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Boldo and boldine: an emerging case of natural drug development.
Speisky H, Cassels BK.

Unidad de Bioquimica Farmacologica y Lipidos, Universidad de Chile, Santiago.

Boldo (Peumus boldus Mol.), a Chilean tree traditionally employed in folk medicine and recognized as a herbal remedy in a number of pharmacopoeias, mainly for the treatment of liver ailments, has recently been the subject of increasing attention. Boldine, in particular, the major and most characteristic alkaloidal constituent of this plant species, now emerges as its most interesting active principle from the pharmacological viewpoint. The recent demonstration that boldine is an effective antioxidant in both biological and non-biological systems has opened up the perspective of a broad range of uses in medicine and industry. Given the toxicological data on this alkaloid, its antioxidative properties situate it as a potentially useful substance in many disease states featuring free-radical related oxidative injury. This review attempts to cover and discuss the studies conducted over the last four decades on the chemical and pharmacological properties of boldo and its main constituent.

PMID: 8202440 [PubMed - indexed for MEDLINE]



Composition and antimicrobial activity of the essential oil of Peumus boldus leaves.
Vila R, Valenzuela L, Bello H, Montes M, Adzet T.

The composition and the antimicrobial activity of the essential oil from the leaves of Peumus boldus is investigated. Analyses of the oil obtained by hydrodistillation were carried out by GC and GC-MS using columns of two different stationary phases. Fractionation of the essential oil by column chromatography on silica gel was performed to improve identification of some constituents. More than 90% of the total oil (46 components) was identified, major constituents being monoterpenes (90.5%), among which limonene (17.0%), p-cymene (13.6%), 1.8-cineole (11.8%), and beta-phellandrene (8.4%) reached the highest percentages. Determination of the minimal bactericidal or fungicidal concentration against several microorganisms showed interesting activities towards Streptococcus pyogenes, Micrococcus sp., and Candida sp.

PMID: 10193210 [PubMed - indexed for MEDLINE]



Protective effects of boldine against free radical-induced erythrocyte lysis.
Garrido A, Bannach R, Gotteland M, Speisky H.

Laboratory of Lipids and Antioxidants, Nutrition and Food Technology Institute, University of Chile, Santiago.

Boldine, an aporphine alkaloid extracted from the leaves and bark of boldo (Peumus boldus Mol.), has been shown to exhibit strong free-radical scavenger and antioxidant properties. Here, we report the in vitro ability of boldine to protect intact red cells against the haemolytic damage induced by the free radical initiator 2, 2'-azobis-(2-amidinopropane) (AAPH). Boldine concentration-dependently prevented the AAPH-induced leakage of haemoglobin into the extracellular medium. Substantial and similar cyto-protective effects of boldine were observed whether the antioxidant was added 1 h prior to, or simultaneously with, the azo-compound. The delayed addition of boldine, by 1 h relative to AAPH, diminished but did not abolish its cytoprotective effect. However, negligible effects of boldine were observed after its addition to erythrocytes previously incubated with AAPH for 2 h. The data presented demonstrate that, in addition to its well-established antioxidant effects, boldine also displays time-dependently strong cytoprotective properties against chemically induced haemolytic damage. Copyright 2000 John Wiley & Sons, Ltd.

PMID: 10925398 [PubMed - indexed for MEDLINE]



[Effect of a dry boldo extract on oro-cecal intestinal transit in healthy volunteers]
Gotteland M, Espinoza J, Cassels B, Speisky H.

Unidad de Gastroenterologá, INTA, Santiago de Chile.

BACKGROUND: Boldo (Peumus boldus Molina) is a widely used medicinal plant. However, its physiological effects are not well known. Recent studies in animals showed that certain components of boldo relax smooth muscle and prolong intestinal transit. AIM: To assess the effects of a dry boldo extract on oro cecal transit time in normal humans. SUBJECTS AND METHODS: Twelve volunteers received 2.5 g of a dry boldo extract or a placebo (glucose) during two successive periods of four days. On the fourth day, 20 g of lactulose were administered and breath hydrogen was collected every 15 min. Oro cecal transit time was defined as the time in which breath hydrogen increased by 20 ppm over the fasting level. RESULTS: Oro cecal transit time was larger after dry boldo extract administration, compared to placebo (112.5 +/- 15.4 and 87 +/- 11.8 min respectively, paired t p < 0.05). CONCLUSIONS: Dry boldo extract prolongs oro cecal transit time, a possible explanation for its medicinal use

PMID: 8657963 [PubMed - indexed for MEDLINE]



Protective effect of boldine on oxidative mitochondrial damage in streptozotocin-induced diabetic rats.
Jang YY, Song JH, Shin YK, Han ES, Lee CS.

Department of Pharmacology, College of Medicine, Chung-Ang University, Seoul, 156-756, Korea.

Increased oxidative stress has been suggested to be involved in the pathogenesis and progression of diabetic tissue damage. Several antioxidants have been described as beneficial for oxidative stress-associated diseases. Boldine ([s]-2,9-dihydroxy-1, 10-dimethoxyaporphine) is a major alkaloid found in the leaves and bark of boldo (Peumus boldus Molina), and has been shown to possess antioxidant activity and anti-inflammatory effects. From this point of view, the possible anti-diabetic effect of boldine and its mechanism were evaluated. The experiments were performed on male rats divided into four groups: control, boldine (100 mg kg(-1), daily in drinking water), diabetic [single dose of 80 mg kg(-1)of streptozotocin (STZ), i.p.] and diabetic simultaneously fed with boldine for 8 weeks. Diabetic status was evaluated periodically with changes of plasma glucose levels and body weight in rats. The effect of boldine on the STZ-induced diabetic rats was examined with the formation of malondialdehydes and carbonyls and the activities of endogenous antioxidant enzymes (superoxide dismutase and glutathione peroxidase) in mitochondria of the pancreas, kidney and liver. The scavenging action of boldine on oxygen free radicals and the effect on mitochondrial free-radical production were also investigated. The treatment of boldine attenuated the development of hyperglycemia and weight loss induced by STZ injection in rats. The levels of malondialdehyde (MDA) and carbonyls in liver, kidney and pancreas mitochondria were significantly increased in STZ-treated rats and decreased after boldine administration. The activities of mitochondrial manganese superoxide dismutase (MnSOD) in the liver, pancreas and kidney were significantly elevated in STZ-treated rats. Boldine administration decreased STZ-induced elevation of MnSOD activity in kidney and pancreas mitochondria, but not in liver mitochondria. In the STZ-treated group, glutathione peroxidase activities decreased in liver mitochondria, and were elevated in pancreas and kidney mitochondria. The boldine treatment restored the altered enzyme activities in the liver and pancreas, but not the kidney. Boldine attenuated both STZ- and iron plus ascorbate-induced MDA and carbonyl formation and thiol oxidation in the pancreas homogenates. Boldine decomposed superoxide anions, hydrogen peroxides and hydroxyl radicals in a dose-dependent manner. The alkaloid significantly attenuated the production of superoxide anions, hydrogen peroxide and nitric oxide caused by liver mitochondria. The results indicate that boldine may exert an inhibitory effect on STZ-induced oxidative tissue damage and altered antioxidant enzyme activity by the decomposition of reactive oxygen species and inhibition of nitric oxide production and by the reduction of the peroxidation-induced product formation. Boldine may attenuate the development of STZ-induced diabetes in rats and interfere with the role of oxidative stress, one of the pathogeneses of diabetes mellitus.

PMID: 10987997 [PubMed - indexed for MEDLINE]



Botanical perspectives on health peppermint: more than just an after-dinner mint.
Spirling LI, Daniels IR.

Epsom General Hospital, Surrey, England.

Throughout history different species of mint have been used across the globe for their varying properties, both medicinal and culinary. Today, the commercial sales of mints are expanding each year--and at the end of a large meal after-dinner mints are frequently served. But why do we take them? Peppermint (Mentha piperita) is usually taken after a meal for its ability to reduce indigestion and colonic spasms by reducing the gastrocolic reflex. It is a naturally occurring hybrid cross between water mint (M. aquatica) and spearmint (M. spicata) and is best known for its role as a popular flavouring agent. Less well recognised is peppermint's potential role in the management of numerous other medical conditions including certain procedures, e.g. colonoscopy. With the growing popularity of herbal remedies, among both the public and medical practitioners, it would seem that now is an opportune time to consider further what peppermint has to offer the world of medicine.

PMID: 11329700 [PubMed - indexed for MEDLINE]



Influence of dietary supplementation of herb extracts on volatile sulfur production in pig large intestine.
Ushid K, Maekawa M, Arakawa T.

Laboratory of Animal Science, Kyoto Prefectural University, Shimogamo, Kyoto, Japan.

Volatile sulfur compounds (VS) are generated in the large intestine by the bacterial metabolism of sulfate and sulfur amino acids. VS are potentially harmful to the host. The effect of dietary supplementation of herb extracts on volatile sulfur production in the large intestine of pig was evaluated in this study. The extracts Perilla frutescens (Soyou), Mentha piperita (Peppermint), and Ajuga decumbens (Kiransou) were fed to pigs equipped with a permanent cannula at the cecum. Cecal digesta were sampled and analyzed for ammonia and short-chain fatty acids (SCFA). Sampled digesta were incubated anaerobically either with or without L-methionine for 24 h to estimate volatile sulfur production in vivo. L-Methionine was supplemented to enhance methanethiol (MeSH) production. At the end of the incubation, head space concentrations of volatile sulfur compounds such as hydrogen sulfide (H2S), MeSH, and dimethyl sulfide (DMS) were determined by flame-photometric gaschromatography after the addition of 6 N HCl. Sampled digesta were also subjected to the most probable number estimations for sulfate-reducing bacteria (SRB), sulfide producer from L-methionine, and MeSH producers from L-methionine. All three herb extracts significantly decreased H2S (p<0.05), MeSH (p<0.05), and ammonia (p<0.05) production, but SCFA production was not affected (p>0.05). The number of volatile sulfur-producing bacteria did not vary among groups by the dietary supplementation of these herb extracts. Serial solvent extraction was done on these herb extracts to specify the active fractions that reduce volatile sulfur production. n-Butanol fraction of all three extracts significantly reduced volatile sulfur production in vitro.

PMID: 12026183 [PubMed - indexed for MEDLINE]



Antimicrobial screening of Mentha piperita essential oils.
Kirimer N, Demirci F.

Medicinal and Aromatic Plant and Drug Research Centre (TBAM), Anadolu University, 26470-EskiÅŸehir, Turkey.

Essential oils of peppermint Mentha piperita L. (Lamiaceae), which are used in flavors, fragrances, and pharmaceuticals, were investigated for their antimicrobial properties against 21 human and plant pathogenic microorganisms. The bioactivity of the oils menthol and menthone was compared using the combination of in vitro techniques such as microdilution, agar diffusion, and bioautography. It was shown that all of the peppermint oils screened strongly inhibited plant pathogenic microorganisms, whereas human pathogens were only moderately inhibited. Chemical compositions of the oils were analyzed by GC and GC/MS. Using the bioautography assay, menthol was found to be responsible for the antimicrobial activity of these oils.

PMID: 12083863 [PubMed - indexed for MEDLINE]



In vivo and in vitro effects of bromelain on PGE2 and SP concentrations in the inflammatory exudate in rats
GASPANI Leda ; LIMIROLI Elena ; FERRARIO Paolo ; BIANCHI Mauro ;

Abstract
The effects of bromelain were examined in rats with subcutaneous carrageenin-induced inflammation. After oral in vivo administration, bromelain (10 and 20 mg/kg p.o.) induced a significant decrease of both PGE2 and substance P concentrations in the exudate. When added to the inflammatory exudate in vitro, the drug (25, 50, 100 µg/ml) did not affect PGE2 concentrations and induced an increase in the substance P levels. Our data indicate that bromelain reduces the production of two key mediators of inflammation. This effect does not seem to be related to a direct action of the drug on PGE2 and SP released in the exudate in response to the inflammatory stimulus.

Pharmacology ISSN 0031-7012 CODEN PHMGBN



Inhibitory effect of enzyme therapy and combination therapy with cyclosporin A on collagen-induced arthritis.
Rovenská E, Svík K, Stancíkova M, Rovenský J

Research Institute of Rheumatic Diseases, Nábrezie I. Krasku 4, 921 01 Piest'any, Slovak Republic. rovensky@vurch.sk

OBJECTIVE: There is increasing interest in the use of combination therapy for rheumatoid arthritis and in the possibility of combining the conventional drug approach with newer antirheumatic therapy. The present study investigates the efficacy of long-term prophylactic enzyme therapy and combination therapy with cyclosporin A in rats with collagen-induced arthritis. METHODS: Rats with collagen-induced arthritis were administered the following drugs: cyclosporin A (5 mg/kg/day and 10 mg/kg/day orally); a mixture of enzymes containing pure substances (bromelain, trypsin, rutin) in the same ratio as in Phlogenzym (PHL, 150 mg/kg, twice daily intrarectally); and a combination of 5 mg/kg/day cyclosporin A plus 300 mg/kg/day PHL for a period of 50 days from the immunization. Levels of serum albumin, serum nitrite/nitrate concentrations, changes in hind paw swelling and bone erosions were measured in the rats as variables of inflammation and destructive arthritis-associated changes. RESULTS: Treatment with 10 mg/kg cyclosporin A, as well as combination therapy with half dosages of cyclosporin A (5 mg/kg) plus PHL significantly inhibited both inflammation and destructive arthritis-associated changes. Significant differences in favor of combination therapy with 5 mg/kg CsA + 300 mg/kg PHL as compared to 5 mg/kg CsA alone were seen in hind paw swelling. Also, reduction of the radiographic scores was more significant in the combination therapy group. Five mg cyclosporin A or PHL alone reduced the disease markers studied to a lesser extent, and in the case of enzyme therapy this occurred at a later stage of arthritis development. CONCLUSION: Our results show the inhibitory effect of enzyme therapy on collagen-induced arthritis in rats, as well as the efficacy of cyclosporin A given in low doses in combination with enzyme therapy, which may be useful in the treatment of rheumatoid arthritis.

PMID: 11407084 [PubMed - indexed for MEDLINE]



Effect of bromelain on kaolin-induced inflammation in rats.
Kumakura S, Yamashita M, Tsurufuji S.

Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.

The effects of stem bromelain on the plasma kallikrein system, bradykinin levels and plasma exudation at the inflammatory site were examined in rats with a kaolin-induced inflammation of an air pouch. Bromelain (5, 7.5 mg/kg) caused a dose-dependent decrease of bradykinin levels at the inflammatory site and a parallel decrease of the prekallikrein levels in sera. Plasma exudation was also reduced dose dependently. Bradykinin-degrading activity in sera was elevated after treatment with bromelain, although it was unchanged in the pouch fluid. These data indicate that bromelain inhibits plasma exudation through inhibiting the generation of bradykinin at the inflammatory site via depletion of the plasma kallikrein system.

PMID: 3046953 [PubMed - indexed for MEDLINE]



Bromelain Proteases Reduce Human Platelet Aggregation in Vitro, Adhesion to Bovine Endothelial Cells and Thrombus Formation in Rat Vessels in Vivo
CAROLA METZIG, EDYTA GRABOWSKA, KLAUS ECKERT, KLAUS REHSE and H. RAINER MAURER

1 Harrach T, Eekcrt K, Schulze-Forster K, Nuck R, Grunow D und Maurer HR: Isolation and partial characterization of basic proteiiiases from stem bromelain. J Protein Chem 14: 41-52.
2 Harrach T, Eckert K, Maurer HR, Machleidt I, Machleidt W and Nuck R: Isointion and characterization of two forms of an acidic bromelain stem proteinasc. J Protein Chem.
3 Taussig SJ and Bntkin S: Bromelain, the enzyme complex of pineapple (Ananas comosus) and its clinical application. An update. J Ethnopharm 22: 191-203.

Abstract.
The thiol protease, bromelain, an extract from pine apple stern, was suggested to have antuhrombotic and anticoagulant activities in vivo. We studied the effects of bromelain on cell size distribution of isolated human platelets in vitro by Coulter Counter measurements. Preincubation of platelets 'with bromelain (10 ug/mL) completely prevented the thrombin (0.2 U/mL) induced platelet aggregation. Papain was less active in pi-eventing platelet aggregation. In vitro, bromelain (0.1 ug/mL) reduced the adhesion of bound, thrombin stimulated, fluorescent labeled platelets to bovine aorta endolhelial cells. In addition, preincubation of platelets with bromelain, prior to thrombin activation, reduced the platelet adhesion to the endothelial cells to the low binding value of unstimulated platelets. On the basis of mass concentrations, the proteases papain and tiypsin were as effective as bromelain. Using a laser thrombosis model, the in vivo effects of orally and intraveneousfy applied bromelain on thrombus formation in rat mesenteric vessels were studied. Bromelain, orally applied at 60 mg/kg body weight, inhibited the thrombus formation in a time dependent manner, the maximum being after 2 hours in 11% of arterioles and 6% of venoles. Intmveneous application at 30 mg/kg was slightly more active in reducing thrombus formation in aiterioles (13%) and venoles (5%), suggesting that oralfy applied bromelain is biologically active. These results may help to explain some of the clinical effects absented after bromelain treatment in patients with thrombosis and related diseases. Bromelain, an extract from the stem of pineapple plant, is a mixture of various basic and acidic thiol proteases and inhibitors (1, 2). As a drug, bromelain has been used for the therapy of different diseases, including thrombosis, rheumatic arthritis, inflammatory diseases such as atherosclerosis, adjuvant cancer treatment and others (for review see ref. 3). Click here to read full study.



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