Acute diuretic activity of aqueous Erica multiflora flowers and Cynodon dactylon rhizomes extracts in rats

Aim of the study

The aim of the present study is to evaluate the diuretic potential and effect on urinary electrolytes of aqueous Erica multiflora L. (Ericaceae) flowers and Cynodon dactylon L. (Poaceae) rhizomes extracts in rats.

Materials and methods

Different concentrations of these plants extract (0.125, 0.250, and 0.500 g/kg of body weight) or the reference drug furosemide (0.015 g/kg) were administrated orally to hydrated male Wistar rats and their urine output was measured at several interval of time after a single dose administration. Furthermore, a toxicological effect of both plants was undertaken as well.

Results

The results showed that furosemide induced significant diuresis and electrolytes excretion during the first hours. Plant extracts increased significantly urinary output and electrolytes excretion at the dose of 0.250 g/kg for Erica multiflora and 0.500 g/kg for Cynodon dactylon. This diuretic effect seems to be not related to K⁺ plant content. Urinary pH remained mostly unchanged during the course of the study for both plant extracts. No lethality was observed among animals when using Erica multiflora even at the dose of 10 g/kg while Cynodon dactylon, instead, caused 50% of rat death (LD50) at 4.5 g/kg.

Conclusion

We concluded that both aqueous herb extracts administered, particularly, at the dose of 0.500 g/kg induce significant effect on urinary output of water and electrolytes and justify their use as diuretic remedy in traditional medicine.     

Metabolic intervention of aflatoxin B1 toxicity by curcumin

Ethno pharmacological relevance

Curcumin, bioactive principle of turmeric (Curcuma longa Linn) is an important constituent of Indian traditional medicine. Turmeric has been known to possess several therapeutic properties.

Aim of the study

The modulatory effect of dietary curcumin (0.05%, w/w) on drug metabolizing and general marker enzymes of liver and formation of AFB₁-adducts (DNA and protein) due to dietary AFB₁ exposure for a period of 6 weeks in a rodent model, have been evaluated.

Materials and methods

Drug metabolizing enzymes CYP1A1, GSHT, UGT1A and general marker enzymes (LDH, ALT, AST, ALP and γ-GT) of liver were estimated by standardized methods. Aflatoxin adducts (DNA and protein) were quantitated by indirect competitive ELISA.

Results

Dietary curcumin enhanced GSHT (p < 0.001) and UGT1A1 (p < 0.05) activity and significantly reduced the activity of CYP1A1 (p < 0.001), in rats exposed to aflatoxin B₁. Supplementation of curcumin in the diet normalized the altered activities of LDH and ALT. At molecular level, curcumin significantly reduced AFB₁–N⁷-guanine adduct (p < 0.001) excretion in the urine, DNA adduct (p < 0.05) in the liver and albumin adduct (p < 0.001) in the serum.

Conclusion

The experimental results substantiates that curcumin intervention ameliorates the AFB₁ induced toxicity.     

Analgesic and anti-inflammatory effects of Pistacia integerrima extracts in mice

Aim of study

Extracts of Pistacia integerrima galls have been dispensed by Traditional Practitioners of Subcontinent for chest diseases as well as for aches and pains in the body. This study was planned to evaluate the possible analgesic and antiinflammatory effects of Pistacia integerrima extracts.

Materials and methods

Analgesia was determined using acetic acid induced abdominal constriction and formalin induced paw licking in mice. Antinociceptive effect was observed by thermally induced algesia in mice.

Results

Pistacia integerrima leaves extracts showed significant response against chemically induced pain (P < 0.001) whereas galls extracts had highly significant protection (P < 0.0001) in a dose dependent manner. In thermally induced algesia, Pistacia integerrima galls extracts 200 mg/kg (p.o.), showed significant (P < 0.05) response but less than pentazocine and diclofenac, positive references. The extracts of Pistacia integerrima 50–200 mg/kg (p.o.) had modest activity against hind paw acute and chronic inflammation induced by formalin (P < 0.01).

Conclusion

These results demonstrate that Pistacia integerrima extracts have antinociceptive and analgesic effects and no apparent acute toxicity on oral administration.     

Anti-angiogenic effects and mechanisms of polysaccharides from Antrodia cinnamomea with different molecular weights

Ethnopharmacological relevance

Antrodia cinnamomea is a popular medicinal mushroom in Taiwan that has been widely used for treatment of various cancers and liver diseases.

Aim of the study

This study aimed to investigate the immunomodulatory effect on angiogenesis of polysaccharides from mycelia of Antrodia cinnamomea (PMAC).

Materials and methods

PMAC were extracted in boiling water, precipitated with 95% ethanol, and separated into four different molecular weights (<5, 5–30, 30–100, >100 kDa). Tube formation and chorioallantoic membrane (CAM) assay were used to determine the in vitro and ex vivo anti-angiogenic effects.

Results

Only the PMAC-mononuclear cells (MNCs)-conditioned medium (CM) with MW > 100 kDa significantly and concentration-dependently decreased the secretion of vascular endothelial growth factor in human leukemia cells and inhibited the matrigel tube formation in human umbilical vein endothelial cells. Similarly only the PMAC-MNC-CM with MW > 100 kDa significantly and concentration-dependently increased the levels of interleukin (IL)-12 and interferon-γ (IFN-γ). In addition, the ex vivo CAM assay revealed that only the PMAC with MW > 100 kDa significantly and dose-dependently inhibited neovascularization.

Conclusions

PMAC with MW > 100 kDa are anti-angiogenic in vitro and ex vivo, and the effects are likely through immunomodulation.     

Protective effects of astragaloside IV on porcine-serum-induced hepatic fibrosis in rats and in vitro effects on hepatic stellate cells

Ethnopharmacological relevance

Astragaloside IV is the primary pure saponin isolated from Astragalus membranaceus, one of the valuable traditional medical herbs. Antifibrotic activities of Astragalus membranaceus have been extensively proved.

Aim of the study

To investigate the effects of astragaloside IV on hepatic stellate cells (HSCs) and hepatic fibrosis in rats induced by porcine-serum (PS).

Materials and methods

Liver fibrosis was induced by PS injection (0.5 ml, twice a week) for 12 weeks. Astragaloside IV (2.0, 4.0 mg kg⁻¹) was administered intragastrically. Liver samples were subjected to histological and immunohistochemical studies. In vitro effects of astragaloside IV on primary cultured HSCs were detected by incorporation assays.

Results

Astragaloside IV delayed the formation of liver fibrosis and decrease the serum levels of hyaluronic acid (HA), procollagen type III (PCIII) and hydroxyproline (Hyp) content in liver. The levels of transforming growth factor-β₁ (TGF-β₁) in serum and expression in liver were significantly decreased by astragaloside IV. Collagen synthesis and proliferation were significantly inhibited by astragaloside IV (1.5, 3.0, 6.0, 12.0 and 24.0 mg L⁻¹) in HSCs.

Conclusion

The results showed that astragaloside IV displays antifibrotic effects in rats induced by PS, the mechanism by which might be associated with its inhibitory effects on collagen synthesis and proliferation in HSCs.     

Pharmacological mechanisms underlying the vascular activities of Loranthus ferrugineus Roxb. in rat thoracic aorta

Aim of the study

The present study was aimed to investigate the pharmacological basis for the use of Loranthus ferrugineus in hypertension.

Materials and methods

Loranthus ferrugineus methanol extract (LFME) was obtained using Soxhelt extractor and then successively fractionated using chloroform, ethyl acetate and n-butanol. The n-butanol fraction of LFME (NBF-LFME) was studied using isolated rat thoracic aorta.

Results

NBF-LFME (1.0 × 10⁻⁵ to 3.0 mg/ml) was found to be the most potent to concentration-dependently relax the endothelium-intact phenyephrine (PE, 1 μM)- and high K⁺ (80 mM)-precontracted rat aortic rings. Removal of the endothelium completely abolished the vascular relaxing properties of NBF-LFME. Pretreatment with atropine (1 μM), l-NAME (10 μM), indomethacin (10 μM) and methylene blue (10 μM) significantly blocked NBF-LFME-mediated relaxation. Endothelium-dependent and -independent relaxations induced by acetylcholine (ACh) and sodium nitroprusside (SNP), respectively, were significantly enhanced in aortic rings pretreated with NBF-LFME when compared to those observed in control aortic rings. On the contrary, glibenclamide (10 μM), propranolol (1 μM) and prazosin (0.01 μM) did not alter NBF-LFME-induced relaxation.

Conclusions

The results suggest that NBF-LFME induced vascular relaxation by stimulating muscarinic receptors, activating the endothelium-derived nitric oxide-cGMP-relaxant pathway, promoting prostacyclin release and/or possibly through its ability to lengthen the released nitric oxide half-life. The present data further supports previous in vivo findings and explain the traditional use of Loranthus ferrugineus as an anti-hypertensive agent.     

Fresh ginger (Zingiber officinale) has anti-viral activity against human respiratory syncytial virus in human respiratory tract cell lines

Ethnopharmacological relevance

Ginger, Zingiber officinale Roscoe, is a common spice and also a widely used medicinal plant in ancient China. Ginger is an ingredient of Ge-Gen-Tang (Kakkon-to; GGT). GGT has been proved to have antiviral activity against human respiratory syncytial virus (HRSV). However, it is unknown whether ginger is effective against HRSV.

Aim of the study

To find a readily available agent to manage HRSV infection, the authors tested the hypothesis that ginger can effectively decrease HRSV-induced plaque formation in respiratory mucosal cell lines.

Materials and methods

Effect of hot water extracts of fresh and dried gingers on HRSV was tested by plaque reduction assay in both human upper (HEp-2) and low (A549) respiratory tract cell lines. Ability of ginger to stimulate anti-viral cytokines was evaluated by enzyme-linked immunosorbent assay (ELISA).

Results

Fresh ginger dose-dependently inhibited HRSV-induced plaque formation in both HEp-2 and A549 cell lines (p<0.0001). In contrast, dried ginger didn't show any dose-dependent inhibition. 300 μg/ml fresh ginger could decrease the plaque counts to 19.7% (A549) and 27.0% (HEp-2) of that of the control group. Fresh ginger was more effective when given before viral inoculation (p<0.0001), particularly on A549 cells. 300 μg/ml fresh ginger could decrease the plaque formation to 12.9% when given before viral inoculation. Fresh ginger dose-dependently inhibited viral attachment (p<0.0001) and internalization (p<0.0001). Fresh ginger of high concentration could stimulate mucosal cells to secrete IFN-β that possibly contributed to counteracting viral infection.

Conclusions

Fresh, but not dried, ginger is effective against HRSV-induced plaque formation on airway epithelium by blocking viral attachment and internalization.     

Antibacterial activity of the roots,stems and leaves of Alchornea floribunda

Ethnopharmacological relevance

Alchornea floribunda Müll. Arg. is used in traditional medicine across Africa for the treatment of bacterial, fungal, parasitic and inflammatory disorders.

Aim of the study

To evaluate the antibacterial activity of the crude extracts of different plant parts in order to provide a scientific rationale for the proposed broad efficacy of Alchornea floribunda in the treatment of bacterial infections.

Materials and methods

Extracts of roots, stems and leaves were prepared using solvents of various polarities in order to extract a wide range of phytochemicals. The antibacterial activity of these crude extracts was evaluated by micro-dilution assay, against Gram-positive (i.e. Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus saprophyticus) as well as Gram-negative (i.e. Escherichia coli, Klebsiella pneumoniae, Moraxella catarrhalis and Proteus mirabilis) bacteria.

Results

Generally, the ethanol (EtOH), methanol (MeOH), ethyl acetate (EtOAc) and chloroform (CHCl₃) extracts demonstrated the best activities, with the leaves exhibiting the highest average activity for six of the eight pathogens. Of these, the ethanolic leaf extract was the most active against Staphylococcus aureus with an MIC value of 50 µg/mL. Some other notable activity was observed for the ethyl acetate and chloroform root extracts against Staphylococcus aureus (50 µg/mL), and for selected stem extracts against Staphylococcus aureus (50 µg/mL), Klebsiella pneumoniae (63 µg/mL) and Staphylococcus saprophyticus (63 µg/mL).

Conclusion

This study demonstrates the promising antibacterial activity of Alchornea floribunda against both Gram-positive and Gram-negative bacteria responsible for gastrointestinal, skin, respiratory and urinary ailments, and validates its use in the ethnopharmacology of the region.     

Macrophage immunomodulatory activity of the polysaccharides from the roots of Bupleurum smithii var. parvifolium

Aim of the study

Radix Bupleuri, one of the most frequently prescribed crude herbs in traditional Chinese medicine, has been used for centuries to treat inflammatory diseases. However, little is known about the therapeutic mechanisms of crude polysaccharides (BPs) isolated from the roots of Bupleurum smithii var. parvifolium. Macrophages play important roles in inflammatory diseases such as systemic lupus erythematosus (SLE). The purpose of the present work was to investigate immunomodulative effects of Bupleurum polysaccharides on murine peritoneal macrophages.

Materials and methods

BALB/c mice were administered intragastrically with Bupleurum polysaccharides 20, 40, and 80 mg kg⁻¹ day⁻¹, or prednisone 3 mg kg⁻¹ day⁻¹ or levamisole 25 mg kg⁻¹ day⁻¹ from day 0 to day 6. Peritoneal macrophages were isolated 5 days after intraperitoneal injection of 1 mL 5% sodium thioglycollate. Phagocytic functions of macrophages were studied; cytokines concentrations in the culture supernatants were determined by enzyme-linked immunosorbent assay and the secretion of nitric oxide (NO) was quantified by Griess reaction.

Results

Treatment with BPs enhanced phagocytic functions of macrophages (phagocytosis of apoptotic thymocytes, IgG-opsonized sheep red blood cells and chicken red blood cells) and inhibited LPS-induced productions of NO and proinflammatory cytokines (interleukin-1β, interleukin-6 and tumor necrosis factor-α).

Conclusions

Bupleurum polysaccharides up-regulated phagocytic activities but inhibited LPS-induced productions of proinflammatory mediators. These data suggested that at least part of the traditional beneficial effects of Bupleurum on inflammatory diseases could be ascribed to the immunomodulatory effects of Bupleurum polysaccharides on macrophages.     

Hypoglycaemic effects of Mammea africana (Guttiferae) in diabetic rats

Aim of the study

The stem bark of Mammea africana Sabine (Guttiferae) is used in African rain forest to treat various diseases, including diabetes mellitus. We investigated whether Mammea africana extract induced hypoglycaemic activity in rats.

Materials and methods

We tested the effects of acute (5 h) and sub-acute (21 days) oral administrations of the CH₂Cl₂–MeOH stem bark extract of Mammea africana (19–300 mg/kg body weight) on blood glucose levels of normal and streptozotocin (STZ)-induced type 1 diabetic rats. The effects were compared with those of glibenclamide.

Results

Acute administration reduced blood glucose in the diabetic rats only (33.87%, P < 0.01). Sub-acute treatment for 21 days also reduced blood glucose level in diabetic rats (73.29%, P < 0.01). A reduction or stabilization in total serum protein, triglyceride, cholesterol and alanine amino transferase levels was also observed. No effect was observed on body weight loss but food and water intakes were significantly reduced (P < 0.01) in diabetic rats. The maximal anti-diabetic effect was obtained with the dose of 75 mg/kg and was more important than that of glibenclamide.

Conclusion

It can be concluded that extracts of Mammea africana exhibited a significant anti-hyperglycaemic activity and improved the metabolic alterations in STZ-diabetic rats. These results provide a rationale for the use of Mammea africana to treat diabetes mellitus and hypercholesterolemia.     

Mouririelliptica: Validation of gastroprotective,healing and anti-Helicobacter pylori effects

Ethnopharmacological relevance

Mouriri elliptica Martius (Melastomataceae) is species reputed in folk medicine to heal gastric ulcer and gastritis.

Aim of the study

Methanolic extract (ME) and ethyl acetate fraction (EAF) from leaves of Mouriri elliptica were evaluated for their gastroprotective, healing, immunological, toxicological and anti-Helicobacter pylori activities.

Material and methods

The gastroprotective action of ME and EAF was evaluated in rodent experimental models and to elucidate mechanisms of action, the antisecretory action, involvements of NO, SH, PGE₂, anti-Helicobacter pylori action of ME was evaluated. We also used immunohistochemical (PCNA and COX-2) and immunomodulatory (murine peritoneal macrophages) assays to evaluate Mouriri elliptica effects.

Results

ME present gastroprotective action without antisecretory effect. Otherwise, ME showed anti-Helicobacter pylori action (MIC = 0.025 μg/mL) and was able to inhibit NO production by macrophages. This species also accelerate the healing of ulcerated gastric mucosa by stimulating proliferation factors (PCNA), COX-2 and maintained basal PGE₂ level independent action of NSAID in gastric mucosa. The phytochemical investigation showed that this species possesses phenolic acid derivatives, acylglycoflavonoids and condensed tannins which probably influenced their pharmacological action.

Conclusion

All these results suggest the efficacy and safety of Mouriri elliptica in combating and healing gastric ulcer.     

Anti-inflammatory intestinal activity of Abarema cochliacarpos (Gomes) Barneby & Grimes in TNBS colitis model

Aim of the study

To assess the anti-inflammatory effect of butanolic fraction of methanolic extract from bark of Abarema cochliacarpos in acute ulcerative colitis model induced by intracolonic administration of trinitrobenzene sulfonic acid (TNBS) in Wistar rats.

Materials and methods

Abarema cochliacarpos (100 and 150 mg/kg/day) was administered by gavage 48, 24 and 1 h prior to the induction of colitis with 10 mg/kg of TNBS and, 24 h later.

Results

Phytochemical studies by mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) revealed that catechins were a major component into condensate class of tannins. Treatment with Abarema cochliacarpos decreased significantly macroscopic damage as compared with TNBS (p < 0.05). Histological analysis showed that both doses of the extract improved the microscopic structure and preserved some areas of the colonic mucosa structure. In addition, myeloperoxidase activity (MPO), as a marker of neutrophil infiltration, was decreased in a dose-dependent way (p < 0.01 and p < 0.001 respectively), TNF-α level was also diminished with the highest dose of the extract (p < 0.001) and, IL-10 level obtained no significant results. In order to elucidate some of the mechanisms, expression of inducible inflammatory enzymes, such as cyclooxygenase (COX)-2 and nitric oxide synthase (iNOS), were studied showing a significant reduction. Finally, the involvement of c-Jun N-terminal kinase (JNK) signalling demonstrated a reduction in the JNK activation with the highest dose (p < 0.05 vs TNBS).

Conclusions

We have shown for the first time that the extracts obtained from Abarema cochliacarpos bark possess active substances, which exert marked protective effects in acute experimental colitis, confirming and justifying, at least in part, the popular use of this plant to treat gastrointestinal diseases.     

Anti-influenza virus effects of the aqueous extract from Mosla scabra

Ethnopharmacological relevance

Mosla scabra (Thunb.) C. Y. Wu is a broadly used species in the southeastern China as an antipyretic and antiviral drug for the treatment of colds, fever, inflammation and chronic bronchitis.

Aim of the study

To investigate the anti-influenza virus activities of the aqueous extract from Mosla scabra (AEMS), and to provide evidence for the implication of its immunomodulatory potential for the overall protective effect in lethal murine experimental influenza A infection.

Materials and methods

The anti-influenza virus activities of AEMS or ribavirin were evaluated in embryonated eggs and in a mouse infection model and the effects of AEMS on early immune responses during influenza virus infection were evaluated in a mouse infection model.

Results

At the concentration of 0.3–30.0 mg/ml, AEMS exhibited both preventive and therapeutical effect on embryonated egg. Oral administration of AEMS to mice infected with influenza virus A (IVA) was highly effective in preventing death, inhibiting pneumonia and reducing lung viral titers. It also significantly enhanced IFN-γ, IL-2, IL-4 and IL-5 cytokine-producing splenocytes, increased T-cell subsets like CD4⁺ and CD4⁺/CD8⁺, decreased levels of IL-6 after infection, and provided protective immunity. At a dosage of 4800 mg/kg, almost all treated mice survived, suggesting that AEMS is of low toxicity. However, ribavirin has weaker efficacy compared to AEMS.

Conclusions

These results showed, for the first time, while both AEMS and ribavirin appeared to have similar efficacy against IVA, AEMS playing a role as an immunomodulator and antiviral inhibitor during influenza virus infection, was considered to be less toxic and may warrant further evaluation as a possible agent for the treatment of influenza.     

Effects on the expression of GABAA receptor subunits by jujuboside A treatment in rat hippocampal neurons

Aim of study

To determine the effect of jujuboside A (JuA) in modulating the γ-aminobutyric acid (GABA_A) receptor subunits gene expression of hippocampal neurons at different terms in vitro.

Materials and methods

Hippocampal neurons of rat were cultured in vitro, treated with JuA or diazepam (DZP). Then GABA_A receptor mRNAs were evaluated by semi-quantitative RT-PCR.

Results

JuA at the low dose of 41 μM (about 0.05 g/l) induced significant increase of GABA_A receptor α1, α5, β2 subunit mRNAs in both 24 and 72 h treatments. JuA at the high dose of 82 μM (about 0.1 g/l) significantly increased GABA_A receptor α1, α5 subunit mRNA levels and decreased β2 subunit mRNA level at 24 h treatment, and decreased GABA_A receptor subunit α1, β2 mRNAs expression at 72 h treatment. DZP of 10 μM significantly increased expression of GABA_A receptor subunit α1, α5 and decreased expression of β2 at 24 h treatment, and decreased α1, α5, β2 subunits gene expression at 72 h treatment.

Conclusion

Differences in alterations in GABA_A receptor subunit mRNAs expression following JuA and DZP treatments could help to explain the differences in the pharmacological action of the two drugs.     

Anti-inflammatory and antinociceptive effects of hydroalcoholic extract from Pseudobombax marginatum inner bark from caatinga potiguar

Ethnopharmacological relevance

The Pseudobombax marginatum (St Hil) Rob., Malvaceae, is mentioned in ethnobotanical studies. It is used as anti-inflammatory, for ulcers and gastritis, and back pain. To evaluate anti-inflammatory and antinociceptive activities a hydroalcoholic extract (HE) from inner bark was prepared.

Materials and methods

For the anti-inflammatory activity, carrageenan-induced paw edema and peritonitis models, and also myeloperoxidase assay were used. For the antinociceptiva activity acetic acid-induced writhing, hot plate and formalin tests were employed.

Results

The HE extract exhibited an intense inhibition in carrageenan-induced edema model and also in myeloperoxidase activity at the doses of 100 and 300 mg/kg. The leukocyte migration into the peritoneal cavity was also inhibited at the doses of 30, 100 and 300 mg/kg. A similar profile was observed against acid-induced abdominal contortions and in formalin second phase test at the doses of 30 and 100 mg/kg, but this treatment did not affect the behavior of animals in the hot plate test.

Conclusions

The experimental data of the HE from Pseudobombax marginatum show anti-inflammatory and antinociceptiva activities, confirming the indication from traditional medicine; however further studies are required to define and isolate the active anti-inflammatory and antinociceptiva components from this active specie.     

Studies on the analgesic,anti-inflammatory and antipyretic effects of Parquetina nigrescens leaf extract

Ethnopharmacological relevance

Parquetina nigrescens is a shrub that is commonly used in different parts of West Africa for the treatment of several ailments which includes pain, fever and inflammatory conditions.

Aim of the study

The present study was designed to investigate the analgesic, anti-inflammatory and antipyretic effects of the aqueous extract of Parquetina nigrescens leaves in rats.

Materials and methods

Five groups were used for each study, groups 1 and 5 served as control (saline) and reference (indomethacine) respectively, while groups 2–4 received the extract (50–200 mg/kg) orally. Formalin paw licking and hot plate latency tests were used for analgesic studies. Carrageenan oedema, cotton pellet granuloma and formaldehyde arthritis models were used to quantify the anti-inflammatory activities while the brewer’s yeast was used for inducing pyrexia.

Results

The results of the analgesic study show that the extract produced significant (p < 0.05) analgesia in the hot plate and in the formalin tests. In the anti-inflammatory study, Parquetina nigrescens produced significant (p < 0.05) inhibition of the various types of inflammation. The extract also inhibited the pyrexia induced by brewer’s yeast.

Conclusion

The result justifies the traditional uses of Parquetina nigrescens for the treatment of fever, inflammatory and painful conditions.     

An in vitro study on antiproliferative and antiestrogenic effects of Boerhaavia diffusa L. extracts

Ethnopharmacological relevance

Boerhaavia diffusa L. (Nyctinaceae) is a plant of tropical region used in Indian traditional medicine for the treatment of human ailments including abdominal tumor, jaundice, dyspepsia, menstrual disorders, etc. This plant also has antilymphoproliferative, antimetastatic and immunomodulatory effects.

Aim of the study

This study aimed to assess the antiproliferative and antiestrogenic properties of methanol extract of Boerhaavia diffusa (BME) in MCF-7 breast cancer cell lines.

Materials and methods

The effective concentration range of BME on cell viability was analyzed using MTT assay. Hydroxylapatite assay (HAP) was carried out to confirm the competitive binding of BME to the estrogen receptor (ER). The effect of BME on the expression of a selected estrogen responsive gene pS2 was analyzed by RT-PCR. The ability of BME to alter the cell cycle phases and distributions were studied using FACS analysis.

Results

Treatment with varying concentrations of BME (20–320 μg/mL) resulted in moderate to very strong growth inhibition in MCF-7 cell lines. BME competed with [³H]-estradiol for binding to ER with IC₅₀ value of 320 ± 25 μg/mL. RT-PCR analysis revealed that BME reduced the mRNA expression of pS2 indicating the antiestrogenic action of BME. BME treatment for 48 h resulted in a remarkable increase in the number of MCF-7 cells in the G0-G1 fraction from 69.1% to 75.8%, with a reciprocal decrease of cells in all other phases indicating cell cycle arrest at G0-G1 phase.

Conclusions

The results demonstrate that Boerhaavia diffusa possess antiproliferative and antiestrogenic properties and suggest that it may have therapeutic potential in estrogen dependent breast cancers.     

Identification of the protein components displaying immunomodulatory activity in aged garlic extract

Ethnopharmacological relevance

Traditionally, garlic (Allium sativum L.; Alliaceae) has been known to boost the immune system. Aged garlic has more potent immunomodulatory effects than raw garlic. These effects have been attributed to the transformed organosulfur compounds; the identity of the immunomodulatory proteins in aged garlic extract (AGE) is not known.

Aim of the study

The major aims are to examine the changes occurring in the protein fraction during ageing of garlic and to identify the immunomodulatory proteins.

Materials and methods

Changes occurring in garlic during ageing have been examined by protein quantitation and gel electrophoresis. Purification and identification of the immunomodulatory proteins have been achieved by Q-Sepharose chromatography and mitogenic activity.

Results

Only two major proteins (12–14 kDa range by SDS-PAGE) are observed in AGE. The purified protein components QA-1, QA-2, and QA-3 display immunomodulatory and mannose-binding activity; QA-2 shows the highest mitogenic activity. The identity of QA-2 and QA-1 proteins with the garlic lectins ASA I and ASA II, respectively, has been confirmed by hemagglutination analysis. QA-3 exhibits mitogenic activity, but no hemagglutination activity.

Conclusions

The immunomodulatory activity of AGE is also contributed by immunomodulatory proteins. The major immunomodulatory proteins have been identified as the well-known garlic lectins.     

Immunomodulatory effects of crude phenylethanoid glycosides from Ligustrum purpurascens

Ethnopharmacological relevance

Ligustrum purpurascens, named as “Ku ding cha”, has been used as a kind of functional tea in southern China for about two thousand years, which has the effects on diuresis, anti-hypertension, weight-loss and anti-inflammation.

The aim of the study

This study was aimed to investigate the immune enhancement effects of the crude phenylethanoid glycosides (CPGs) from Ligustrum. Purpurascens on mice and analyze the chemical profiles of phenylethanoid glycosides in the CPGs.

Materials and methods

The immune functions enhancing potential of CPGs was determined using serum hemolysin antibody, phagocytosis, splenocyte antibody production, and NK cells activity assays. The contents of five major constituents in the crude glycosides of Ligustrum purpurascens were determined by using liquid chromatography, other five glycosides were deduced according to their UV and MS spectra compared with the literature as well.

Results

In the immunizing experiment, mice treated with different doses of CPGs showed an increase (p<0.01) in the haemagglutination titre compared with the control group. The increases (p<0.05) were found to be significant at doses of 440 mg/kg and 1.32 g/kg in the experiments of antibody production of spleen cells, MΦ phagocytosis of chicken RBCs and NK cell activity. Further chemical characterization yielded 10 constituents from CPGs, five glycosides were quantified by HPLC and the structures of other five compounds were speculated according to their UV and MS spectra.

Conclusion

The results suggested that phenylethanoid glycosides from Ligustrum purpurascens have immunomodulatory effects on mice.     

Evidence of the mechanism of action of Erythrina velutina Willd (Fabaceae) leaves aqueous extract

Ethnopharmacological relevance

Erythrina velutina is traditionally used for sleepiness, convulsions and nervous system excitation in Brazil. Although central effects have been reported for Erythrina velutina, little is known about its mechanism of action.

Aim of the study

To investigate the pharmacological evidences of mechanism of action of Erythrina velutina leaves aqueous extract (AE).

Materials and methods

Terminal segments of the guinea-pig ileum (n = 5–8) were mounted in an organ bath and isotonic contractions were recorded. Phytochemical screening was carried out on AE.

Results

AE (0.025–2.50 mg/ml) produced contractile response in the guinea-pig ileum, yielding typical concentration–response curves (EC₅₀ = 0.63 mg/ml). Electrically evoked contractions were significantly increased in the presence of AE. AE-elicited contractions were significantly reduced by bicuculline, tetrodotoxin, atropine, verapamil or incubation in low calcium–high potassium solution. Atropine along with verapamil abolished AE contractile response. Alkaloids, catechins, steroids, flavonols, flavononols, flavonoids, phenols, saponins, tannins, triterpenoids, and xanthones were detected in AE.

Conclusions

AE contains important constituents for pharmacological activities. AE-induced contractions seem to involve GABA_A receptor activation, acetylcholine release, muscarinic receptor activation, augmentation of Ca²⁺ entry through L-type calcium channels, and calcium release from the intracellular stores. These findings provide further support for Erythrina velutina traditional uses.