both single or multiple application, oxazolone, and arachidonic acid (AA). None of the extracts inhibited in vitro the activity of phospholipase A(2) (PLA(2)) from Naja naja.

Administration, Topical↗

PubMed2002

Inhibition of Trypanosoma cruzi growth by medical plant extracts.

This study describes the screening of extracts obtained from 18 plants and two fungi used in the Chinese and Mediterranean traditional medicines on epimastigote forms of Trypanosoma cruzi. The extracts were tested against epimastigote of T. cruzi Bra C15C2 clone in vitro at 27 degrees C and at a concentration of 250 microg/ml in axenic culture. Angelica dahurica, A. pubescens, A. sinensis, Astragalus membranaceus, Coptis chinensis, Haplophyllum hispanicum, Phellodendron amurense, Poria cocos, Ranunculus sceleratus and Scutellaria baicalensis showed significant effects against the parasite with a percentage of growth inhibition between 20 and 100%. C. chinensis and R. sceleratus showed the greatest activity with IC(50) values of 1.7 microg/ml for C. chinensis and 10.7 microg/ml for R. sceleratus. These activities are greater than that of allopurinol. C. chinesis and R. sceleratus extracts did not show cytotoxic effects on rat polimorphonuclear cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and lactic dehydrogenase assays. These results allowed us to suggest that R. sceleratus and C. chinensis could be a source of new compounds clinically active against T. cruzi.

Animals↗

PubMed2003

Immunoquantitative analysis for berberine and its related compounds using monoclonal antibodies in herbal medicines.

The monoclonal antibody (MAb) against berberine, a bioactive constituent of Coptis japonica M., Phellodendron amurense R. and Hydrastis canadensis L., was produced and characterized. As immunogen, the derivative of berberine, 9-O-carboxymethyl berberrubine was synthesized and conjugated to carrier protein, bovine serum albumin (BSA). In order to confirm its immunogenicity, the ratio of hapten in berberine-BSA conjugate was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). After immunization, hybridomas secreting MAbs against berberine were produced by fusing splenocytes with mouse myeloma cell line, P3-X63-Ag8-653. After the screening, anti-berberine MAb 1D5-3B-7 was obtained. Subsequently, a quantitative ELISA system for berberine and its related compounds using the MAb was established and evaluated comparing with HPLC method. The ELISA method described in this study can be available as an analytical tool for quality control and standardization of medicinal plants and its prescriptions containing berberine and its related compounds.

Animals↗

PubMed1997

Inhibitory effects of oren-gedoku-to and its components on cholesteryl ester synthesis in cultured human hepatocyte HepG2 cells: evidence from the cultured HepG2 cells and in vitro assay of ACAT.

The pharmacological effects of Oren-gedoku-to (OGT), a Japanese-Chinese traditional herbal medicinal mixture on lipid biosynthesis were investigated in cultured human hepatocyte HepG2 cells. The addition of OGT (0.5 and 4.2 mg/ml), which had no effect on cell proliferation and cellular protein content, caused a marked decrease in the cellular cholesterol content, particularly cholesteryl ester content following 24 h incubation. The incorporation of 14C-oleate into cellular cholesteryl ester fraction was also reduced remarkably during incubation for 6 and 24 h. The effects of OGT, its components and its main active chemicals on acyl-coenzyme A:cholesterol acyltransferase (ACAT) activity were studied in vitro to explore the mechanism by which OGT inhibits cholesteryl ester formation. The data confirmed that OGT, in a dose-dependent manner, and its components (Scutellaria baicalensis, Coptis japonica, Gardenia jasminoides and Phellodendron amurense) remarkably inhibit ACAT activity. Among the main active chemicals of OGT, baicalein, a kind of flavonoid, decreased ACAT activity in a dose-dependent fashion from the level of 10(-6)M. These results strongly suggest that OGT reduces the cholesteryl ester formation in human hepatocytes by inhibiting ACAT, and that baicalein may, in part, be responsible for ACAT inhibition.

Carbon Radioisotopes↗

PubMed1993

A comparative study on commercial samples of phellodendri cortex.

A total of 31 commercial samples of Phellodendri cortex (Rutaceous plant) which originated from PHELLODENDRON AMURENSE Ruprecht, P. CHINENSE Schneid, P. WILSONII Hayata et Kanehira, and P. AMURENSE Rupr. var. SACHALINENSE Fr. Schm., respectively, were collected from the Taiwan and Japan herbal markets. The contents of five quaternary alkaloids (berberine, palmatine, jatrorrhizine, phellodendrine and magnoflorine) in these samples were determined by capillary electrophoresis. It was found that P. WILSONII (4.10 +/- 0.78%) and P. AMURENSE var. SACHALINENSE (4.18 +/- 1.03%) were superior to P. AMURENSE (1.55 +/- 0.72%) and P. CHINENSE (1.54 +/- 0.60%). Berberine was the major alkaloid in almost all samples. In comprised about 80% of the total alkaloids in the first two of the four herbs named above, but only about 40% in the latter two. From the data on the chemical analysis of the herb's constituents, as well as the herb's texture and color, we can postulate the origin and quality of a herb drug.

Journal Article↗

PubMed2005

Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) bacteria have been responsible for substantial morbidity and mortality in hospitals because they usually have multidrug resistance. Some natural products are candidates as new antibiotic substances. In the present study, we investigated the antimicrobial activity of berberine, the main antibacterial substance of Coptidis rhizoma (Coptis chinensis Franch) and Phellodendri cortex (Phellodendron amurense Ruprecht), against clinical isolates of MRSA, and the effects of berberine on the adhesion to MRSA and intracellular invasion into human gingival fibroblasts (HGFs). Berberine showed antimicrobial activity against all tested strains of MRSA. Minimum inhibition concentrations (MICs) of berberine against MRSA ranged from 32 to 128 microg/mL. Ninety percent inhibition of MRSA was obtained with 64 microg/mL or less of berberine. In the checkerboard dilution test, berberine markedly lowered the MICs of ampicillin and oxacillin against MRSA. An additive effect was found between berberine and ampicillin, and a synergistic effect was found between berberine and oxacillin against MRSA. In the presence of 1-50 microg/mL berberine, MRSA adhesion and intracellular invasion were notably decreased compared with the vehicle-treated control group. These results suggest that berberine may have antimicrobial activity and the potential to restore the effectiveness of beta-lactam antibiotics against MRSA, and inhibit the MRSA adhesion and intracellular invasion in HGFs.

Ampicillin↗

PubMed1999

Differential inhibitory effects of protoberberines on sterol and chitin biosyntheses in Candida albicans.

The anti-Candida potentials of 12 Korean medicinal plants were explored: methanol extracts from Coptis rhizoma and Phellodendron amurense caused significant inhibition of growth of Candida albicans, Candida glabrata, Candida krusei and Candida parapsilosis. The predominant active components of the extracts were the protoberberines berberine and palmatine; the most potent inhibition of growth was exhibited by berberine on C. krusei (MIC <4 mg/L) and palmatine on C. parapsilosis (MIC 16 mg/L). Both berberine and palmatine inhibited the in-vivo rate of incorporation of L-[methyl-14C]methionine into C-24 of ergosterol in C. albicans (50% inhibition concentration (IC50 values), 25 microM and 300 microM, respectively); this result suggests that sterol 24-methyl transferase (24-SMT) is one of the cellular targets for the antifungal activity of the protoberberines. In-vitro 24-SMT activity in microsomes from the yeast growth form of C. albicans was inhibited by both berberine (inhibition constant (Ki) 232 microM) and palmatine (Ki 257 microM) in a non-competitive manner; inhibition of 24-SMT was more marked for the mycelial form than for the yeast growth form of this organism. Palmatine inhibited chitin synthase from both the yeast and mycelial growth phases of C. albicans in a non-competitive manner (Ki 780 microM). The effects of protoberberines, extracted from established medicinal plants, on both sterol and cell wall biosyntheses in pathogenic fungi indicate that the potential of these compounds, or their semi-synthetic derivatives, as a novel class of antifungal agents should be investigated more fully.

Amphotericin B↗

PubMed2003

Suppressive effects of a Chinese herbal medicine qing-luo-yin extract on the angiogenesis of collagen-induced arthritis in rats.

Qing-Luo-Yin (QLY), a traditional Chinese herbal medicine for the treatment of rheumatoid arthritis, is a combination of the extracts of Sophora flavescens Ait., Phellodendron amurense Rupr., Sinomenium acutum Rehd. et Wils. and Dioscorea hypoglauca Palib. The suppressive effect of QLY on the development of angiogenesis was investigated in a rat model of collagen-induced arthritis (CIA). QLY (0.3 g/kg) was orally administered daily for 27 days. Neo-angiogenesis, pannus and cartilage damage, the expression of metalloproteinases (MMP)-3 messenger RNA (mRNA) and the level of the tissue inhibitor of matrix metalloproteinase (TIMP)-1 in the synovium were examined by histology, in situ hybridization and immunohistochemiscal assays, respectively. It was observed that the articular morphological alterations, the over-expression of MMP-3 mRNA and the reduced production of TIMP-1 in CIA rats were significantly ameliorated by QLY. QLY performed about as effectively as tripterygium glycosidorum tablets (0.1 g/kg) extracted from Tripterygium wilfordii Hook. f.. These results indicate that QLY exerts a suppressive effect on the angiogenesis of CIA rats, and suggest that the therapeutic effect of QLY could be due to restoring the balance of MMP-3 and TIMP-1 in rheumatoid synovium.

Animals↗