The neuroprotective effects of an extract of Gastrodia elata

Ethnopharmacological relevance

Gastrodia elata (GE) Blume (family Orchidaceae) is a traditional Chinese herbal medicine for treating headaches, dizziness, tetanus, and epilepsy, indicating neuronal protective functions.

Aim of the study

To evaluate the neuroprotection of GE and its molecular mechanism in preventing serum deprivation-induced PC12 cell apoptosis.

Materials and methods

An MTT assay and Hoechst staining were used to respectively validate serum deprivation-induced cell death and apoptosis. Cyclic (c)AMP formation and protein kinase (PK)A activity were also measured after GE treatment. Western blotting was used to detect the phosphorylation of the cAMP response element-binding (CREB) protein. Transient transfection of a dominant negative CREB was used to validate the importance of CREB.

Results

GE targeted the adenosine A_2A receptor (A_2A-R). GE increased cAMP formation, PKA activity, and phosphorylation of the CREB protein. GE-induced CREB protein phosphorylation and protection was blocked by a PKA inhibitor and overexpression of the dominant negative CREB, respectively.

Conclusions

These results support the neuroprotective effects of GE. The protective mechanism might be mediated through an A_2A-R/cAMP/PKA/CREB-dependent pathway.     

Traditional phytotherapy remedies used in Southern Rwanda for the treatment of liver diseases

Ethnopharmacological relevance

Liver diseases represent a major health problem due to their complications and limited treatment possibilities. In Rwanda, given low accessibility to modern treatments, most people still rely on traditional medicinal plants. The symptomatology of many hepatic troubles (icterus) is evident for traditional healers who have a high probability of selecting efficient herbal medicines.

Objectives

To document medicines used in the treatment of “hepatitis” in Southern Rwanda with the knowledge, attitudes and practices related to liver disorder recognition, control and treatment.

Materials and methods

56 traditional health practitioners, each a legal representative of an official association, were interviewed and participated in plant collection for the preparation of botanically identified herbarium specimens.

Results

68 multi-component and 65 single-component herbal recipes were identified for the treatment of liver diseases with a total of 86 different herbs from 34 families identified. The most represented were the Asteraceae and the Lamiaceae. Crassocephalum vitellinum, Hypoestes triflora and Erythrina abyssinica were the most widely used plants. The principle of polymedication for complex (i.e. multifactorial) diseases (“Ifumbi” in Rwanda), is a constant in every traditional practice. It is striking that the Rwandese therapy of liver diseases proposes so many single-herb preparations (49% of all herbal preparations). Some of the recorded plants or other species from the same genus have previously been documented for liver protection using various in vivo and in vitro models.

Conclusion

Herbal remedies for hepatitis are widely used and highly diverse in Southern Rwanda; further chemical, pharmacological and toxicological studies are clearly required to rationally develop the most important remedies.     

Comparative proteomics research on rat MSCs differentiation induced by Shuanglong Formula

Ethnopharmacological relevance

Shuanglong Formula (SLF) is a classic formula for treating heart disease in traditional Chinese medicine (TCM). And it has been reported that the combinational treatment of SLF with autologous mesenchymal stem cells (MSCs) transplantation could be of benefit to people with acute myocardial infarction. However, their underlying mechanisms are not clear.

Aim of the study

The effects of whole formula and its herbal main ingredients on rat MSCs differentiation towards cardiomyocytes were investigated and the distinct protein expression profile in MSC-derived cardiomyocytes studied.

Materials and methods

The protein expression profiles of rat MSCs and cardiomyocyte-like cells were determined with two-dimensional gel electrophoresis (2-DE).

Results

Our data demonstrated that SLF can induce MSCs into cardiomyocyte-like cells and around 36 proteins mainly involved in cytoskeleton, cell tissue energy metabolism and signal transduction have been shown to be regulated distinctly by SLF treatment.

Conclusions

Data presented in this study suggest that large rearrangement of the proteome occur during the differentiation process of the MSCs to terminally differentiated cardiomyocyte-like cells and offer the possibility for further characterization of specific targets driving the stem cell differentiation.     

Inhibitory effects of Chelidonium majus extract on atopic dermatitis-like skin lesions in NC/Nga mice

Aim of the study

Chelidonium majus (CM) has traditionally been used for treatment of various inflammatory diseases including atopic dermatitis (AD). However its action on atopic dermatitis (AD) is unclear. Therefore, we investigated the effect of CM on AD using NC/Nga mice as an AD model.

Materials and methods

The effect of CM on 1-chloro-2,4-dinitrobenzene (DNCB) induced NC/Nga mice was evaluated by examining skin symptom severity, itching behavior, ear thickness, levels of serum immunoglobulin E (IgE), tumor necrosis factor-α (TNF-α), and interlukin-4 (IL-4), skin histology.

Results

The CM significantly reduced the total clinical severity score, itching behavior, ear thickness and the level of serum IgE in AD mouse model. CM not only decreased TNF-α but also IL-4.

Conclusion

These results suggest that CM may be a potential therapeutic modality for AD.     

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.     

Partially purified extract and viscolin from Viscum coloratum attenuate airway inflammation and eosinophil infiltration in ovalbumin-sensitized mice

Aim of the study

Viscum coloratum Nakai is used in traditional Chinese medicine to treat various diseases, including hemorrhage, hypertension, and inflammatory diseases. A previous study demonstrated a partially purified extract (PPE-SVC) and viscolin from Viscum coloratum Nakai inhibited phosphodiesterase activity. In this study, we evaluated the anti-asthmatic effects of PPE-SVC and viscolin, from Viscum coloratum Nakai, in OVA-sensitized mice.

Materials and methods

Female BALB/c mice were sensitized and challenged with ovalbumin (OVA). The mice were randomized into groups and treated with PPE-SVC, viscolin, or rolipram by intraperitoneal injection on 1 h before each inhalation of OVA and airway hyperresponsiveness (AHR).

Results

PPE-SVC and viscolin suppressed AHR and reduced eosinophil infiltration of the lungs in OVA-sensitized mice. Moreover, PPE-SVC and viscolin inhibited chemokines, including CCL11 and CCL24, and Th2-associated cytokines in bronchoalveolar lavage fluid. However, PPE-SVC and viscolin could not decrease IL-4, IL-5, and IL-13 levels in cultures of OVA-activated spleen cells.

Conclusion

PPE-SVC and viscolin attenuate airway inflammation and eosinophil infiltration in OVA-sensitized mice.     

Ethnopharmacological survey of medicinal herbs in Jordan, the Northern Badia region

Aim of the study

This study sought to gather information from aboriginal Bedouins in North Badia region of Jordan about used medicinal herbs besides their folk uses.

Materials and methods

The data were collected from 40 practitioners who utilized medicinal plants and who were regarded as professional. Subsequently, the uses were compared with the reported ones in the literature. The informant consensus factor (Fic) and use value (UV) have been calculated to those herbs and the managed illnesses.

Results

The data of 73 species were collected; the vast majority of them are safe such as Achillea falcata, Tamarix aphylla and Teucrium polium. Treatment of inflammation and pain presented the major targeted use of these herbs. While, the herbs used for delivery and female problems were limited. This might be due to the culture conservations about the talk of feminine issues. Diseases of kidney, gastrointestinal and respiratory systems as well as diabetes depicted the largest Fic values. Artemisia herba-alba possessed the highest UV value among the studied herbs.     

Andrographolide and 14-deoxy-11,12-didehydroandrographolide from Andrographis paniculata attenuate high glucose-induced fibrosis and apoptosis in murine renal mesangeal cell lines

Ethnopharmacological relevance

Extracts of Andrographis paniculata Nees are used for various ethnomedical conditions including hyperglycemia and hypertension complications.

Aim of the study

The purpose of this study is to evaluate the anti-diabetic nephropathy effect of diterpene lactones andrographolide (AP1) and 14-deoxy-11,12-didehydroandrographolide (AP2) from Andrographis paniculata.

Materials and methods

MES-13, a SV40-transformed murine glomerular mesangial cell line, was cultured in high concentration of glucose to induce diabetic nephropathy phenotypes, which include secretion of extracellular matrix protein fibronectin, cytokine TGF-β, states of oxidative stress, and apoptosis marker caspase-3.

Results

Our data suggest that addition of compounds AP1 or AP2 reduces the phenotypes indicating diabetic nephropathy in MES-13 cells. The compound AP2 showed potent activity than AP1 in the reduction of apoptosis marker caspase-3, fibrosis marker TGF-β, and PAI-1. Furthermore, AP1 and AP2 do not have antioxidant ability in acellular environment; however, addition of AP1 and AP2 reduced intracellular oxidative states in high glucose cultured MES-13 cells.

Conclusion

This is the first report on anti-diabetic nephropathy effect of AP1 and AP2 in part due to the regulation of intracellular signaling transduction, not mere clearance of reactive oxygen species. Thus, this study may be useful for drug development or food supplement for diabetes and nephropathy from Andrographis paniculata.     

Protective effect of hexane extracts of Uncaria sinensis against photothrombotic ischemic injury in mice

Ethnopharmacological relevance

Uncaria sinensis (US) has been used in traditional Korean medicine to treat vascular disease and to relieve various neurological symptoms.

Aim of the study

Scientific evidence related to the effectiveness or action mechanism of US on cerebrovascular disease has not been examined experimentally. Here, we investigated the cerebrovascular protective effect of US extracts on photothrombotic ischemic injury in mice.

Materials and methods

US hexane extracts (HEUS), ethyl acetate extracts (EAEUS) and methanol extracts (MEUS) were administered intraperitoneally 30 min before ischemic insults. Focal cerebral ischemia was induced in C57BL/6J mice and endothelial nitric oxide synthase knockout (eNOS KO) mice by photothrombotic cortical occlusion. We evaluated the infarct volume, neurological score and the activation of Akt and eNOS in ischemic brain.

Results

HEUS more significantly reduced infarct volume and edema than did EAEUS and MEUS following photothrombotic cortical occlusion. HEUS produced decreased infarct volume and edema size, and improved neurological function in a concentration-dependent manner (10, 50, and 100 mg/kg). However, HEUS did not reduce brain infarction in eNOS KO mice, suggesting that the protective effect of HEUS is primarily endothelium-dependent. Furthermore, HEUS (10-300 μg/ml) produced a concentration-dependent relaxation in mouse aorta and rat basilar artery, which was not seen in eNOS KO mouse aorta, suggesting that HEUS cause vasodilation via an eNOS-dependent mechanism. This correlated with increased phosphorylation of Akt and eNOS in the brains of HEUS-treated mice.

Conclusion

HEUS prevent cerebral ischemic damage by regulating Akt/eNOS signaling. US, herbal medicine, may be the basis of a novel strategy for the therapy of stroke.     

Anti-Parkinsonian drug discovery from herbal medicines: what have we got from neurotoxic models?

Ethnopharmacological relevance

Herbal medicines are used to treat Parkinson's disease (PD) in ancient medical systems in Asian countries such as India, China, Japan and Korea based on their own anecdotal or experience-based theories.

Aim of the review

To systematically summarize and analyze the anti-Parkinsonian activities of herbal preparations (including active compounds, herbal extracts and formulations) investigated in the neurotoxic models of PD and provide future references for basic and clinical investigations.

Materials and methods

All the herbal materials tested on in vitro and in vivo neurotoxic models of PD were retrieved from PubMed database by using pre-set searching strings. The relevant compounds and herbal extracts with anti-Parkinsonian activities were included and analyzed according to their chemical classifications or biological activities.

Results

A total of 51 herbal medicines were analyzed. A diversity of compounds isolated from herbal materials were reported to be effective on neurotoxic models of PD by modulating multiple key events or signaling pathways implicated in the pathogenesis of PD. The main structure types of these compounds belong to catechols, stilbenoids, flavonoids, phenylpropanoids and lignans, phenylethanoid glycosides and terpenes. Although some herbal extracts and formulations have shown positive results on PD animal models, the relative compounds accounting for the effects and the underlying mechanisms remain to be further investigated.

Conclusions

Herbal medicines can be an alternative and valuable source for anti-Parkinsonian drug discovery. Compounds classified into stilbenoids, flavonoids, catechols and terpenes may be the most promising candidates for further investigation. Some well-studies compounds such as baicalein, puerarin, resveratrol, curcumin and ginsenosides deserve further consideration in clinical trials. In-depth experimental studies are still needed to evaluate the efficacy of herbal extracts and formulations in PD models.     

Anti-amnesic effect of Chong-Myung-Tang on scopolamine-induced memory impairments in mice

Aim of the study

Chong-Myung-Tang (CMT) consisted of Acorus gramineus Soland, Polygala tenuifolia Willdenow, and Poria cocos Wolf is one of the traditional Korean herbal medicines used for the therapy of learning and memory improvement. The present study was investigated the effect of CMT on learning and memory functions in SCOP-induced memory deficits mice.

Materials and methods

The cognitive-enhancing effect of CMT on amnesic mice induced by SCOP was investigated by assessing the passive avoidance test and the Morris water maze test. In order to confirm the underlying mechanisms of memory enhancing effects of CMT, activities of AChE, choline acetyltransferase (ChAT), and antioxidant enzymes were measured.

Results

Administration of CMT significantly restored memory impairments induced by SCOP in the passive avoidance test and also reduced escape latency during trial sessions in the Morris water maze test. The increased AChE activity produced by SCOP was significantly inhibited by CMT. CMT significantly enhanced ChAT activity. Moreover, treatment with CMT to the amnesic mice induced by SCOP considerably decreased malondialdehyde levels and restored activities of superoxide dismutase and catalase to the control values.

Conclusions

These results suggest that CMT may be useful for the cognitive improvement via regulation of cholinergic marker enzyme activities and the antioxidant defense system.     

Evaluation of the acute dermal exposure of the ethanolic and hexanic extracts from leaves of Schinus molle var. areira L. in rats

Ethnopharmacological relevance

Schinus molle var. areira L. (Anacardiaceae) is employed in herbal medicine for many conditions, including respiratory, urinary and menstrual disorders, and as a digestive stimulant, diuretic, astringent and antidepressant. It is also known for its topical use as wound healer, antiseptic, for skin disorders and as repellent and insecticide. In the present work, the acute dermal exposure to ethanolic and hexanic extracts from leaves of Schinus molle var. areira was studied in rats.

Materials and methods

A single dose of 2000 mg/kg of body weight of ethanolic and hexanic extracts from leaves was applied on the shaved skin of male and female rats. After 24 h of exposure, the patch was removed and any sign of irritation was recorded. Behavioral and functional parameters in a functional observational battery and motor activity in an open field were assessed after the exposure to the extracts. Then, after 14 days of observation, animals were retested. Finally, histopathological studies were conducted on several organs.

Results

Slight signs of erythema and edema were observed in the skin site of exposure, but they disappeared after 48 h. The exposure to the hexanic extract produced an increase in parameters of activity, rearing and arousal assessed in the functional observational battery, which reversed after 14 days. On the other hand, the ethanolic extract caused an increase in locomotor activity, reflected in a higher number of rearings performed in the open field in the evaluation carried out on Day 14. No histopathological alterations were detected in the analyzed organs.

Conclusions

The results show that the acute dermal exposure of the ethanolic and hexanic extracts from leaves of Schinus molle var. areira only causes a slight and reversible skin irritation, and a mild stimulatory effect in rats. All these indicate that the topical use of these extracts would be safe.     

The herbal market of Cyprus: traditional links and cultural exchanges

Aim of the study

Directive 2004/24/EC sets the frame for the herbal trade in Europe emphasizing the need of the botanical characterization of herbal substances intended for human consumption. Thus, the aim of the present study is to survey and identify taxonomically the herbs sold in the market of Cyprus and to document and evaluate their uses. Moreover, the relationship of the nowadays market to the local ethnobotanical tradition, as well as to other eastern Mediterranean markets (i.e. Greece, Israel, Jordan, Turkey) is also traced.

Material and methods

Herbs sold in 15 shops scattered in three major cities of Cyprus were surveyed. Data concerning their uses were collected through interviews with the herbal traders and studied quantitatively by the use of ethnobotanical indices (informant consensus factor, use value, relative importance value, fidelity level) and statistical tests (Spearman's correlation coefficient, Kruskal-Wallis test, paired-sample t-test). Comparisons to published information concerning other eastern Mediterranean markets were performed by multivariate tests (Ward's hierarchical cluster).

Results

A total number of 57 taxa were surveyed from 15 selected shops, out of which 32 are cultivated, 14 are collected from the wild and 11 are imported. The highest Informant Consensus Factors were presented by the neuropsychiatric (0.78), gastrointestinal (0.76) and respiratory ailments (0.71). The comparison of our results to published information has shown that 40 of the traded taxa have also been used traditionally, while a considerable number of herbs are also found in the markets of Greece and Turkey (35 and 29 taxa, respectively); a lower number of the taxa traded, is found in the markets of the Middle East.

Conclusions

The nowadays herbal trade in Cyprus is still linked to the local ethnobotanical legacy. Moreover the similarities between the Cypriot herbal market and the markets of Greece and Turkey are more prominent than those between Cyprus and the Middle East, reflecting the cultural exchanges among the three neighbouring countries.     

Involvement of heme oxygenase-1 in the anti-inflammatory activity of Chrysanthemum boreale Makino extracts on the expression of inducible nitric oxide synthase in RAW264.7 macrophages

Aim of the study

This study is to elucidate the involvement of anti-inflammatory heme oxygenase-1 (HO-1) in the inhibitory activity of a Chrysanthemum boreale Makino (CB) extract on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages.

Materials and methods

Cell viability and NO assay were performed. In addition, iNOS expression was detected by Western blotting and real-time PCR. HO-1 expression was also evaluated by Western blotting, and blocking HO-1 activity on NO production was performed.

Results

The CB extract at the highest concentration (100 μg/ml) significantly inhibited NO production by approximately 90% and suppressed iNOS protein expression by approximately 84.8% compared to LPS-stimulated cells. Furthermore, the CB extract (100 μg/ml) inhibited iNOS mRNA expression in a concentration-dependant manner and suppressed iNOS mRNA expression by 94.8%. The CB extract induced the expression of HO-1 in a dose-dependent manner, and blocking HO-1 activity abolished the inhibitory effects of the CB extract. Moreover, the addition of carbon monoxide such as tricarbonyl dichlororuthenium (II) dimmer (RuCO), a byproduct derived from heme degradation, mimicked the inhibitory action of low concentrations of CB extract.

Conclusion

These results suggest that a CB extract has potent anti-inflammatory activity in RAW264.7 macrophages involving the induction of HO-1.