Neuroprotective effect of Alpinia galanga (L.) fractions on Aβ(25-35) induced amnesia in mice

Ethnopharmacological relevance

The rhizomes of Alpinia galanga (L.) Willd (Zingiberaceae), a ginger substitute for flavouring food was traditionally used as nervine tonic and stimulant.

Aim of the study

This investigation is designed to screen cognitive improvement of Alpinia galanga (AG) fractions in Alzheimer's type of amnesia in mice induced by Aβ_(25-35).

Materials and methods

Alzheimer's disease induced mice treated with fractions (n-hexane, chloroform and ethyl acetate) of AG in 200 and 400 mg/kg. Neurotoxicity was induced by intracerebroventricular injection of Aβ_(25-35) on the 14th day of 21 days drug treatment. Open field and water maze were carried to determine habituation memory and hippocampal memory. Na⁺/K⁺-ATPase, acetylcholinesterase (AChE) and antioxidant enzymes (SOD, GPx, catalase and vitamin C) were determined in brain tissue homogenate to estimate the brain biochemical changes and its anti-amnesic potential with intensity of oxidative stress signaling. Further bioactive (chloroform) fraction was eluted through column chromatography to identify the lead molecules.

Results

Increased habituation memory and decreased escape latency in behavioral parameter are the indicative of the cognitive enhancement after treatment with Alpinia galanga fractions. Increment in Na⁺/K⁺-ATPase and antioxidant activity depicts brain membrane integrity improvement and free radical scavenging property. AChE level was decreased to improve the cognition by enhancing cholinergic transmission.

Conclusion

Anti-amnesic effect was exerted by various fractions of Alpinia galanga. Among all fractions, preeminent neuroprotection was exerted by chloroform fraction, which has compound, 1′δ-1′-acetoxyeugenol acetate and it may be a potential therapeutic agent for Alzheimer's type of amnesia. These results further motivate us to explore the activity of lead compound's anti-amnesic effect on transgenic mice model of AD.     

Typhonium flagelliforme induces apoptosis in CEMss cells via activation of caspase-9, PARP cleavage and cytochrome c release: Its activation coupled with G0/G1 phase cell cycle arrest

Ethnopharmacological relevance

The plant Typhonium flagelliforme (TF), commonly known as ‘rodent tuber’ in Malaysia, is often used as traditional remedy for cancer, including leukemia.

Aim of the study

We had previously identified morphologically that the linoleic acid rich fraction (DCM/F7) from the tubers of this plant induces selective anti-proliferative effects and apoptosis in CEMss cells. In this present study, we subjected the same DCM/F7 fraction to cell based activity analyses in order to determine the possible mechanism of cell death in leukemic CEMss cells in vitro.

Materials and methods

Extraction of Typhonium flagelliforme tuber has done and fractionation has been done by vacuum liquid column chromatography. The anti-proliferative activity was assayed using MTT and the apoptosis detection was done by Annexin V and DNA laddering assay. Colorimetric caspase assay and immunoblot analysis were employed to detect the expression of protein associated with cell death. Cell cycle analysis was done using flow cytometry.

Results

We found that the cancer inhibitory effect of the DCM/F7 fraction in CEMss cells was 3 ± 0.08 μg/ml (IC₅₀). An early apoptotic induction in CEMss cells was observed by Annexin V assay, which showed a clear dose-dependent DNA fragmentation being observed in gel electrophoresis at 10 and 20 μg/ml. The DCM/F7 fraction at 3 μg/ml significantly arrested CEMss cells at G0/G1 phase (p < 0.05). A constant but increasing pattern-related Sub-G0/G1 index was observed between 12 and 72 h treatment. In relation to this, we further investigated the biochemical events leading to cell death and found that the DCM/F7 fraction increased the cellular levels of caspase-3 and -9 on treated cells. Our results indicated that cytochrome c from mitochondria into the cytosol increased gradually as the DCM/F7 concentration increases, which later lead to the subsequent cleavage of PARP in to 85 kDa fragments. On the contrary, Bcl-2 protein was found to decrease concomitantly during treatment.

Conclusions

Collectively, results presented in this study demonstrated that the DCM/F7 fraction inhibited the proliferation of leukemia cells, leading to the programmed cell death, which was confirmed to be through the mitochondrial pathway.     

Bioactivity-guided fractionation for anti-fatigue property of Acanthopanax senticosus

Ethnopharmacological relevance

The root of Acanthopanax senticosus (also called Eleutherococcus senticosus or Siberian ginseng) has been used extensively in China, Russia and Japan as an adaptogen to fight against stress and fatigue.

Aim of the study

The present study was designed to ascertain the anti-fatigue property of Acanthopanax senticosus by load-weighted swimming test, sleep deprivation test, also to isolate and characterize the active constituents.

Materials and methods

Animals were orally administered with the extract of Acanthopanax senticosus. The anti-fatigue effects of the four fractions with different polarities from the 80% ethanol extract, and the different eluates collected from D101 macroporous resin chromatography and eleutheroside E, were examined based on the weight-loaded swimming capacity (physical fatigue) and the change of biochemical parameters in ICR mice. Moreover, the active fraction was later submitted to sleep-deprived mice (mental fatigue).

Results

The results shown that the n-butanol fraction significant extends the swimming time of mice to exhaustion. Furthermore, the 60% ethanol-water eluate, more purified eleutherosides (including eleutheroside E, E₂ and derivatives), were the exactly active constituents. Two compounds were isolated, which were identified as eleutheroside E, E₂.

Conclusions

The eleutherosides possess the potent abilities to alleviate fatigue both in physical and mental fatigue. Eleutheroside E may be responsible for the pharmacological effect of anti-fatigue. Furthermore, the possible mechanisms were reduced the level of TG by increasing fat utilization, delayed the accumulation of blood urea nitrogen (BUN), and increased the LDH to reduce the accumulation of lactic acid in muscle and then protect the muscle tissue.     

Protection against 1-methyl-4-phenylpyridinium ion (MPP+)-induced apoptosis by water extract of ginseng (Panax ginseng C.A. Meyer) in SH-SY5Y cells

Ethnopharmacological relevance

The present study investigates the protective effects of water extract of ginseng (Panax ginseng C.A. Meyer) against 1-methyl-4-phenylpyridinium ion (MPP⁺)-induced cytotoxicity in SH-SY5Y human neuroblastoma cells and explores the underlying mechanisms. The approach may be used for screening therapeutic agents for degenerative disorders such as Parkinson's disease.

Materials and methods

SH-SY5Y human neuroblastoma cells were used to analyze the protective effects of water extract of ginseng (WEG) against multiple parameters such as MPP⁺-induced viability, oxidative injury, expression of Bax, Bcl-2, cytochrome c and cleaved caspase-3.

Results

WEG exerted inhibitory effect on cell death, overproduction of ROS, elevated Bax/Bcl-2 ratio, release of cytochrome c and activation of caspase-3 expression in MPP⁺-treated SH-SY5Y cells.

Conclusions

WEG exhibited significant protective effects against MPP⁺-induced cytotoxicity in SH-SY5Y cells possibly through the suppression of ROS generation and the inhibition of mitochondria-dependent apoptotic pathway.     

Neuroprotective effects of Eleutherococcus senticosus bark on transient global cerebral ischemia in rats

Ethnopharmacological relevance

Eleutherococcus senticosus Maxim., classified into the family of Araliaceae, is used in a variety of diseases in traditional Korean medicine including ischemic heart disease.

Aim of the study

To determine the neuroprotective effects of Eleutherococcus senticosus on global cerebral ischemia.

Materials and methods

A four-vessel occlusion (4-VO) rat model was used to evaluate the potential protective effects against transient global cerebral ischemia ethanol extracts of Eleutherococcus senticosus was orally administered at doses of 3, 30, and 300 mg/kg twice at times of 0 and 90 min after reperfusion. The effects on memory deficit were investigated by using a Y-maze neurobehavioral test after brain ischemia, and the effects on hippocampal neuronal damage were measured 7 days after ischemia. The expressions of glial fibrillary acid protein (GFAP), CD11b antibody (OX-42), and cyclooxygenase-2 (COX-2) were investigated by immunohistochemistry.

Results

Oral administration of Eleutherococcus seticosus at 30, 100 and 300 mg/kg significantly reduced hippocampal CA1 neuronal death by 3.5%, 25.9% and 53.1%, respectively, compared with a vehicle-treated group. Oral administration of Eleutherococcus senticosus at 300 mg/kg inhibited 81.9% of the decrease in spontaneous alternation induced by 4-VOin the Y-maze test, and also attenuated ischemia-induced activation of COX-2, GFAP and OX-42 in the hippocampal CA1 region.

Conclusion

Eleutherococcus senticosus protects delayed neuronal death in the CA1 region of the hippocampus against global cerebral ischemia in rats with the recovery of spatial memory, which can be considered as the normal functioning of the hippocampus. Regarding the immunohistochemical study, the effect of Eleutherococcus senticosus may be attributable to its anti-inflammatory properties through the inhibition of COX-2 expression, microglia and astrocyte expression.     

Antidepressant effect of Valeriana wallichii patchouli alcohol chemotype in mice: Behavioural and biochemical evidence

Ethnopharmacological relevance

Valeriana wallichii DC, an ayurvedic traditional medicine has now been shown to exist chemically as three distinct chemotypes. The study aimed to investigate the antidepressant effect of dichloromethane extract of Valeriana wallichii patchouli alcohol chemotype.

Materials and methods

Antidepressant effect of dichloromethane extract of Valeriana wallichii (10, 20 and 40 mg/kg, p.o.) using forced swim test, was determined in both acute and chronic study. The neurotransmitter levels were estimated in mouse forebrain after two weeks of dosing.

Results

Single administration of extract (40 mg/kg) significantly inhibited the immobility period in mice (p < 0.05). Similarly, chronic administration of extract (20 and 40 mg/kg) significantly reduced the immobility period and significantly increased the levels of norepinephrine and dopamine in mouse forebrain (p < 0.05).

Conclusions

The extract demonstrated antidepressant effect and significantly increased the norepinephrine and dopamine levels in forebrain.     

In vitro growth stimulatory and in vivo wound healing studies on cycloartane-type saponins of Astragalus genus

Aim of the study

The present study was undertaken to evaluate the wound healing effects of the four chief saponins of Astragalus species [cycloastragenol (CA), astragaloside IV (AG), cyclocephaloside I (CCI) and cyclocanthoside E (CCE)].

Material and methods

Effects of cell viability and proliferation of the isolated compounds were evaluated by the MTT assay on human keratinocyte. The wound healing activity was studied by using in vitro wound healing, proliferation and migration scratch assay. In order to see in vivo effectiveness of the compounds, an animal study with Sprague-Dawley male rats at the age of 12 weeks was carried out, and then the main histological outcomes were investigated to observe reepithelization, neovascularization, and presence of inflammatory cells, granulation tissue amount and maturation.

Results

All the compounds increased both fibroblast proliferation and migration, but the effects were much superior for CA at 1 ng/ml concentration. Among the compounds, based on the histological findings, 5% CA preparation was found to be the most remarkable in vivo wound healing agent showing greater cell density, more regularly organized dermis and more newly formed blood vessels.

Conclusion

Results of this study indicate that the cycloartane-type saponins are the principal constituents responsible for wound healing activities of the roots of Astragalus species substantiating its use in traditional medicine.     

The ethyl acetate extract of PGP (Phellinus linteus grown on Panax ginseng) suppresses B16F10 melanoma cell proliferation through inducing cellular differentiation and apoptosis

Ethnopharmacological relevance

Phellinus linteus and Panax ginseng have been widely used as traditional herbal medicines to treat various diseases including cancer in East Asia.

Aim of the study

The present study sought to investigate the possible mechanism in anti-proliferative effect of Phellinus linteus that was grown on Panax ginseng (PGP) on B16F10 melanoma cells.

Materials and methods

The anti-proliferative effect of PGP on B16F10 was evaluated by CCK-8 assays. Apoptotic cells were detected by flow cytometry analysis. The proteins involved in apoptosis and cellular differentiation were assessed by immunoblot analysis. Ginsenosides contents of PG or PGP were analyzed using HPLC.

Results

The ethyl acetate fraction (EtOAc) of PGP exhibited the strongest anti-proliferative activity among PGP fractions (butanol or water) on B16F10 cells. PGP EtOAc extract showed stronger inhibitory effect than Panax ginseng (PG) or Phellinus linteus (PL) EtOAc extract on B16F10 melanoma cell proliferation. PGP EtOAc extract induced the dendrite-like structures and the melanin production in B16F10 cells. PGP EtOAc extract increased a sub-G1 cell population through inducing p53/p21 and activated caspase-8 protein expression in B16F10 cells. Notably, PGP EtOAc extract contained ginsenosides Rd, Rg3, Rb2, Rg1 and Rb1 more than PG EtOAc extract. Rd and Rg3 significantly inhibited B16F10 cell proliferation.

Conclusion

Our data suggest that PGP EtOAc extract inhibits B16F10 cell proliferation through inducing apoptosis and cellular differentiation.     

Effect of Emblica officinalis (fruit) against UVB-induced photo-aging in human skin fibroblasts

Ethnopharmacological relevance

Emblica officinalis fruit (EO), commonly known as Amla is a reputed traditional medicine and functional food used in Indian subcontinent. It has long been used in Indian folk medicine to treat liver diseases, stomach ulcers, inflammatory diseases, metabolic disorders, geriatric complaints, skin disorders and beauty care.

Aim of the study

Recently, it has been shown to promote pro-collagen content and inhibit matrix metalloproteinase levels in skin fibroblast. The aim of the present study was to investigate the efficacy of EO to inhibit UVB-induced photo-aging in human skin fibroblasts.

Materials and methods

Mitochondrial activity of human skin fibroblasts was measured by MTT-assay. Quantifications of pro-collagen 1 and matrix metalloproteinase 1 (MMP-1) release were performed by immunoassay techniques. Hyaluronidase inhibition assay was studied in vitro using bovine testicular hyaluronidase and human umbilical cord hyaluronic acid. Cell cycle analysis was performed by flowcytometry using propidium iodide.

Results

EO stimulated, the otherwise UVB inhibited cellular proliferation and protected pro-collagen 1 against UVB-induced depletion via inhibition of UVB-induced MMP-1 in skin fibroblasts (10-40 μg/mL, p > 0.001). EO exhibited inhibitory activity of hyaluronidase (10-40 μg/mL, p > 0.001). Treatment with EO also prevented UVB disturbed cell cycle to normal phase.

Conclusion

The results of the present study suggests that EO effectively inhibits UVB-induced photo-aging in human skin fibroblast via its strong ROS scavenging ability and its therapeutic and cosmetic applications remain to be explored.     

Indigofera suffruticosa Mill as new source of healing agent: involvement of prostaglandin and mucus and heat shock proteins

Ethnopharmacological relevance

Indigofera suffruticosa is specie typical of the “Cerrado” or Brazilian savannah; it is a member of the Fabaceae family - in folkmedicine is used for gastric disorders, infection and inflammation.

Aim of the study

Ethyl acetate fraction (AcF) and aqueous fraction (AqF) of the methanolic extract of I. suffruticosa leaves were evaluated against acute gastric ulcer. The AcF fraction was selected to assess its activity in ulcer healing and its gastroprotective effects via mucus and gastric secretion.

Materials and methods

The gastroprotective action of AcF and AqF fractions were evaluated in a rodent experimental model. The action mechanisms, involvements of the antisecretory action, mucus and prostaglandin production, toxicological and healing activity of the AcF (100 mg/kg, p.o.) were evaluated. We also used histological analysis (HE and PAS) and immunohistochemical (PCNA and HSP-70) assays to evaluate the effects of I. suffruticosa.

Results

AcF significantly inhibited the gastric mucosal damage caused by ethanol. This effect was statistically significant in 100 mg/kg group compared vehicle. AcF did not interfered with gastric secretion, significantly increased the PGE₂ and mucus production (validated in PAS technique). The gastroprotection was attenuated by pretreatment with N-ethylmaleimide, but not L-NAME. In acid-acetic-induced ulcer model AcF accelerated ulcer healing. Immunohistochemistry analysis showed induction of proliferating cell (PCNA) and heat shock protein (HSP 70).

Conclusions

These results showed that AcF acted as gastroprotective agent stimulating prostaglandin, mucus and HSP70.     

Intracellular replication inhibitory effects of Galla Rhois ethanol extract for Brucella abortus infection

Ethnopharmacological relevance

Galla Rhois (GR) has long been applied in traditional Korean and Oriental medicine. Although GR has an anti-bacterial effect, the anti-bacterial mechanism and therapeutic efficiency of GR for intracellular parasitic Brucella infection are still unclear.

Aim of the study

The objective of this study was to investigate the antibacterial and therapeutic effects of GR ethanol extract (GRE), which is a natural antibacterial component for the treatment of Brucella abortus infection.

Materials and methods

The antibacterial activity of GRE towards Brucella abortus was evaluated by incubating Brucella abortus with GRE. Following treatment with GRE, Brucella abortus adherence, uptake, intracellular growth, and intracellular trafficking in macrophages were monitored. Mice were infected intraperitoneally with Brucella abortus and treated orally with GRE for 14 days, and then the weight and CFUs from each spleen were monitored.

Results

The viability of Brucella abortus was markedly decreased in a dose-dependent manner. Moreover, Brucella abortus internalization and intracellular growth within macrophages were reduced in GRE-treated cells. The number of bacteria that adhered to GRE-pretreated cells was significantly lower than that of untreated cells. With regards to intracellular trafficking, treatment with GRE augmented the colocalization of Brucella abortus-containing phagosomes with LAMP-1. GRE-treated mice showed considerably decreased weight and bacterial burdens in the spleen compared to untreated mice.

Conclusion

GRE exhibits antibacterial and protective effects on Brucella abortus in vitro and in vivo. These results highlight the beneficial effects of GRE in the prevention and treatment of brucellosis.     

In vivo anti-tussive activity and structural features of a polysaccharide fraction from water extracted Withania somnifera

Aim of the study

Anti-tussive drugs are amongst the most widely used medications worldwide; however no new class of drugs has been introduced into the market for many years. The present study aims at evaluating the structural features and in vivo anti-tussive activity of a polysaccharide fraction from water extracted Withania somnifera.

Materials and methods

Herein, we have analyzed water extracted material of Withania somnifera using chemical, chromatographic, spectroscopic and biological methods.

Results

A polysaccharide fraction (F3) containing arabinosyl, galactosyl and galacturonosyl residues were obtained by anion exchange chromatography of the water extracted material. This polymer is branched and contained (1,5)-/(1,3,5)-linked arabinofuranosyl, (1,3)-/(1,6)-/(1,3,6)-linked galactopyranosyl residues together with small amount of terminal rhamnopyranosyl and terminal arabinofuranosyl residues. Peroral administration of this pectic arabinogalactan in a dose of 50 mg kg⁻¹ body weight (b.w.) decreased the number of cough efforts induced by citric acid in guinea pigs like that of codeine.

Conclusions

This study provides a scientific basis for the past and present ethnomedical uses of this plant.     

Inhibition of 2,4-dinitrofluorobenzene-induced atopic dermatitis by topical application of the butanol extract of Cordyceps bassiana in NC/Nga mice

Ethnopharmacological significance

The Cordyceps species are insect-borne mushrooms that have been ethnopharmacologically used for skin diseases such as eczema and dermatitis.

Aim of the study

In this study, we investigated the curative effects of the butanol fraction (CBBF) of Cordyceps bassiana on atopic dermatitis.

Materials and methods

Dermatitis was induced by repeated application of 2,4-dinitrofluorobenzene (DNFB) in NC/Nga mice. After a topical application of CBBF on the skin lesions, the dermatitis score, epidermal thickness, mast cell number, and interleukin (IL)-4 and interferon (IFN)-γ, as well as the levels of histamine and immunoglobulin E (IgE) in the serum, were measured. Moreover, effect of CBBF on histamine release was examined using RBL-2H3 under stimulation with 2,4-dinitrophenylated bovine serum albumin (DNP-BSA).

Results

CBBF inhibited atopic dermatitis symptoms and signs in the DNFB-treated NC/Nga mice. The suppressive activity of topically applied CBBF may be due to the dose-dependent blockade of a series of immunopathological events, including the release of histamine, the production of IgE, and the secretion of IL-4 and IFN-γ. However, this extract did not directly suppress the degranulation process, assessed by measuring β-hexosaminidase release.

Conclusions

Our results suggest that CBBF can be applied as an effective herbal remedy to treat atopic dermatitis.     

Anti-oxidation activity of the crude polysaccharides isolated from Polygonum Cillinerve (Nakai) Ohwi in immunosuppressed mice

Aim of the study

Polygonum Cillinerve (Nakai) Ohwi is commonly used in China for over 2000 years. Previous research has shown that the crude polysaccharides extracted from Polygonum Cillinerve (Nakai) Ohwi (PCCP) have the scavenging free radicals and anti-tumor activities in vitro. In present study, PCCP were further approached the perspective of their anti-oxidation in immunosuppressed mice.

Methods and materials

ICR mice were treated firstly with cyclophosphamide (CY, 150 mg/kg), 1 day later, treated with different dosages of PCCP or saline solution once daily for 21 days. Twenty-four hours later for the last drug administration, the animals were weighed, and then killed by decapitation. The liver, spleen, and thymus indices were investigated, and the biochemical parameters were evaluated for various tissues (liver, heart, and kidney).

Results

The administration of PCCP with gavage was able to overcome the cyclophosphamide-induced immunosuppression, and significantly to raise the TOC, CAT, SOD, and GSH-Px level. It also raised the liver, spleen, and thymus indices, and decreased the MDA level in mice.

Conclusions

PCCP possess the pronounced free radical-scavenging and antioxidant activities, and could play an important role in the prevention of oxidative damage in immunological system.     

Evaluation of the effects of Astragalus mongholicus Bunge saponin extract on central nervous system functions

Ethnopharmacological relevance

In traditional medicine, Astragalus mongholicus (AM) has been used for the treatment of general weakness, chronic illness, and to increase overall vitality.

Aim of the study

The present study investigated possible effects of the saponin fraction of AM on the central nervous system. Moreover, its effects on locomotor activity, anxiety, and hippocampal morphology were studied.

Material and methods

AM extract was tested for its effects on locomotor activity using the Moti-Test, for situational anxiety in the elevated plus maze, and for anticonvulsant activity against acute pentylenetetrazole (PTZ)-induced seizures and in the PTZ kindling model.

Results

It was shown that AM (50, 100, 200 mg/kg) did not interfere with locomotor activity and situational anxiety as measured in the elevated plus maze. In these doses, AM significantly suppressed pentylenetetrazole (PTZ)-induced seizures (p < 0.05). Its anticonvulsant efficacy was also evident against repeated PTZ seizures (p < 0.05). This suggests potential therapeutic usefulness. After subchronic application, the number of cells in hippocampal CA1 was reduced, whilst the cell number in CA3 and hilus remained unaffected.

Conclusions

Doses of AM extract which did not interfere with locomotor acticity and situational anxiety appear to be useful in the treatment of convulsive disorders. The mechanisms underlying this effect on hippocampal morphology are not yet understood.     

Identification of hypoglycaemic compounds from berries of Juniperus oxycedrus subsp. oxycedrus through bioactivity guided isolation technique

Ethnopharmacological relevance

Decoction of Juniperus oxycedrus subsp. oxycedrus L. (Cupressaceae) berries is used internally as tea and pounded fruits are consumed to lower blood glucose levels in Turkey.

Aim of the study

To evaluate hypoglycaemic and antidiabetic activity of J. oxycedrus subsp. oxycedrus berries and to identify active compounds through bioactivity guided isolation technique.

Material and methods

Hypoglycaemic effect of J. oxycedrus subsp. oxycedrus (Joso) berry extracts on oral administration was studied using in vivo models in normal, glucose-hyperglycaemic rats. Streptozotocin induced diabetic rats were used to examine antidiabetic activity of Joso extracts, subextracts, fractions, subfractions and shikimic acid (SA).

Results

Through in vivo bioactivity-guided fractionation processes, shikimic acid, 4-O-β-d-glucopyranosyl ferulic acid and oleuropeic acid-8-O-β-d-glucopyranoside were isolated from the n-butanol subextract by silica gel and reverse phase column chromatography as the main active ingredient of the active subfraction. After 8 days administration of the major compound shikimic acid, blood glucose levels (24%), malondialdehyde levels in kidney tissues (63-64%) and liver enzymes (AST, ALT, ALP) of diabetic rats were decreased.

Conclusion

Results indicated that Joso berry extract and its active constituents might be beneficial for diabetes and its complications.     

Antisecretory actions of Baccharis trimera (Less.) DC aqueous extract and isolated compounds: analysis of underlying mechanisms

Ethnopharmacological relevance

Baccharis trimera (Less.) DC. (Asteraceae) is a species native to South America used in Brazilian folk medicine to treat gastrointestinal and liver diseases, kidney disorders and diabetes. Previous studies from this laboratory confirmed the antacid and antiulcer activities of the plant aqueous extract (AE) in rat and mouse models.

Aim of the study

To investigate the mechanisms involved in the antacid action of AE and isolated compounds from Baccharis trimera.

Materials and methods

AE was assayed in vivo in cold-restraint stress gastric ulcers and in pylorus-ligated mice. Nine fractions (F2-F10) previously isolated from AE were assayed in vitro on acid secretion measured as [¹⁴C]-aminopyrine ([¹⁴C]-AP) accumulation in rabbit gastric glands, and on gastric microsomal H⁺, K⁺-ATPase preparations. Chlorogenic acids (F2, F3, F6, F7), flavonoids (F9), an ent-clerodane diterpene (F8) and a dilactonic neo-clerodane diterpene (F10) have been identified in these fractions.

Results

Intraduodenal injection of AE (1.0 and 2.0 g/kg) in 4 h pylorus-ligated mice decreased the volume (20 and 50%) and total acidity (34 and 50%) of acid secretion compared to control values. Administered orally at the same doses AE protected against gastric mucosal lesions induced in mice by restraint at 4 °C. Exposure of isolated rabbit gastric glands to fractions F8 (10-100 μM) and F9 (10-300 μg/ml) decreased the basal [¹⁴C]-AP uptake by 50 and 60% of control (Ratio = 6.2 ± 1.1), whereas the remaining fractions were inactive. In the presence of the secretagogues F2 and F4 (30-300 μg/ml) decreased the [¹⁴C]-AP uptake induced by histamine (His) with a 100-fold lower potency than that of ranitidine. F5 and F6 reduced the [¹⁴C]-AP uptake stimulated by carbachol (CCh), but they were 10 to 20-fold less potent than atropine. F8 (diterpene 2) and F9 (flavonoids) decreased both the His- and CCh-induced [¹⁴C]-AP uptake, whereas F10 (diterpene 1) was inactive against the [¹⁴C]-AP uptake stimulated by secretagogues. Diterpene 2 was the most active of all tested compounds being 7-fold less potent than ranitidine and equipotent to atropine in reducing acid secretion in vitro. This compound also reduced the gastric H⁺, K⁺-ATPase activity by 20% of control, while the remaining fractions were inactive on the proton pump in vitro.

Conclusions

The results indicate that Baccharis trimera presents constituents that inhibit gastric acid secretion by acting mainly on the cholinergic regulatory pathway. The plant extract also contains compounds that exert moderate inhibition of the histaminergic regulatory pathway of acid secretion and the gastric proton pump. Altogether these active constituents appear to provide effective inhibition of acid secretion in vivo, which may explain the reputed antiulcer activity of the plant extract.     

The potential role of female flowers inflorescence of Typha domingensis Pers. in wound management

Aim of the study

Female flowers inflorescence of Typha species including Typha domingensis Pers. are used externally for burns and wound healing in Turkish folk medicine. In order to verify the folkloric assertion, the female and male flowers inflorescences were individually submitted to in vivo wound models.

Material and methods

Ointment formulations prepared directly either from the male or female flowers inflorescences of Typha domingensis in 5% and 10% concentrations were submitted to activity testing. After that, female flowers inflorescence was further submitted to successive extractions with solvents in increasing polarity; i.e., n-hexane, chloroform, methanol and water and the wound healing activity of each extract was investigated. The linear incision and circular excision wound models were used for the evaluation of the healing potential of the test materials in rats and mice. Tissue sections were also evaluated by histopathological techniques.

Results

Remarkable wound healing activity was observed only for the female flowers inflorescence at 5% concentration in ointment base and its methanolic and aqueous extracts. The wound healing effect was found comparable to that of reference ointment Madecassol^®. The results of histopathological evaluation supported the outcome of both linear incision and circular excision wound models.

Conclusion

The experimental study revealed that the female flowers inflorescence of Typha domingensis displayed notable wound healing activity in mice and rats, at the models tested.     

Analgesic, anti-inflammatory and antipyretic activities of the petroleum ether fraction from the ethanol extract of Desmodium podocarpum

Ethnopharmacological relevance

Desmodium podocarpum is a plant that has been used in the folk medicine to treat febrile diseases, cough and bleeding wounds. However, there is no scientific basis or reports in the modern literature regarding its effectiveness as an analgesic, anti-inflammatory and antipyretic agent.

Aims of the study

The objective of this study is to evaluate the analgesic, anti-inflammatory and antipyretic activities of the petroleum ether fraction (PEF) from the ethanol extract of Desmodium podocarpum.

Materials and methods

PEF (50, 100, 200 mg/kg) was estimated for its pharmacological properties by using the acetic acid-induced writhing test, the hot plate test, the Carrageenan-induced rat paw edema model, the dimethylbenzene-induced mouse inflammation model, and the lipopolysaccharide (LPS)-induced rat fever model. In addition, the acute toxicity of PEF was also studied.

Results

PEF significantly and dose-dependently inhibited the writhing responses in mice, increased reaction time of mice in the hot plate test, reduced carrageenan-induced paw edema in rats and the dimethylbenzene-induced ear edema in mice, and attenuated LPS-induced fever in rats. No death of mice was observed when orally administered PEF up to 4.2 g/kg.

Conclusions

These findings suggest that PEF possesses evident analgesic, anti-inflammatory and antipyretic activities, and has a favorable safety, which supports the use of Desmodium podocarpum as an analgesic, anti-inflammatory and antipyretic drug in the folk medicine.