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Taxonomic and phytogeographic implications from ITS phylogeny in Berberis (Berberidaceae).

A phylogeny based on the internal transcribed spacer (ITS) sequences from 79 taxa representing much of the diversity of Berberis L. (four major groups and 22 sections) was constructed for the first time. The phylogeny was basically congruent with the previous classification schemes at higher taxonomic levels, such as groups and subgroups. A notable exception is the non-monophyly of the group Occidentales of compound-leaved Berberis (previously separated as Mahonia). At lower levels, however, most of previous sections and subsections were not evident especially in simple-leaved Berberis. Possible relationship between section Horridae (group Occidentales) and the simple-leaved Berberis clade implies paraphyly of the compound-leaved Berberis. A well-known South America-Old World (mainly Asia) disjunctive distribution pattern of the simple-leaved Berberis is explained by a vicariance event occurring in the Cretaceous period. The ITS phylogeny also suggests that a possible connection between the Asian and South American groups through the North American species ( Berberis canadensis or B. fendleri) is highly unlikely.

Asia↗

Possible mechanism of selective inotropic activity of the n-butanolic fraction from Berberis aristata fruit.

Berberis aristata is an edible plant employed in South Asian traditional medicine; in particular, its fruit is used as a tonic remedy for liver and heart. In isolated cardiac tissues, Berberis aristata fruit extract exhibits a positive inotropic action. Activity-directed fractionation using organic solvents revealed that the cardiotonic activity is concentrated in the n-butanolic fraction (BF). The cardiac action of BF was investigated in spontaneously beating right atria and in electrically driven right ventricular strips and left atria obtained from reserpinized guinea pigs. The results show that this fraction produces a dose-dependent positive inotropic action with little effect on heart rate. To study its possible mode of action, guinea pig atria were pretreated with propranolol, a beta-adrenoceptor blocking agent. This treatment abolished the cardiotonic effect of isoprenaline, whereas the cardiotonic effect of BF remained unaltered, suggesting that this effect does not involve stimulation of beta-adrenoceptors. On the other hand, application of carbachol reverses only part of the BF-induced increase in ventricular force of contraction, indicating that besides a cyclic AMP (cAMP)-dependent mechanism, a cAMP-independent mechanism underlies the inotropic action of BF. This is in line with the observation that the dynamics of isometric twitch contractions are not significantly altered by BF. Investigations in skinned myocardial preparations showed that BF modulates the calcium-dependent interaction of actin and myosin, apparently by reducing the cooperativity of the calcium-dependent binding of myosin to actin, i.e., there is enhanced calcium activation at low to physiological intracellular calcium, and reduced calcium activation at high intracellular calcium concentrations as present, for example, in ischemic calcium overload. These data indicate that the edible plant, Berberis aristata, contains active principle(s) that cause(s) a selective inotropic effect, involving-in the form of the modulatory effect on actin myosin cooperativity-a novel mechanism of action. Further phytochemical and pharmacological studies may lead to isolation and structural identification of an attractive, new cardiotonic agent from Berberis aristata fruit.

Adrenergic beta-Antagonists↗

[Determination of four alkaloids in Berberis plants by HPLC].

OBJECTIVE: In order to assess Berberis, a simple and effective high-performance liquid chromatographic (HPLC) method was established. METHOD: By applying HPLC with gradiation elution, this method was developed for determination of four kinds of alkaloids including berbamine, Jatrorrhizine, berberine and palmatine in methanolic extracts from root bark, root, stem bark and stem of Berberis. RESULT: The alkaloids berbamine, jatrorrhizine, berberine and palmatine shown good linear correlations in the range of 0.028-4.74 microg (r = 0.9998); 0.012-2.0 microg (r = 0.9996); 0.026-0.52 microg (r = 0.9999); 0.015-2.56 microg (r = 0.9998), respectively. The correlation coefficients of the calibration curve for the analytes exceeded 0.9998. CONCLUSION: The optimized HPLC method was applied to analyze various samples, and the results shown that the content of alkaloids were obvious difference in Berberis from different area, different species and different parts.

Alkaloids↗

Leaves of Berberidaceae (Berberis and Mahonia) from Oligocene sediments, near Tepexi de Rodríguez, Puebla.

From the Oligocene Los Ahuehuetes locality, near Tepexi de Rodríguez, Puebla, Mexico, five new plant species are described based on their leaf architecture. The presence of brochidodromous or acrodromous venation, and secondary veins forming angular (versus rounded) arcs, are well defined characters in the fossil material that relate it to Berberidaceae. Comparison with the leaves and leaflets of extant and fossil plants allow the recognition of one Mahonia and four Berberis new species. The lack of detailed information on leaf architecture in Berberidaceae limits the evaluation of the taxonomic relationships that can be suggested between fossil and extant plants. However, from a biogeographic point of view the presence of these new fossil plants supports the hypothesis of a North American origin of the Orientalis Groups of Mahonia, to which a lineage of Berberis may be added. Furthermore, two of the new species suggest the dispersal, some time during the Tertiary, of a lineage that today forms the Australis Group of Berberis from low latitude North America to South America. The movement of the Chortis Block is proposed as an alternative to explain the dispersal of a growing list of plants from north to south in the Americas. Only through future geological and palaeobotanical work can this hypothesis be corroborated.

Journal Article↗

Staphylococcus aureus MDR efflux pump inhibitors from a Berberis and a Mahonia (sensu strictu) species.

Bioactive fractionation, based on multi-drug resistance (MDR) pump inhibition in Staphylococcus aureus, resulted in the isolation of the active inhibitors 5'-methoxyhydnocarpin-D from leaves of Berberis (formerly Mahonia) trifoliolata and pheophorbide a from Berberis fendleri. The hydnocarpin derivative was not found in the latter species. Pheophytin a (the phytol derivative of pheophorbide a) was identified from both species, but it proved to have no MDR pump inhibitory activity. The somewhat uncommon, and inactive, flavonoid tricin was identified from B. trifoliolata. The occurrence of a flavonolignan in Mahonia-tpe species and its absence in Berberis sensu strictu may provide a chemical differentiation between the two groups which are now recombined on the basis of DNA studies. The strong bacterial efflux pump inhibition of pheophorbide a could be of importance as a plant defense against natural pathogens.

Journal Article↗

Estimation of heavy metals in different berberis species and its market samples.

Popularity of herbal drugs is increasing all over the world because of lesser side effects as compared to synthetic drugs. Besides, it costs less and easily available to poor people particularly in developing countries. But quality assurance of herbal drugs is very necessary prior to its use. Because in today's polluted environment, even herbal drugs are not safe. Berberis spp. is very important medicinal plant, having various medicinal properties. It is also included in Indian and British pharmacopoeias. Its demand is quite high in herbal drug market. So, to check the quality of market samples of this drug, ten different samples were procured from different drug markets of India for heavy metal estimation. Besides, genuine samples of four species of Berberis viz. B. aristata, B. chitria, B. lycium, B. asiatica were also collected from natural habitats to compare heavy metal concentration in both market and genuine samples. It was found that market samples were much more contaminated than genuine samples. Lead (Pb) concentration is far beyond from WHO permissible limit (10 ppm) for herbal drugs, reaching to maximum 49.75 ppm in Amritsar market sample. Likewise, concentration of all other metals like Cd, Cr and Ni were also very high in market samples as compared to genuine samples.

Berberis↗

A comparative study on the anti-inflammatory, antinociceptive and antipyretic effects of isoquinoline alkaloids from the roots of Turkish Berberis species.

Roots and barks of various Berberis species are used as folk remedy for the treatment of various inflammatory diseases such as lumbago, rheumatism and to reduce fever. Six isoquinoline alkaloids namely berberine, berbamine, palmatine, oxyacanthine, magnoflorine, and columbamine were isolated as the main components of alkaloidal fraction from the roots of Turkish Berberis species and effects were studied using various in vivo models in mice. All alkaloids inhibited inflammations in varying degrees, among them berberine, berbamine and palmatine were shown to possess significant and dose-dependent inhibitory activity against serotonin-induced hind paw oedema both on oral and topical applications and acetic acid-induced increase in vascular permeability on oral administration. Moreover, these three alkaloids were also shown to possess dose-dependent antinociceptive activity, which assessed by using the model based on the inhibition of p-benzoquinone-induced writhing movements as well as antipyretic activity on FCA-induced increased rectal temperature on subacute administration. However, all alkaloids induced gastric lesions in varying degrees.

Analgesics, Non-Narcotic↗

Berberis crataegina DC. root exhibits potent anti-inflammatory, analgesic and febrifuge effects in mice and rats.

Extracts obtained from the roots and barks of various Berberis species are used as folk remedy worldwide for the treatment of various inflammatory ailments including lumbago, rheumatism and to reduce fever. Effects of the extracts and fractions from the roots of Berberis crataegina DC. (Berberidaceae) were studied using various in vivo models of inflammation in mice and rats and observed potent inhibitory activity against carrageenan- and serotonin-induced hind paw oedema, acetic acid-induced increased vascular permeability, castor oil-induced diarrhoea, and Freund's complete adjuvant-induced (FCA) arthritis models. Through bioassay-guided fractionation berberine was isolated as the main active ingredient. Moreover, a dose-dependent analgesic activity was determined, which assessed by using the model based on the inhibition of acetic acid-induced writhing reflexes, as well as antipyretic activity on FCA-induced increased body temperature. Acute and subchronic toxicity studies were also performed.

Analgesics, Non-Narcotic↗

Berberis aetnensis C. Presl. extracts: antimicrobial properties and interaction with ciprofloxacin.

Previous research showed that berberine-containing Berberis species synthesise the substances 5'-methoxyhydnocarpin-D (5'-MHC-D) and pheophorbide a, which have no antimicrobial activity but inhibit the expression of multidrug resistant efflux pumps (MDRs) in Staphylococcus aureus and potentiate the action of berberine. The MDR pumps extrude synthetic and natural antimicrobials from bacterial cells. We searched for these compounds in Berberis aetnensis C. Presl. (Berberidaceae), an endemic plant of the volcano Mount Etna. This work confirms the presence of pheophorbide a and permits us to hypothesise the presence of 5'-MHC-D in leaf extracts. In fact, the activity of ciprofloxacin was improved when two chromatographic fractions isolated from leaf extracts were added. These results are indicative of the presence of MDR pump inhibitors. Moreover, crude extracts were tested on several micro-organisms and showed antimicrobial activity mainly against Gram-positive bacteria and yeasts.

Anti-Bacterial Agents↗

The antihypertensive and vasodilator effects of aqueous extract from Berberis vulgaris fruit on hypertensive rats.

The aqueous extract from Berberis vulgaris fruit (B.V.) was tested to evaluate its antihypertensive effects on DOCA-induced hypertension in the rats. Hypertension was induced in male Sprague-Dawley rats (200-250 g) by DOCA-salt injection (20 mg/kg, twice weekly, for 5 weeks, s.c.) plus NaCl (1%) which was added to the animals' drinking water. Then 5 weeks later, the rats were anaesthetized with thiopental (30 mg/kg, i.p.) and the arterial blood pressure was measured. The mean arterial blood pressure and heart rate were 231 +/- 6.4 (mmHg) and 506 +/- 12 (beats/min), respectively. Administration of B.V. extracts significantly reduced the rat arterial blood pressure. In in vitro studies, rings of descending aorta were cut and mounted for isometric tension recording in an organ chamber containing Krebs solution. Mesenteric beds were also removed and perfused with Krebs solution. After 1 h of stabilization, preparations (aortic rings or mesenteric beds) were precontracted with phenylephrine (10(-5) M), then different concentrations of B.V. (0.4, 2 and 4 mg/mL) were added which caused a relaxation in these vessels. To investigate the mechanism of action of the extract, the tissues were incubated with either L-NAME (10(-5) M) or indomethacin (10(-5) M) for 20 min. In the aortic rings L-NAME pretreatment could only reduce the vasodilatory effects of a low concentration of B.V. (0.4 mg/mL), but indomethacin was without effect. In isolated perfused mesenteric beds preincubation with either L-NAME or indomethacin did not modify the vasodilator effects of the aqueous extract from B.V. fruit. The present results suggest that the antihypertensive and vasodilatory effects of B.V. fruit extract are mainly endothelial-independent and it may be used to treat hypertension, a status with endothelial dysfunction.

Animals↗

Activity of Berberis aetnensis root extracts on Candida strains.

The antifungal activity of methanolic extract and alkaloidal fraction of Berberis aetnensis against Candida species was investigated. The crude extract was active against Candida species, this activity being higher than that of the alkaloidal fraction and berberine.

Antifungal Agents↗

A pharmacological study on Berberis vulgaris fruit extract.

Berberis vulgaris fruit (barberry) is known for its antiarrhythmic and sedative effects in Iranian traditional medicine. The effects of crude aqueous extract of barberry on rat arterial blood pressure and the contractile responses of isolated rat aortic rings and mesenteric bed to phenylephrine were investigated. We also examined effect of the extract on potassium currents recorded from cells in parabrachial nucleus and cerebellum rejoins of rat brain. Administration of the extract (0.05-1 mg/100 g body weight of rat) significantly reduced the mean arterial blood pressure and heart rate in anaesthetized normotensive and desoxycorticosteron acetate-induced hypertensive rats in a dose-dependent manner. Concentration-response curves for phenylephrine effects on isolated rat aortic rings and the isolated mesenteric beds in the presence of the extract were significantly shifted to the right. Application of the extract (1-50 microg/ml) shifted the activation threshold voltage to more negative potentials, leading to an enhancement in magnitude of the outward potassium current recorded from cells present in rat brain slices of parabrachial nucleus and cerebellum. This effect on potassium current may explain the sedative and neuroprotective effects of barberry. The present data support the hypothesis that the aqueous extract of barberry has beneficial effects on both cardiovascular and neural system suggesting a potential use for treatment of hypertension, tachycardia and some neuronal disorders, such as epilepsy and convulsion.

Animals↗

Study on the anti-inflammatory action of Berberis vulgaris root extract, alkaloid fractions and pure alkaloids.

Extracts obtained from the roots of Berberidaceae species have been used in Eastern and Bulgarian folk medicine in rheumatic and other chronic inflammatory disorders. The investigations of the chemical composition and immunological properties show that their activity is mainly due to the alkaloid constituents. In the present study the anti-inflammatory properties of total ethanol extract (TEE), three alkaloid fractions, a major alkaloid berberine and oxyacanthine isolated from Berberis vulgaris roots were compared. All these were applied in acute inflammation (carrageenan- and zymosan-induced paw oedema), as the TEE showed the highest reducing effect. Their ability to alter in vivo and in vitro complement activity was determined. Also, the TEE was most effective in a chronic inflammatory model of adjuvant arthritis. The protoberberine fractions Bv2, Bv3 and berberine suppressed a delayed type hypersensitivity (DTH) reaction. Fraction Bv1 and berberine diminished antibody response against SRBC in vivo. The in vitro treatment of splenocytes with berberine showed that the anti-SRBC antibody synthesis was influenced in a different manner depending on the time course of its application. Oxyacanthine was less effective than berberine in the tests used.

Alkaloids↗

Antihistaminic and anticholinergic activity of barberry fruit (Berberis vulgaris) in the guinea-pig ileum.

Barberry (Berberis vulgaris) is a well known medicinal plant in Iran and has also been used as food. The antihistaminic and anticholinergic activity of aqueous extract of barberry fruits were investigated on isolated guinea-pig ileum, and dose response curves of histamine and acetylcholine with and without extract were plotted. The pA2 values for antihistaminic activity of extract and dexchlorpheniramine were calculated (extract; pA2 +/- S.E.M. = 4.50 +/- 0.01[-log C (g/l)]; dexchlorpheniramine; pA2 +/- S.E.M. = 9.36 +/- 0.14[-log C (M)]) and compared with each other. The pA2 values for anticholinergic activity of extract and atropine were also calculated (extract, pA2 +/- S.E.M. = 4.37 +/- 0.03[-log C (g/l)]; atropine, pA2 +/- S.E.M. = 8.99 +/- 0.13[-log C (M)]) and compared. The results indicated antihistaminic and anticholinergic activity of extract that seems to be of the competitive type.

Animals↗

5'-Methoxyhydnocarpin-D and pheophorbide A: Berberis species components that potentiate berberine growth inhibition of resistant Staphylococcus aureus.

A new method of bioactivity-directed fractionation, based on multidrug resistant pump (MDR) inhibition in Staphylococcus aureus, was demonstrated. This resulted in the isolation, from berberine-containing Berberis species, of two compounds that are themselves devoid of S. aureus antibacterial activity, but that form potent synergistic couples with a subinhibitory concentration of berberine. The bacterial MDR pump inhibitors were identified as the flavonolignan 2 and the porphyrin 3. Another natural flavonolignan, silybin (8) from Silybum marianum, was also shown to be a bacterial MDR pump inhibitor.

Berberine↗

Molecular cloning and heterologous expression of a cDNA encoding berbamunine synthase, a C--O phenol-coupling cytochrome P450 from the higher plant Berberis stolonifera.

A cDNA encoding a cytochrome P450-dependent oxidase, berbamunine synthase (EC 1.1.3.34; CYP80), from cell suspension cultures of the higher plant Berberis stolonifera Koehne and Wolf (barberry) has been isolated and heterologously expressed in functional form in insect cell culture using a baculovirus-based expression system. This cytochrome P450-dependent enzyme is unusual in that it catalyzes the regio- and stereoselective formation of a C--O phenol couple in bisbenzylisoquinoline alkaloid biosynthesis without concomitant incorporation of activated oxygen into the product. Consistent with the function of an oxidase rather than a monooxygenase, an essential glycine residue in the distal helix, which forms the oxygen-binding pocket in the well-studied bacterial enzyme P-450cam, is replaced by proline at the equivalent position in berbamunine synthase. This oxidase was accumulated in an active form in insect cell microsomes and accepted electrons from the endogenous NADPH-cytochrome P450 reductase. The heterologously expressed enzyme oxidatively couples either two molecules of (R)-N-methylcoclaurine to form the (R,R) dimer guattegaumerine or one molecule each of (R)- and (S)-N-methylcoclaurine to form the (R,S) dimer berbamunine. The ratio of the two bisbenzylisoquinolines formed could be altered by reductase source or by varying the enantiomer composition of the substrates.

Alcohol Oxidoreductases↗

Purification and properties of (S)-tetrahydroprotoberberine oxidase from suspension-cultured cells of Berberis wilsoniae.

A novel oxidase, catalyzing in the presence of oxygen the removal of four hydrogen atoms from a number of tetrahydroprotoberberines with simultaneous production of 1 mol H2O2 and H2O each, has been discovered and purified to homogeneity from Berberis wilsoniae cell cultures. This enzyme, (S)-tetrahydroprotoberberine oxidase, exhibited strict specificity for the (S)-enantiomer of tetrahydroprotoberberines and 1-benzylisoquinoline alkaloids, a pH optimum at 8.9, a molecular mass of 105 kDa and consisted of two subunits each of 53 kDa and covalently bound flavin. The Km values for (S)-scoulerine and (S)-norreticuline were 25 microM and 150 microM respectively. Concentration of the end-products, either protoberberines or H2O2, greater than 0.5 mM caused severe enzyme inhibition. This catalyst was responsible for the conversion of (S)-tetrahydrocolumbamine to the key intermediate, columbamine, in the metabolic pathway leading to berberine, jatrorrhizine and palmatine.

Berberine Alkaloids↗