In vitro anti-breast cancer activity of ethanolic extract of Wrightia tomentosa: Role of pro-apoptotic effects of oleanolic acid and urosolic acid

Ethnopharmacological relevance

Wrightia tomentosa Roem. & Schult. (Apocynaceae) is known in the traditional medicine for anti-cancer activity along with other broad indications like snake and scorpion bites, renal complications, menstrual disorders etc. However, the anti-cancer activity of this plant or its constituents has never been studied systematically in any cancer types so far.

Aim of the study

To evaluate the anti-cancer activities of the ethanolic extract of W. tomentosa and identified constituent active molecule(s) against breast cancer.

Material and methods

Powdered leaves of W. tomentosa were extracted with ethanol. The ethanolic extract, subsequent hexane fractions and fraction F-4 of W. tomentosa were tested for its anti-proliferative and pro-apoptotic effects in breast cancer cells MCF-7 and MDA-MB-231.

Results

The ethanolic extract, subsequent hexane fractions and fraction F-4 of W. tomentosa inhibited the proliferation of human breast cancer cell lines, MCF-7 and MDA-MB-231. The fraction F-4 obtained from hexane fraction inhibited proliferation of MCF-7 and MDA-MB-231 cells in concentration and time dependent manner with IC₅₀ of 50 μg/ml and 30 μg/ml for 24 h, 28 μg/ml and 22 μg/ml for 48 h and 25 μg/ml and 20 μg/ml for 72 h respectively. The fraction F-4 induced G1 cell cycle arrest, reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential and subsequent apoptosis. Apoptosis is indicated in terms of increased Bax/Bcl-2 ratio, enhanced Annexin-V positivity, caspase 8 activation and DNA fragmentation. The active molecule isolated from fraction F-4, oleanolic acid and urosolic acid inhibited cell proliferation of MCF-7 and MDA-MB-231 cells at IC₅₀ value of 7.5 μM and 7.0 μM respectively, whereas there is devoid of significant cell inhibiting activity in non-cancer originated cells, HEK-293. In both MCF-7 and MDA-MB-231, oleanolic acid and urosolic acid induced cell cycle arrest and apoptosis as indicated by significant increase in Annexin-V positive apoptotic cell counts.

Conclusion

Our results suggest that W. tomentosa extracts has significant anti-cancer activity against breast cancer cells due to induction of apoptosis pathway. Olenolic and urosolic acid are important constituent molecules in the extract responsible for anti-cancer activity of W. tomentosa.     

In vitro and in vivo antioxidant effects of the ethanolic extract of Swertia chirayita

Ethnopharmacological relevance

Swertia chirayita, a medicinal herb endemic to the Tibetan region, is used as a special remedy for liver disorders. The hepatoprotective activity of its plant extracts has been associated with its antioxidant activity. This paper aims to investigate the in vitro and in vivo antioxidant effects of Swertia chirayita extracts (SCE).

Materials and methods

Antioxidant ability of Swertia chirayita was investigated by employing several established in vitro methods. In vivo antioxidant activity was tested against CCl₄-induced toxicity in mice. The levels and activities of malondialdehyde (MDA) and antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), were then assayed using standard procedures.

Results

SCE exhibited strong antioxidant ability in vitro. The liver and kidney of CCl₄-intoxicated animals exhibited a significant (p < 0.001) decrease in SOD, CAT, and GSH levels. Additionally, these organs exhibited a significant (p < 0.001) increase in MDA level. CCl₄ did not exhibit toxicity on mice treated with SCE and Vitamin E. The effects of Swertia chirayita (three dosages) were comparable to those of Vitamin E, except in MDA level in the liver and GSH level in the kidney (p < 0.05).

Conclusion

This study suggests that the ethanolic extract of Swertia chirayita possesses in vitro and in vivo antioxidant effects. This supports the traditional use of Swertia chirayita in Tibetan medicine to cure liver diseases.     

In vitro and in vivo antioxidant activity of the ethanolic extract from Meconopsis quintuplinervia

Aims of the study

Meconopsis quintuplinervia, a medicinal herb endemic to the Tibetan region, is used to treat hepatitis. The aim of this study is to evaluate the antioxidant potential of the ethanolic extract of this herb using different assays.

Materials and methods

The antioxidant capacity of Meconopsis quintuplinervia was investigated using various established in vitro systems. An in vivo study of carbon tetrachloride (CCl₄)-induced antioxidant activity in mice was also conducted by examining the levels of malondialdehyde (MDA) and the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH).

Results

The extract showed strong in vitro antioxidant ability. In the in vivo study, CCl₄-induced oxidative stress caused significant decreases in the SOD, CAT, and GSH levels and a significant increase in the MDA level, most of which were significantly reversed (except for SOD in the liver.) by treatment with the extract and standard Vitamin E.

Conclusion

This study clearly indicates that the ethanolic extract of Meconopsis quintuplinervia is a valuable source of natural antioxidants. These findings provide scientific support for the traditional use of this herb as a Tibetan medicine for liver diseases.     

Antimalaria activity of ethanolic extract of Tetrapleura tetraptera fruit

The in vivo antiplasmodial activity of the ethanol fruit extract of Tetrapleura tetraptera used as spice and in the treatment of various ailment in Niger Delta region of Nigeria was evaluated in Plasmodium berghei infected mice. Tetrapleura tetraptera (300-900 mg/kg day) exhibited significant (P < 0.05) blood schizonticidal activity both in 4-day early infection test and in established infection with a considerable mean survival time comparable to that of the standard drug, chloroquine, 5 mg/kg day. The fruit extract possesses significant (P < 0.05) antiplasmodial activity with may have contributed to the immune status of the Nigerians against malaria in addition to its nutritive value.     

Antidiabetic effect of Ficus religiosa extract in streptozotocin-induced diabetic rats

Aims of study

In Indian traditional system of medicine, Ficus religiosa (Family Moraceae) is prescribed for the treatment of diabetes mellitus. In the present study, the antidiabetic effect of aqueous extract of Ficus religiosa bark (FRAE) was investigated in normal, glucose-loaded hyperglycemic and streptozotocin (STZ)-induced diabetic rats.

Materials and methods

Oral administration of FRAE at the doses of 25, 50 and 100 mg/kg was studied in normal, glucose-loaded and STZ-diabetic rats.

Results

The three doses caused significant reduction in blood glucose levels in all the models. The effect was more pronounced in 50 and 100 mg/kg than 25 mg/kg. FRAE also showed significant increase in serum insulin, body weight and glycogen content in liver and skeletal muscle of STZ-induced diabetic rats while there was significant reduction in the levels of serum triglyceride and total cholesterol. FRAE also showed significant antilipidperoxidative effect in the pancreas of STZ-induced diabetic rats. The antidiabetic effect of Ficus religiosa was compared with glibenclamide, a well-known hypoglycemic drug.

Conclusion

The results indicate that aqueous extract of Ficus religiosa bark possesses significant antidiabetic activity.     

Anti-diabetic activity of Chiliadenus iphionoides

Ethnopharmacological relevance

Chiliadenus iphionoides (Boiss. & Blanche) Brullo (Asteraceae), a small aromatic shrub found throughout Israel, is used traditionally in the treatment of diabetes mellitus. In this study, Chiliadenus iphionoides anti-diabetic activity was characterized using cellular and animal models.

Materials and methods

Pancreatic β cells, adipocytes, and skeletal myotubes were treated with an ethanolic extract of Chiliadenus iphionoides to study the extract's effects on insulin secretion and glucose uptake. The sand rat (Psammomys obesus) was used to study Chiliadenus iphionoides acute and long term effects in vivo. An oral starch tolerance test was performed as well as a 30 day feeding study.

Results

Chiliadenus iphionoides extract increased insulin secretion in β cells as well as glucose uptake in adipocytes and skeletal myotubes. The extract also displayed hypoglycemic activity in the diabetic sand rat.

Conclusions

Chiliadenus iphionoides exhibits considerable anti-diabetic activity, although the mechanism of action remains to be determined.     

Antidiabetic effects of Justicia spicigera Schltdl (Acanthaceae)

Ethnopharmacological importance

Justicia spicigera is a plant species used for the Teenak (Huesteca Potosina) and Mayan (Yucatan peninsula) indigenous for the empirical treatment of diabetes, infections and as stimulant.

Aim of the study

To evaluate the cytotoxicity, antioxidant and antidiabetic properties of J. spicigera.

Materials and methods

The effects of ethanolic extracts of J. spicigera (JSE) on the glucose uptake in insulin-sensitive and insulin-resistant murine 3T3-F442A and human subcutaneous adipocytes was evaluated. The antioxidant activities of the extract of JSE was determined by ABTS and DPPH methods. Additionally, it was evaluated the antidiabetic properties of JSE on T2DM model.

Results

JSE stimulated 2-NBDG uptake by insulin-sensitive and insulin-resistant human and murine adipocytes in a concentration-dependent manner with higher potency than rosiglitazone 1 mM. JSE showed antioxidant effects in vitro and induced glucose lowering effects in normoglycemic and STZ-induced diabetic rats.

Conclusion

The antidiabetic effects of administration of J. spicigera are related to the stimulation of glucose uptake in both insulin-sensitive and insulin-resistant murine and human adipocytes and this evidence justify its empirical use in Traditional Medicine. In addition, J. spicigera exerts glucose lowering effects in normoglycemic and STZ-induced diabetic rats.     

Antinociceptive activity of ethanolic extract and isolated compounds of Urtica circularis

Ethnopharmacological relevance

Urtica circularis (Hicken) Sorarú is a medicinal plant commonly used in traditional medicine to relieve pain in inflammatory processes.

Aim of the study

In the present study, the in vivo antinociceptive effect of Urtica circularis ethanolic extract and its isolated compounds has been investigated.

Materials and methods

Antinociceptive activity was evaluated through writhing, formalin and hot plate tests in mice. The phytochemical analysis was performed.

Results

The extract produced significant inhibition on nociception induced by acetic acid (ED50: 72.2 mg/kg, i.p.) and formalin (ED50: 15.8 mg/kg, i.p.) administered intraperitoneally and also orally. Atropine diminished the activity of the extract in the acetic acid test. In this model, at dose of 10 mg/kg i.p., vitexin was the most active of the isolated compounds (inhibition of 91%), and chlorogenic acid, caffeic acid and vicenin-2 (6,8-di-C-glucosyl apigenin) produced an inhibition of 72%, 41% and 41%, respectively, whereas apigenin did not show any activity.

Conclusions

These results suggest that Urtica circularis extract produced antinociception possibly related to the presence of vitexin, chlorogenic, caffeic acid and vicenin-2. The activation of cholinergic systems seems to be involved in the mechanism of antinociception of the extract.     

In vitro and in vivo immunomodulating and immunorestorative effects of Astragalus membranaceus

Astragalus membranaceus is a common traditional Chinese medicinal plant widely used as a tonic to enhance the body's natural defense mechanisms. In this study, bioactive fractions were isolated from the roots of Astragalus membranaceus. One of these fractions, designated as AI, was found to be the most potent with respect to its mitogenicity on murine splenocytes. Effects of AI on both specific and nonspecific immunity in mouse models were examined. Results showed that AI could exhibit mitogenic and co-mitogenic activities on mouse splenocytes, both in vitro and in vivo. Experiments in human cell culture demonstrated that AI was also active on human lymphocytes. It was found that AI was mitogenic to T cell depleted population but virtually inactive on B cell depleted population. Intraperitoneal injection of AI into mice markedly augmented the antibody response to sheep red blood cells. Besides, both the influx of macrophages into the peritoneal cavity and the phagocytic activity of macrophages were found to be enhanced by AI in vivo. On the other hand, AI could significantly increase the interleukin-2 receptor expression on mouse splenocytes in vitro. In terms of immunorestorative activity, it was found that AI could restore the lymphocyte blastogenic response of the older mice to values that are normally found in the younger mice. Moreover, administration of AI in vivo could partially restore the depressed immune functions in tumour-bearing mice and cyclophosphamide-treated mice. Collectively, the results clearly showed that AI could exhibit immunomodulating and immunorestorative effects, both in vitro and in vivo.     

In vitro and in vivo toxicological evaluation of extract and fractions from Baccharis trimera with anti-inflammatory activity

Ethnopharmacological relevance

Baccharis trimera (Less) DC. (Asteraceae), popularly known in Brazil as “carqueja”, have been used in folk medicine to treat gastrointestinal, hepatic and renal diseases, and inflammatory processes as rheumatism.

Aim of the study

To evaluate the in vitro and in vivo toxicological effects of anti-inflammatory Baccharis trimera aqueous extract and fractions.

Materials and methods

Aqueous extract of Baccharis trimera (AEBt) was produced by infusion in boiling water. After lyophylization AEBt was extracted with 80% ethanol, originating the ethanolic supernatant fraction (EFBt) and the aqueous sediment fraction (AFBt). Anti-inflammatory properties of AEBt, EFBt or AFBt (3, 30 or 300 μg/kg b.w.) were evaluated by the carrageenan-induced mouse paw edema using indomethacin (10 mg/kg) as positive control. The growth of rat hepatoma cells (HTC) and human embryo kidney epithelial cells (HEK) was determined by protein staining assay. Cytotoxicity was assayed by the tetrazolium salt (MTT) reduction. Cyclosporin was used as reference cytotoxic drug for spleen cells and doxorubicin for HTC and HEK cells. For in vivo toxicological evaluation SW male mice were daily and oral (gavage) treated with extract/fractions at 4.2 mg/kg or 42 mg/kg during 15 days. After treatment liver or kidney cells were submitted to comet assay to determine the DNA damage index, and the glutathione S-transferase activity was assayed towards ETHA (class Pi) and CDNB (several classes). Mutagenicity was evaluated by the Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102.

Results

The anti-inflammatory effects of EFBt were higher than those of AEBt or AFBt. Mice treatment (3-300 μg/kg) with AFBt reduced the paw edema (3 h) at lower levels (29.2-37.3%; P < 0.01), than those observed for AEBt (44.7-54.2%; P < 0.001), EFBt (49.3-58.2%; P < 0.001) or indomethacin (64.6%, P < 0.001, 10 mg/kg). The growth of kidney cells (HEK) was inhibited by AEBt (IC₅₀ 182.6 μg/ml), EFBt (IC₅₀ 78.1 μg/ml) and AFBt (IC₅₀ 86.2 μg/ml), with lower effects on HTC hepatic cell (IC₅₀ 308.8 μg/ml, 396.5 μg/ml and 167.9 μg/ml, respectively). As evaluated by MTT test, AFBt exhibited cytotoxicity for HEK cells (IC₅₀ 372.5 μg/ml), but none for HTC ones; by the way, AFBt stimulated spleen cells (EC₅₀ 2.2 μg/ml) while cyclosporine, a cytotoxic reference drug inhibited them with IC₅₀ of 0.42 μg/ml; the IC₅₀ for doxorubicin for HEK and HTC cells was 0.28 μg/ml and 14.4 μg/ml, respectively, at 96 h. No mutagenic potential was observed. Mice treatment with AEBt or AFBt at 42 mg/kg for 15 days altered the kidney relative weight, but not at 4.2 mg/kg. Baccharis trimera did not change liver, spleen or popliteal lymph node relative weight. DNA damage index of kidney cells was observed on mice treated with AEBt/AFBt, but not on animals treated with EFBt, while DNA lesions were detected on liver cells only after AFBt treatment. The general activities of hepatic GST and Pi GST were reduced by EFBt and AFBt treatment, respectively.

Conclusions

Baccharis trimera did not show mutagenicity, inhibited the GST activity, a hepatic detoxification enzyme, and induced in vivo (genotoxicity) and in vitro toxicological effects to kidney cells.     

Anti-inflammatory effects of ethanolic extract and alkamides-derived from Heliopsis longipes roots

Ethnopharmacological relevance

Heliopsis longipes (A. Gray) Blake (Asteraceae) is a broadly used species in the Mexican, Central and South American Traditional Medicine for its anaesthetic, analgesic, anti-inflammatory and anti-ulcerative properties. The ethanolic extract contains alkamides, mainly affinin (spilanthol). This family of compounds exerts an in vitro inhibitory action on the cyclooxygenase and lipoxygenase enzymes.

Aim of the study

The present study approaches the anti-inflammatory effect of the extract and its main bioactive component affinin and derived isobutyl-decanamide.

Materials and methods

The anti-inflammatory effect was evaluated through the mouse ear oedema test by means of two irritating agents, arachidonic acid (AA) and phorbol myristate acetate (PMA).

Results

Heliopsis longipes, affinin and isobutyl-decanamide displayed a marked anti-inflammatory effect on the AA model with ED₅₀ = 0.8, 1.2 and 0.9 mg/ear, respectively. Nimesulide (1 mg/ear) was used as a reference drug. In PMA model, the extract and two alkamides also showed a dose-dependent anti-inflammatory effect with ED₅₀ = 2.0, 1.3 and 1.1 mg/ear, respectively. Indomethacin (3 mg/ear) was used as reference drug.

Conclusions

These results could represent an important contribution to explain the anti-inflammatory ethnobotanical effects reported for Heliopsis longipes and other species containing affinin (spilanthol). For the first time the topical anti-inflammatory effects of Heliopsis longipes, affinin and isobutyl-decanamide were studied.     

Walnut leaf extract inhibits PTP1B and enhances glucose-uptake in vitro

Ethnopharmacological relevance

Walnut, Juglans regia L. (Juglandaceae), is one of the medicinal plants used to treat diabetic symptoms in Austrian folk medicine. The air-dried green leaves are either used as aqueous decoctions or liquor preparations and are consumed on a daily basis. We investigated the hypoglycemic effect of a methanolic Juglans regia leaf extract on glucose uptake, protein tyrosine phosphatase 1B (PTP1B) inhibition and peroxisome proliferator-activated receptor gamma (PPARγ) activation.

Material and methods

Hypoglycemic activity was assessed by glucose-uptake in C2C12 myocytes, inhibition of PTP1B and activation of PPARγ. Phytochemical characterization of the extract was carried out by LC–MS and GC–MS.

Results

Methanolic Juglans regia leaf extract enhanced the glucose uptake rate in C2C12 myocytes at concentrations of 25 µg/mL compared to untreated cells. This activity may partly be explained by the inhibition of PTP1B but not PPARγ agonism. LC–MS analyses revealed chlorogenic acid (1), 3-p-coumaroylquinic acid (2), a trihydroxynaphthalene-hexoside (3), as well as eight flavonoids (4–11) as main phenolic constituents in the active extract.

Conclusions

The finding that Juglans regia leaf extract enhances glucose uptake and inhibits PTP1B provides an in vitro-based rationale for the traditional use of walnut leaf preparations against elevated blood-glucose levels.     

In vitro antitrypanosomal and antileishmanial activity of plants used in Benin in traditional medicine and bio-guided fractionation of the most active extract

Ethnopharmacological relevance

The aim of the study was to evaluate the in vitro antitrypanosomal and antileishmanial activity of crude extracts of 10 plant species traditionally used in Benin to treat parasitic infections.

Materials and methods

For each species, dichloromethane, methanol and aqueous extracts were tested. Their antitrypanosomal and antileishmanial activities were evaluated in vitro on Trypanosoma brucei brucei (strain 427) (Tbb) and on promastigotes of Leishmania mexicana mexicana (MHOM/BZ/84/BEL46) (Lmm).

Results

The best growth inhibition was observed with the dichloromethane extracts of aerial parts of Acanthospermum hispidum DC. (Asteraceae) (IC₅₀ = 14.5 μg/ml on Tbb and 11.1 μg/ml on Lmm), twigs of Keetia leucantha (K. Krause) Bridson (syn. Plectronia leucantha Krause) (IC₅₀ = 5.8 μg/ml on Tbb), aerial parts of Byrsocarpus coccineus Schumach. & Thonn (syn. Rourea coccinea (Schumach. & Thonn.) Hook.f.) (IC₅₀ = 14.7 μg/ml on Tbb) and aerial parts of Carpolobia lutea G.Don. (IC₅₀ = 18.3 μg/ml on Tbb). All these extracts had a low cytotoxicity. It is not the case for the methanolic and water extracts of roots of Anchomanes difformis (Blume) Engl. (IC₅₀ = 14.7 and 13.8 μg/ml on Tbb) which were toxic at the same concentration range on WI38, human cells. A bio-guided fractionation of the most active extract of Keetia leucantha allowed to identify oleanolic acid and ursolic acid as responsible for the observed activities.

Conclusion

Our study gives some justification for antiparasitic activity of some investigated plants.     

Antiproliferative and antimetastatic effects of the ethanolic extract of Phellinus igniarius (Linnearus: Fries) Quelet

AIM OF THIS STUDY: Phellinus igniarius (Linnearus: Fries) Quelet (Phellinus igniarius) has been used in oriental countries for treatment of various diseases including cancer. However, it is unclear how Phellinus igniarius exerts anticancer effects. MATERIALS AND METHODS: In this study the ethanolic extract from the fruiting body of Phellinus igniarius (EEPI) was used to evaluate the antiproliferative and antimetastatic effects in human hepatocarcinoma SK-Hep-1 cells and rat heart vascular endothelial cells (RHE cells). RESULTS: We found that EEPI inhibited the proliferation of both cell lines in a dose-dependent manner, and the IC50 values at 48 h were 72 and 103 microg/ml for SK-Hep-1 cells and RHE cells, respectively. EEPI at non- or sub-cytotoxic concentrations (25-100 microg/ml) markedly inhibited the migration and invasion of SK-Hep-1 cells. EEPI added at 25 microg/ml significantly decreased the secretion of matrix metalloproteinase-2 (MMP-2) (49%, p<0.01) and vascular endothelial growth factor (VEGF) (13%, p<0.05) in SK-Hep-1 cells. EEPI at 25 microg/ml completely inhibited matrigel-induced tube formation in RHE cells. Importantly, EEPI (25 or 50 microg/ml) in combination with oxaliplatin (Oxa) or 5-flurouracil (5-FU) synergistically inhibited the proliferation of SK-Hep-1 cells. CONCLUSION: These results demonstrate the antiproliferative and antimetastatic effects of EEPI in vitro and the potential of EEPI as an adjuvant for chemotherapy.     

In vitro and in vivo anti-diabetic activity of Swertia kouitchensis extract

Ethnopharmacological relevance

Swertia kouitchensis has long been used as a folk medicine to treat hepatitis and diabetes in central-western China. Therefore, this study was aimed to evaluate the anti-diabetic activity of the plant ethanol extract.

Materials and methods

Firstly, the extract was tested for its inhibitory activity on α-amylase and α-glucosidase in vitro. Following that, insulin secretion test in NIT-1cell was performed. Then, oral sucrose or starch tolerance test of the extract were carried out in normal mice. After that, acute effect of the extract was executed in normal and streptozotocin-induced (60 mg/kg) diabetic mice. Eventually, long term effect of the extract was performed in diabetic mice for 4 weeks. Oral glucose tolerance test and biochemical parameters were estimated at the end of the study.

Results

Swertia kouitchensis extract could remarkably inhibit the activity of α-amylase and α-glucosidase and stimulate insulin secretion in vitro. And also the extract displayed anti-hyperglycemic activity, improved antioxidant capacity, ameliorated the hyperlipidemia and carbohydrate metabolism in diabetic mice.

Conclusions

Swertia kouitchensis exhibits considerable anti-diabetic activity and metabolic alterations in diabetic mice. These results provide a rationale for the use of Swertia kouitchensis to treat diabetes mellitus.     

Antidiabetic and antioxidant effects of Annona muricata (Annonaceae), aqueous extract on streptozotocin-induced diabetic rats

Ethnopharmacological relevance

The leaves of Annona muricata are used in Cameroon to manage diabetes and its complications. The aim of this study was to evaluate the antidiabetic, antioxidant activities and the potential toxicity of aqueous extract of Annona muricata in streptozotocin-induced diabetic rats.

Material and methods

Oral administration of Annona muricata aqueous extract (100 mg/kg or 200 mg/kg) was studied in normal and streptozotocin-induced diabetic rats. In long term treatment, 2 weeks after streptozotocin-induced diabetic rats, animals received plant extract during 28 consecutive days. For a protective effect, extract was administered 3 days prior to streptozotocin exposure and animals were observed 2 weeks without treatment.

Results

The plant extract was not effective in normal rats. In diabetic rats, single administration of the extract significantly reduced blood glucose levels by 75% and 58.22% respectively at the dose of 100 mg/kg and 200 mg/kg as compared to the initial value. Treatment of normal rats 3 days prior to diabetes induction showed that, Annona muricata extract has no effect within 72 h following STZ injection. However, after 14 days post-treatment, the extract at the dose of 100 mg/kg significantly reduced blood glucose levels as compared with initial value and diabetic control rats. Immunohistochemical staining of pancreatic β-cells of diabetic rats treated with the dose of 100 mg/kg expressed strong staining for β-cell compared to diabetic control. In a long-term study daily administration of Annona muricata aqueous extract for 28 days to diabetic rats, reduced blood glucose levels, serum creatinine, MDA, AST, ALT activity, and nitrite levels LDL-cholesterol. Total cholesterol, triglycerides, SOD, and CAT activity contents were restored.

Conclusion

These different results show that the antidiabetic activity of Annona muricata aqueous extract can be explained by its hypolipidaemic effect, its antioxidant and protective action on pancreatic β-cells, which in turn improve glucose metabolism.     

Antidiabetic activity of Ficus amplissima Smith. bark extract in streptozotocin induced diabetic rats

Ethnopharmacological relevance

Ficus amplissima commonly known as kal-itchchi have a long history of use in Ayurveda, Siddha and Unani herbal preparations in Indian traditional system of medicine. It has been used in folklore medicine for the treatment of diabetes.

Aim

The aim of the present study was to evaluate the effect of methanolic extract of the bark of Ficus amplissima (FAB) in streptozotocin-induced diabetic rats.

Materials and methods

Oral administration of FAB bark at the doses of 50 mg/kg, 100 mg/kg and 150 mg/kg was studied in normal, glucose-loaded and STZ—induced diabetic rats.

Results

The three doses caused significant reduction in blood glucose levels in all the models. The effect was more pronounced in 50 mg/kg and 100 mg/kg than 150 mg/kg. FAB also showed significant increase in serum insulin and body weight. The glycogen content in liver, skeletal muscle, total protein contents were markedly increased and marker enzymes of hepatic function of STZ-induced diabetic rats while there was significant reduction in the levels of serum triglyceride and total cholesterol. FAB also showed significant anti-lipid peroxidative effect in the pancreas of STZ-induced diabetic rats. The anti-diabetic effect of FAB was compared with glibenclamide, a well known hypoglycemic drug. Histological analysis showed the regenerative effect of FAB on the β-cells of diabetic rats.

Conclusion

Results of this experimental study indicated that FAB possessed anti-diabetic and antioxidant activities. Hence it could be used as a natural source of antidiabetic (Type-I) and antioxidant drug.     

Anti-tumor and pro-apoptotic activity of ethanolic extract and its various fractions from Polytrichum commune L.ex Hedw in L1210 cells

Ethnopharmacological relevance

Polytrichum commune L.ex Hedw is a traditional Chinese herb for treatment of fever, hemostatic, uterine prolapse and especially for lymphocytic leukemia, but the antitumor effect and its potential mechanism remains unclear.

Aims of the study

The present study was to investigate the possible anti-proliferative activity of ethanolic extract and the organic fractions from P. commune on murine leukemia L1210 cells.

Materials and methods

The content of ethanolic extract and its fractions was performed on HPLC analysis with gradient elution. L1210 cells were treated with different concentrations of ethanolic extract and its fractions at different time intervals. Cell viability was evaluated using MTT assay. Apoptotic cell death was monitored by nuclear condensation and confirmed by exposure of phosphatidylserine to outer leaflet of plasma membrane. Loss of mitochondrial membrane potential was analyzed by flow cytometry using rhodamine 123 staining.

Results

The obtained results showed that the cell viability of L1210 cells was reduced by ethanolic extract of P. commune in a concentration-dependent manner, and the IC₅₀ value was about 77.22 μg/ml at 24 h post treatment. The ethylacetate fraction displayed higher anti-tumor effect than that of chloroform and butanol fractions with 32.29 μg/ml (IC₅₀ value, 48 h). Microscopy studies revealed that ethanolic extract and ethylacetate fraction treated cells showed morphological characteristics of apoptosis such as chromatin condensation and DNA aggregation. Further, Annexin V-PE/ 7-AAD double staining showing the out leaflet of phosphatidylserine and the decline of mitochondrial membrane potential by flow cytometry confirmed that the extracts do, in fact, induce apoptosis in L1210 cells.

Conclusion

This is the first report on anti-tumor and pro-apoptotic effect of P. commune in cultured leukemia cells, which provides scientific basis for its usefulness as traditional medicine. Further studies are needed to confirm the precise mechanism not only the crude extract but as well the monomeric chemical substances of Polytrichum commune L.ex Hedw.     

Toxicity and anti-trypanosomal effects of ethanolic extract of Butyrospermum paradoxum (Sapotaceae) stem bark in rats infected with Trypanosoma brucei and Trypanosoma congolense

The ethanolic extract of Butyrospermum paradoxum stem bark, commonly used in the traditional treatment of various diseases including animal and human trypanosomosis in north-eastern Nigeria, was tested for toxicity and anti-trypanosomal efficacy in rats infected with Trypanosoma congolense and Trypanosoma brucei. Following intra-peritoneal administration, the extract induced behavioural changes, morbidity and mortality in the rats. The symptoms observed included anorexia, dehydration, depression, prostration, coma and death. These symptoms were noted at high doses (> 800 mg/kg) only. At necropsy, the pathological lesions were mainly congestion and oedema of the lungs, bronchi, bronchioles and the kidney, hepatomegally and focal necrosis of the liver cells. The severity of the symptoms and lesions were dose related. The intra-peritoneal LD50 of the extract was 820 mg/kg. The extract produced anti-trypanosomal effect through the complete suppression or delay in parasite establishment with reduction in the level of parasitaemia and the severity of the attendant disease as well as enhanced survival of the rats infected with Trypanosoma congolense and Trypanosoma brucei. The results suggest that the folkloric medicinal application of the extracts of Butyrospermum paradoxum has a pharmacological basis.