Protective effects of Gastrodia elata Blume on MPP-induced cytotoxicity in human dopaminergic SH-SY5Y cells

Aim of the study

Gastrodia elata (GE) Blume (Orchidaceae) has been traditionally used as a folk medicine in Oriental countries since centuries for their variety of therapeutic benefits. This study is an attempt to investigate the protective effects of GE extract against MPP⁺-induced cytotoxicity in human dopaminergic SH-SY5Y cells and explore the neuroprotective mechanisms involved.

Materials and methods

Human dopaminergic SH-SY5Y cells were used to demonstrate the protective effects of GE against multiple parameters such as MPP⁺-induced cell viability, oxidative damage, expression of Bcl-2 and Bax, caspase-3 and poly(ADP-ribose) polymerase proteolysis.

Results

GE effectively attenuated the cytotoxicity and improved cell viability in a dose-dependent manner. GE was effective in inhibiting both, the increased production of reactive oxygen species (ROS) and increase in Bax/Bcl-2 ratio, cleaved caspase-3 and PARP proteolysis.

Conclusion

Data from this study suggests the protective effects of GE on MPP⁺-induced cytotoxicity in dopaminergic cells, which may be ascribed to its significant anti-oxidative and anti-apoptotic properties, thus, GE might prove to be a valuable therapeutic agent for the treatment of various neurodegenerative diseases including progressive Parkinson's disease (PD).     

Chrysanthemum morifolium Ramat (CM) extract protects human neuroblastoma SH-SY5Y cells against MPP-induced cytotoxicity

Ethnopharmacological relevance

Chrysanthemum morifolium Ramat (Asteraceae) has (CM) long been used in Korean and Chinese traditional herbal medicines with numerous therapeutic applications.

Aim of the study

To evaluate the neuroprotective activities of Chrysanthemum morifolium (CM) extract against 1-methyl-4-phenylpridinium ions (MPP⁺), Parkinsonian toxin through oxidative stress and impaired energy metabolism, in human SH-SY5Y neuroblastoma cells and the underlying mechanisms.

Materials and methods

The effects of CM against MPP⁺-induced cytotoxicity and neuronal cell viability, oxidative damage, the expression of Bcl-2 and Bax, caspase-3 and poly(ADP-ribose) polymerase (PARP) proteolysis were evaluated by using SH-SY5Y neuroblastoma cells.

Results

CM effectively inhibited the cytotoxicity and improved cell viability. CM also attenuated the elevation of reactive oxygen species (ROS) level, increase in Bax/Bcl-2 ratio, cleavage of caspase-3 and PARP proteolysis.

Conclusion

These results demonstrate that CM possesses potent neuroprotective activity and therefore, might be a potential candidate in neurodegenerative diseases such as Parkinson's disease.     

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.     

Modulating effect of Leptadenia reticulata (Retz) Wight & arn against chromate (VI)-induced immunosuppression and oxidative stress on mouse splenic lymphocytes and bone marrow derived macrophages

Ethnopharmacological relevance

Leptadenia reticulata (Retz) Wight & arn is mentioned in the ancient ayurvedic literature as an immune booster and rejuvenator.

Aims of the study

To investigate, the effects of different forms of the extract of Leptadenia reticulata [Aqueous extract (JAE), Padavashesha kashaya (JPK) and Tarpana kashaya (JTK)] to alleviate the experimental immunosuppression induced by the immunotoxicant chromate (VI) in vitro.

Materials and methods

Standard cell proliferation and cytotoxicity assays like MTT assay, trypan blue dye exclusion test, neutral red dye uptake test, NBT reduction test, determination of percentage cell survival and estimation of markers of oxidative stress were performed in the study. The study was conducted on primary cultures of mouse splenic lymphocytes and bone marrow derived macrophages.

Results and Conclusions

Treatment with all the three forms of the extract used in the study offered protection against chromate (VI)-induced immunosuppression and the overall protective effect was found to be superior in the case of the aqueous extract of Leptadenia reticulata (JAE). These results confirm that Leptadenia reticulata acts as a modulator and alleviates the immunosuppressive conditions induced by chromate (VI).     

Hypotensive effect of aqueous extract of Averrhoa carambola L. (Oxalidaceae) in rats: an in vivo and in vitro approach

Aim of the study

Averrhoa carambola L. (Oxalidaceae) leaves are used in Brazilian traditional medicine to treat hypertension. This study was conducted to evaluate the hypotensive effect of the aqueous extract of Averrhoa carambola (AEAc) and its underlying mechanisms in the isolated rat aorta.

Materials and methods

The effect of AEAc on the mean arterial pressure (MAP) was determined in vivo in anesthetized rats. In vitro, thoracic aortic rings were isolated and suspended in organ baths, and the effects of AEAc were studied by means of isometric tension recording experiments. In HPLC analysis, the fingerprint chromatogram of AEAc was established.

Results

In normotensive rats, AEAc (12.5-50.0 mg/kg, i.v.) induced dose-dependent hypotension. In vitro, AEAc caused a depression in the E_max response to phenylephrine without a change in sensibility. Also, in a depolarized Ca²⁺-free medium, AEAc inhibited CaCl₂-induced contractions and caused a concentration-dependent rightward shift of the response curves, indicating that AEAc inhibited the contractile mechanisms involving extracellular Ca²⁺ influx.

Conclusions

These results demonstrate the hypotensive effects of AEAc, and these effects may, in part, be due to the inhibition of Ca²⁺, which supports previous claims of its traditional use.     

Apocynum venetum leaf aqueous extract inhibits voltage-gated sodium channels of mouse neuroblastoma N2A cells

Ethnopharmacological relevance

Apocynum venetum Linn. (Apocynaceae family), also called Luobuma, is a shrub which grows widely in the Xinjiang Autonomous Region of China. Its leaves are used in herbal tea for the treatment of hypertension, anxiety and depression. Animal studies have also shown that Apocynum venetum leaf extract (AVLE) also exerts anti-depressant and anti-anxiety activities. The effects of AVLE on neuronal tissues in vitro are not fully understood.

Materials and methods

Using the whole-cell voltage-clamp method, we studied the effects of AVLE on ion channels in cultured mouse neuroblastoma N2A cells.

Results

AVLE inhibited voltage-gated inward Na⁺ current in a reversible and concentration-dependent manner (half-inhibitory concentration was 18 μg/ml and maximum inhibition at 100 μg/ml). AVLE specifically promoted steady-state inactivation of Na⁺ channels but did not affect voltage-dependence of activation. The inhibitory effect was not use-dependent and was not affected by 300 μM L-NAME, suggesting that NO was not involved in the action of AVLE in neuronal cells. AVLE also had a mild inhibitory effect on voltage-gated K⁺ channels, but did not affect ATP-sensitive K⁺ channels.

Conclusions

Since voltage-gated Na⁺ and K⁺ channels are associated with neuronal excitability and therefore affect neurotransmission, the modulation of neuronal ion channels by AVLE may exert neuropharmacological effects. In particular, the inhibition of voltage-gated Na⁺ currents by AVLE may in part account for the psychopharmacological effects of this herbal remedy.     

Vascular relaxation by ethanol extract of Xanthoceras sorbifolia via Akt- and SOCE-eNOS-cGMP pathways

Aim of the study

The aim of the present study was to define the effect of Xanthoceras sorbifolia extracts (XS) on vascular tension and responsible mechanisms in rat thoracic aortic rings.

Materials and methods

Ethanol extract of the leaves of XS (EXS) was examined for their vascular relaxant effects in isolated phenylephrine-precontracted rat thoracic aorta.

Results

EXS (0.1-100 μg/ml) induced relaxation of the phenylephrine-precontracted aortic rings in a concentration-dependent manner. Endothelium-denudation abolished EXS-induced vasorelaxation. Pretreatment of the endothelium-intact aortic rings with N^G-nitro-l-arginine methylester (l-NAME) and 1H-[1,2,4]-oxadiazolo-[4,3-α]-quinoxalin-1-one (ODQ) inhibited EXS-induced vasorelaxation. Inhibition of Ca²⁺ entry via l-type Ca²⁺ channels failed to block the EXS-induced vasorelaxation. Extracellular Ca²⁺ depletion significantly attenuated EXS-induced vasorelaxation. Modulators of the store-operated Ca²⁺ entry (SOCE), thapsigargin, 2-aminoethyl diphenylborinate (2-APB) and Gd³⁺, and an inhibitor of Akt, wortmannin, markedly attenuated the EXS-induced vasorelaxation. EXS increased cGMP levels of the aortic rings in a concentration-dependent manner and the effect was blocked by l-NAME, ODQ, thapsigargin, Gd³⁺, 2-APB, and wortmannin. Further, EXS-induced vasorelaxation was significantly attenuated by tetraethylammonium, a non-selective K_ca channels blocker, but not by glibenclamide, an ATP-sensitive K⁺ channels inhibitor. Inhibition of cyclooxygenase with indomethacin, and adrenergic and muscarinic receptors blockade had no effects on EXS-induced vasorelaxation.

Conclusions

The present study suggests that EXS relaxes vascular smooth muscle via endothelium-dependent NO-cGMP signaling through activation of the Akt- and SOCE-eNOS-sGC pathways, which may, at least in part, be related to the function of K⁺ channels.     

Protective effects of methanolic extract of Hedyotis puberula (G. Don) R. Br. ex Arn. against experimentally induced gastric ulcers in rat

Aim of the study

This study was aimed to evaluate the antiulcer activity of the whole plants of Hedyotis puberula (G. Don) R. Br. ex Arn.

Materials and methods

Gastroprotective potential of the Hedyotis puberula methanol extract (200 and 400 mg/kg body weight) was studied on indomethacin (IND), ethanol and pyloric ligation (PL)-induced gastric ulcer models in rats.

Results

The treatment with Hedyotis puberula extract at 400 mg/kg p.o. protected the rats against the ulceration which was comparable to the reference drug omeprazole. Pretreatment with extract protected rats from gastric lesion development by way of increased pH and decreased volume, acidity and pepsin content of gastric secretion. Furthermore, total carbohydrate: protein ratio of the gastric juice were noticeably increased in pretreated rats.

Conclusion

Results of our study showed that Hedyotis puberula possess significant gastroprotective activity and validate the folklore claim.     

Hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in carbon tetrachloride induced hepatotoxicity in rats

Ethnopharmacological relevance

In the folk-traditional medicine, snails were used to purify blood, boost immune system, prevent conjunctivitis and to treat liver problems.

Objectives

To evaluate the hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in male Wistar rats treated with carbon tetrachloride as an hepatotoxicant.

Materials and methods

Live adult Bellamia bengalensis was collected commercially from the Kolkata market. Aqueous flesh extract (BBE) was prepared in 0.9% saline and expressed in terms of wet weight basis. The aqueous flesh extract was administered orally (1, 2 g kg⁻¹ day⁻¹) to male rats for 12 days. Liv52 was used as positive control. 24 h after administration of extract, the rats were given a single oral dose of CCl₄ (1.25 ml kg⁻¹), except vehicle control rats. After 24 h of CCl₄ administration, all the animals were sacrificed to collect the blood and liver tissue.

Results

BBE (1 and 2 g kg⁻¹ day⁻¹, p.o. × 12 days) significantly prevented CCl₄ induced elevation of SGOT, SGPT, γGT, ACP, ALP, bilirubin, LDH and CCl₄ induced decrease in total protein, triglyceride level in male Wistar rats. BBE treated rat liver anti-oxidant parameters (LPO, SOD, GSH, CAT, GPx) were significantly antagonized for the pro-oxidant effect of CCl₄. Histopathological studies also supported the protective effect of BBE.

Conclusion

This study validated the folk and traditional use of snail in liver disorder through CCl₄-induced rat experimental model.     

Gastrointestial and respiratory activities of Acacia leucophloea

Ethnopharmacological relevance

The barks of Acacia leucophloea (Fabaceae) are used in Pakistan traditional medicine as an astringent, a bitter, a thermogenic, a styptic, a preventive of infections, an anthelmintic, a vulnery, a demulcent, an expectorant, an antipyretic, an antidote for snake bites and in the treatment of bronchitis, cough, vomiting, wounds, ulcers, diarrhea, dysentery, internal and external hemorrhages, dental caries, stomatitis, and intermittent fevers and skin diseases.

Materials and methods

A study was carried out for the possible elucidation of mechanisms justifying the traditional medicinal uses of A. leucophloea (Fabaceae) in gastrointestinal and respiratory diseases. In vitro experiments were carried out over isolated rabbit jejunum and guinea-pig ileum in order to determine spasmolytic and bronchorelaxant activities, while in vivo studies were conducted in mice for antidiarrheal properties.

Results

A methanol crude extract of barks of the plant caused a concentration-dependent relaxation (0.1-3 mg/ml) of isolated rabbit jejunum preparations in a pattern similar to that of nifedipine and dicyclomine, suggesting a Ca²⁺ channel-blocking mechanism in addition to an anticholinergic effect. In guinea-pig ileum the extract caused a parallel shift in the Ach-curves without suppression of maximum contractile response, followed by a non-parallel shift with the suppression of maximum contractile response at higher concentration similar to that caused by dicyclomine. Moreover, in rabbit trachea, it also caused the relaxation of carbachol (1 μM) and high K⁺-induced contractions at a dose ranging between 0.1578 and 0.734 mg/ml and 0.46-0.94 mg/ml, respectively. These findings indicate that the extract possesses spasmolytic and bronchodilator activities, mediated possibly through blockade of Ca²⁺ channels, thus justifying its medicinal use in diarrhea and asthma. Acacia leucophloea methanol extract exhibited dose-dependent (100-500 mg/ml) protective effect against castor oil induced diarrhea.

Conclusions

The data obtained contribute to the validation of the traditional use of Acacia leucophloea bark in treating gastrointestinal and respiratory disorders, providing an hypothesis on the possible mechanisms of action.     

Cordyceps militaris extract stimulates Cl(-) secretion across human bronchial epithelia by both Ca(2+)(-) and cAMP-dependent pathways

Ethnopharmacological relevance

The caterpillar fungus Cordyceps militaris (CM; Clavicipitaceae) is a well-known traditional Chinese medicine that can be artificially cultivated on a large scale. We have previously demonstrated that its stimulatory action on ion transport in human airway epithelia is similar to Cordyceps sinensis (Clavicipitaceae), which has been traditionally used to treat respiratory diseases.

Aim of the study

To investigate the signal transduction mechanism(s) underlying CM-induced ion transport activity in cultured human bronchial epithelia.

Materials and methods

16HBE14o-, a human bronchial epithelial cell line, was used to study the regulation of ion transport by the water extract of CM. CM extract was added to the apical or basolateral aspect of the epithelia. In subsequent experiments, different Cl⁻ channel and K⁺ channel blockers, adenylate cyclase and protein kinase A (PKA) inhibitors, and an intracellular Ca²⁺ chelator were used to examine the involvement of apical Cl⁻ and basolateral K⁺ channels in mediating CM-induced Cl⁻ secretion and the underlying signal transduction mechanism(s). PKA activity was also measured in 16HBE14o- cells.

Results

CM stimulated Cl⁻ secretion across 16HBE14o- monolayers in a dose-dependent manner. Cl⁻ secretion could be inhibited by apical application of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl⁻channel blocker and the calcium-activated Cl⁻ channel (CaCC) blocker. Cl⁻ secretion was sensitive to basolateral application of different K⁺ channel blockers. Similar inhibitory patterns were obtained in nystatin-permeabilized epithelia. The CM-induced Cl⁻ secretion could be inhibited by adenylate cyclase and PKA inhibitors as well as an intracellular Ca²⁺ chelator. Data from the PKA assay suggested that CM extract caused a significant increase in PKA activity compared with untreated control epithelia.

Conclusions

These results suggest that CM extract stimulated Cl⁻ secretion across human bronchial epithelia, possibly via apical CFTR and CaCC, and the basolateral K⁺ channels are involved in driving apical Cl⁻ exit. The underlying signal transduction mechanisms involve both cAMP- and Ca²⁺-dependent pathways.     

Diuretic activity and acute oral toxicity of Palicourea coriacea (Cham.) K Schum

Aim of the study

Palicourea coriacea (Cham.) K Schum, is an endemic plant used in the Midwestern Region of Brazil, popularly known as “douradinha do campo” and “congonha do campo”. This plant has been used in traditional medicine for several ailments, especially to treat kidney diseases. Since no formal studies on the biological activities and medicinal properties of the ethanolic extract of Palicourea coriacea (PCEE) have been carried out previously, the present study represents the first research into the efficacy of this plant as a diuretic agent employing laboratory rats as test animals.

Materials and methods

For diuretic activity evaluation we assayed three doses of PCEE (20, 40 and 80 mg/kg) and measurement of the urinary volume and electrolytes (Na⁺, K⁺) concentration were taken. The acute oral toxicity of PCEE was investigated according to OECD Guideline 423.

Results

The oral administration of a single dose of PCEE significantly increased the urinary volume in 24 h. Additionally, the treatment with PCEE increased, in a dose-dependent manner, the excretion of both, Na⁺ and K⁺. No sign of toxicity was observed in the animals.

Conclusions

The present study confirmed the ethnopharmacological use of Palicourea coriacea as a diuretic agent in the experimental condition tested here. Additionally, this work supports the importance of the preservation of local knowledge as well as the conservation of Brazilian biodiversity.     

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.     

The antidiarrheal and spasmolytic activities of Phyllanthus emblica are mediated through dual blockade of muscarinic receptors and Ca2+ channels

Aim of the study

This study was aimed at providing the possible mechanisms for the medicinal use of Phyllanthus emblica in diarrhea.

Materials and methods

The in vivo studies were conducted in mice, while isolated rabbit jejunum and guinea-pig ileum were used for the in vitro experiments.

Results

The crude extract of Phyllanthus emblica (Pe.Cr), which tested positive for alkaloids, tannins, terpenes, flavonoids, sterols and coumarins, caused inhibition of castor oil-induced diarrhea and intestinal fluid accumulation in mice at 500-700 mg/kg. In isolated rabbit jejunum, Pe.Cr relaxed carbachol (CCh) and K⁺ (80 mM)-induced contractions, in a pattern similar to that of dicyclomine. The preincubation of guinea pig-ileum with Pe.Cr (0.3 mg/mL), caused a rightward parallel shift in the concentration-response curves (CRCs) of acetylcholine without suppression of the maximum response. While at the next higher concentration (1 mg/mL), it produced a non-parallel rightward shift with suppression of the maximum response, similar to that of dicyclomine, suggesting anticholinergic and Ca²⁺ channel blocking (CCB)-like antispasmodic effect. The CCB-like activity was further confirmed when pretreatment of the tissue with Pe.Cr, shifted the CRCs of Ca²⁺ to the right with suppression of the maximum response, similar to nifedipine or dicyclomine. The activity-directed fractions of Pe.Cr showed a combination of Ca²⁺ antagonist and anticholinergic like components in all fractions but with varying potency.

Conclusion

These results indicate that the Phyllanthus emblica fruit extract possesses antidiarrheal and spasmolytic activities, mediated possibly through dual blockade of muscarinic receptors and Ca²⁺ channels, thus explaining its medicinal use in diarrhea.     

Prevention of experimental diabetes by Uncaria tomentosa extract: Th2 polarization, regulatory T cell preservation or both?

Ethnopharmacological relevance

Uncaria tomentosa (Willd.) DC (Rubiaceae) is a species native to the Amazon rainforest and surrounding tropical areas that is endowed with immunomodulatory properties and widely used around the world. In this study we investigated the immunomodulatory potential of Uncaria tomentosa (UT) aqueous-ethanol extract on the progression of immune-mediated diabetes.

Materials and methods

C57BL/6 male mice were injected with MLDS (40 mg/kg) and orally treated with UT at 10-400 mg/kg during 21 days. Control groups received MLDS alone or the respective dilution vehicle. Pancreatic mononuclear infiltrate and β-cell insulin content were analyzed by HE and immunohistochemical staining, respectively, and measured by digital morphometry. Lymphocyte immunophenotyping and cytokine production were determined by flow cytometry analysis.

Results

Treating the animals with 50-400 mg/kg of UT caused a significant reduction in the glycemic levels, as well as in the incidence of diabetes. The morphometric analysis of insulitis revealed a clear protective effect. Animals treated with UT at 400 mg/kg presented a higher number of intact islets and a significant inhibition of destructive insulitis. Furthermore, a significant protection against the loss of insulin-secreting presented β-cells was achieved, as observed by a careful immunohistochemical evaluation. The phenotypic analysis indicated that the groups treated with higher doses (100-400 mg/kg) presented CD4⁺ and CD8⁺ T-cell values similar to those observed in healthy animals. These same higher doses also increased the number of CD4⁺CD25⁺Foxp3⁺ regulatory T-cells. Moreover, the extract modulated the production of Th1 and Th2, with increased levels of IL-4 and IL-5.

Conclusions

The extract was effective to prevent the progression of immune-mediated diabetes by distinct pathways.     

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.     

Mechanisms of vasorelaxation induced by Ziziphora clinopodioides Lam. (Lamiaceae) extract in rat thoracic aorta

Aim of the study

Ziziphora clinopodioides Lam. (ZC) is widely used in Uyghur folk medicine for the treatment of hypertension diseases in Xinjiang, an autonomous region of China. To provide pharmacological basis for this traditional use, we explored the vasodilating effects of ZC and investigated the underlying mechanisms.

Materials and methods

Activity of hexane (ZCHE), dichloromethane (ZCDE) and aqueous (ZCAE) extracts of ZC were evaluated on isolated rat aortic rings pre-contracted with phenylephrine (PE) or high KCl. The mechanisms were evaluated on ZCDE, the most potent extract.

Results

ZCDE-induced relaxation in endothelium-intact aortic rings pre-contracted with phenylephrine (PE, 10⁻⁶ M) or high KCl (6 × 10⁻² M), with respective EC₅₀ values of 0.27 ± 0.03 and 0.34 ± 0.04 g/l. Mechanic removal of the endothelium did not significantly modify ZCDE-induced relaxation. In endothelium-denuded aorta pre-contracted with PE (10⁻⁶ M), the vasorelaxant effect of ZCDE was significantly decreased by 4-amino-pyridine (10⁻³ M), but not by glibenclamide (10⁻⁴ M), iberiotoxin (3 × 10⁻⁸ M) and thapsigargin (10⁻⁷ M). In Ca²⁺ free solution, ZCDE significantly inhibited extracellular Ca²⁺-induced contraction in high KCl and PE pre-contracted rings. Additionally ZDCE inhibited the intracellular Ca²⁺ release sensitive to PE (10⁻⁶ M).

Conclusions

The results demonstrate that ZDCE exhibits endothelium-independent vasodilating properties that are mediated by inhibition of extracellular Ca²⁺ influx through voltage- and receptor-operated Ca²⁺ channels (VDDCs and ROCCs), by inhibition of Ca²⁺ release from intracellular stores, and also by the opening of voltage-dependent K⁺ channels.     

A modified formulation of Chinese traditional medicine improves memory impairment and reduces Aβ level in the Tg-APPswe/PS1dE9 mouse model of Alzheimer's disease

Ethnopharmacological relevance

SuHeXiang Wan (SHXW), a Chinese traditional medicine has been used orally for the treatment of seizures, infantile convulsion, stroke and so forth. Previously, we reported the effects of modified SHXW essential oil mixture of the fragrance containing herbs on the sedative effect, anticonvulsant property and antioxidative activity after fragrance inhalation.

Materials and Methods

This study was undertaken to evaluate beneficial effects of a modified recipe of SHXW (termed as KSOP1009) consisting of a ethanol extract of 8 herbs including resin of Liquidambar orientalis Miller, seed of Myristica fragrans Houtt., rhizome of Cnidium officinale Makino, lumber of Santalum album L., fructus of Piper longum L., flower buds of Eugenia caryophyllata Merrill et Perry, pollen of Typha orientalis Presl., and root of Salvia miltiorrhiza Bunge in the neurodegenerative diseases such as Alzheimer's disease (AD). The transgenic mice of AD, Tg-APPswe/PS1dE9, were fed KSOP1009 or as a positive control, donepezil for 3 months from 4.5 months of age. Behavioral, immunological and ELISA analyses were used to assess memory impairment, Aβ accumulation and plaque deposition in the brain. Other in vitro works were performed to examine whether KSOP1009 inhibits the Aβ_1-42-induced neurotoxicity in human neuroblastoma cell line, SH-SY5Y cells.

Results

Intake of KSOP1009 improved the Aβ-induced memory impairment and suppressed Aβ levels and plaque deposition in the brain of Tg-APPswe/PS1dE9 mice as much as that of donepezil treatment. KSOP1009 prevented the down-regulation of phospho-CREB and increased AKT phosphorylation in the AD-like brains. Moreover, KSOP1009 suppresses Aβ-induced apoptosis and ROS production in SH-SY5Y cells.

Conclusion

The present study suggests that KSOP1009 may develop as a therapeutic drug for treatment of AD patients.