Κυριακή 16 Ιουνίου 2019

Ethnopharmacology

Toxicological findings about an anticancer fraction with casearins described by traditional and alternative techniques as support to the Brazilian Unified Health System (SUS)

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Paulo Michel Pinheiro Ferreira, Denise Barbosa Santos, Jurandy do Nascimento Silva, Amanda Freitas Goudinho, Carla Lorena Silva Ramos, Patrícia Canteri de Souza, Ricardo Sérgio Couto de Almeida, Diego Sousa Moura, Rhaul de Oliveira, Cesar Koppe Grisolia, Alberto José Cavalheiro, Ana Amélia de Carvalho Melo-Cavalcante, José Roberto de Oliveira Ferreira, Manoel Odorico de Moraes Filho, Claudia Pessoa

Abstract
Ethnopharmacological relevance

Extracts, essential oils and molecules from Casearia sylvestris have popularly shown pharmacological actions against chronic diseases, as anxiety, inflammation, cancer and dyslipidemia. In the context of antitumoral therapy, we investigated in vitroex vivo and in vivo toxicological changes induced by a Fraction with Casearins (FC) and its component Casearin X isolated from C. sylvestris on animal and vegetal cells, and upon invertebrates and mammals.

Material and methods

Cytotoxicity was carried out using normal lines and absorbance and flow cytometry techniques, Artemia salina nauplii, Danio rerio embryos and meristematic cells from Allium cepa roots. Acute and 30 days-mice analysis were done by behavioral, hematological and histological investigations and DNA/chromosomal damages detected by alkaline Cometa and micronucleus assays.

Results

FC was cytotoxic against lung and fibroblasts cells and caused DNA breaks, loss of integrity and mitochondrial depolarization on ex vivo human leukocytes. It revealed 24 h-LC50 values of 48.8 and 36.7 μg/mL on A. salinanauplii and D. rerio embryos, reduced mitotic index of A. cepa roots, leading to cell cycle arrest at metaphase and anaphase and micronuclei. FC showed i.p. and oral LD50 values of 80.9 and 267.1 mg/kg body weight. Subacute i.p. injections induced loss of weight, swelling of hepatocytes and tubules, tubular and glomerular hemorrhage, microvesicular steatosis, lung inflammatory infiltration, augment of GPT, decrease of albumin, alkaline phosphatase, glucose, erythrocytes, and lymphocytes, and neutrophilia (p > 0.05). FC-treated animals at 10 mg/kg/day i.p. caused micronuclei in bone marrow and DNA strand breaks in peripheral leukocytes.

Conclusions

This research postulated suggestive side effects after use of FC-related drugs, demonstrating FC as antiproliferative and genotoxic on mammal and meristematic cells, including human leukocytes, teratogenicity upon zebrafish embryos, myelosuppression, clastogenicity, and morphological and biochemical markers indicating liver as main target for FC-induced systemic toxicity.

Graphical abstract

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Neoline is the active ingredient of processed aconite root against murine peripheral neuropathic pain model, and its pharmacokinetics in rats

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Yohei Tanimura, Masato Yoshida, Kan'ichiro Ishiuchi, Masahiro Ohsawa, Toshiaki Makino

Abstract
Ethnopharmacological relevance

Processed aconite root (PA), the root of Aconitum carmichaeli (Ranunculaceae), is a crude drug used in traditional Chinese or Japanese kampo medicine to treat pain associated with coldness. In our previous study, PA and its active ingredient, neoline, alleviated oxaliplatin-induced peripheral neuropathy in mice.

Aim of the study

The present study investigated the effects of PA on a murine peripheral neuropathy model induced by intraperitoneal injection of paclitaxel and partial ligation of the sciatic nerve (Seltzer model), and identified its active ingredients.

Materials and methods

PA powder (1 g/kg/day) was orally administered, and either neoline or benzoylmesaconine (10 mg/kg/day) was subcutaneously injected into the murine model. Mechanical hyperalgesia was evaluated via the von Frey filament method. PA extract was orally administered to rats; blood samples were chronologically collected, and the plasma concentrations of Aconitum alkaloids were measured. The contents of Aconitum alkaloids in commercial PA products were also measured.

Results

PA extract and neoline significantly attenuated the mechanical hyperalgesia induced by either paclitaxel or partial ligation of the sciatic nerve in mice. In the plasma samples of rats treated with PA extract, higher concentrations of benzoylmesaconine and neoline were apparent among Aconitum alkaloids. The contents of benzoylmesaconine and neoline varied among PA products with different processing procedures. Subcutaneous injection of benzoylmesaconine did not attenuate the hyperalgesia induced by each paclitaxel, partial ligation of the sciatic nerve, or oxaliplatin in mice.

Conclusions

The present results indicate that PA and its active ingredient, neoline, are promising agents for the alleviation of neuropathic pain. Neoline can be used as a marker compound to determine the quality of the PA products for the treatment of neuropathic pain.

Graphical abstract

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Anti-aggregant effect of butanolic extract of Rubia tinctorum L on platelets in vitro and ex vivo

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Fatima Zahra Marhoume, Mehdi Ait Laaradia, Younes Zaid, Jawad Laadraoui, Sara Oufquir, Rachida Aboufatima, Abderrahmane Chait, Abdallah Bagri

Graphical abstract

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Antibacterial and antidiarrheal activity of Butea Monospermea bark extract against waterborne enterobacter Cloacae in rodents: In-vitro, Ex-vivo and In-Vivo evidences

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Divya Sharma, Saraswati Patel, Kanika Verma, Shiva Gudlawar, Dipjyoti Chakraborty, Sarvesh Paliwal, Jaya Dwivedi, Swapnil Sharma

Abstract
Ethnopharmacological relevance

Butea monosperma (Lam.) Taub. (family Leguminosae), popularly known as 'Palash' possess numerous medicinal properties since ancient times. According to the Wealth of India, stem bark of this plant exhibits various therapeutic properties like antimicrobial, astringent, styptic, aphrodisiac, and anti-inflammatory.

Aim of the study

The purpose of the present study was to investigate antibacterial and antidiarrheal effect of B. monosperma bark against newly isolated gram negative pathogenic bacterial strain Enterobacter cloacae.

Materials and methods

Aqueous extract of B. monosperma bark (BMAqE) was subjected to LC-MS/MS analysis for determination of bioactive components. Antibacterial study of BMAqE was assessed using bacterial growth kinetic study, fluorescence spectroscopy, outer and inner membrane permeability assay, dehydrogenase inhibitory assay and protein leakage assay followed by field emission scanning electron microscope (FE-SEM) study. Antidiarrheal activity was studied using castor oil induced diarrhea model in albino rats followed by histopathology studies of rat ileum.

Results

LC-MS/MS analysis of BMAqE revealed presence of twenty-two different active phytoconstituents out of which most of the constituents belong to flavonoid and polyphenol family. BMAqE showed MIC and MBC (IC90) value of 5 and 200 μg/mL against targeted bacterial strain. BMAqE exhibited potent and dose dependent bactericidal effect via disruption of integrity of bacterial cell membrane, enzymatic degradation, leakage of intracellular protein and ruptured bacterial cell. In castor oil induced diarrhea model, BMAqE (200 mg/kg; orally) caused marked reduction (75.66%) in the frequency of defecation and mean weight of faeces (0.54 ± 0.04) when compared to control group (2.26 ± 0.25). Histopathology study revealed marked restoration of cellular architecture of rat ileum tissue. Four known flavonoids were isolated from BMAqE using column chromatography. In ex-vivo study, BMAqE (0.0002, 0.0004 and 0.0006 g/L) and isolated flavonoids i.e. rhamnetin, quercetin, kaempferol and catechin (0.5, 5 & 50 μm) produced a significant (p < 0.001) change in EC50 and indicated competitive phenomena via rightward shift of acetylcholine CRC with pA2 of 3.78, 8.0, 7.1, 7.0 and 6.9 respectively.

Conclusion

BMAqE exhibits impressive antibacterial and anti-diarrheal activity and can be effectively used to eradicate water borne diseases.

Graphical abstract

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The contribution of beta-2 adrenergic, muscarinic and histamine (H1) receptors, calcium and potassium channels and cyclooxygenase pathway in the relaxant effect of Allium cepa L. on the tracheal smooth muscle

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Arghavan Memarzia, Fatemeh Amin, Saeideh Saadat, Mehrdad Jalali, Zahra Ghasemi, Mohammad Hossein Boskabady

Abstract
Ethnopharmacological relevance

There are report regarding therapeutic effects for Allium cepa L. (A. cepa) in Iranian traditional medicine and the plant has showed anti-inflammatory, anti-allergic, anti-hyperglycemic, antioxidant, anti-cancer, anti-hypertension, anti-hypercholesterolemia and anti-asthmatic activities in previous studies.

Aim of the study

In this study, the contribution of β2 adrenergic, muscarinic and histamine (H1) receptors, calcium and potassium channels, and cyclooxygenase pathway in the relaxant effect of A. cepa extract on tracheal smooth muscle (TSM) was assessed.

Materials and methods

TSM was contracted by KCl (60 mM) or methacholine (10 μM) for 5 min and cumulative concentrations of A. cepa extract (2, 4, 8, 16, 32 and 64 mg/ml) were added to organ bath every 5 min. Theophylline (0.2, 0.4, 0.6 and 0.8 mM) as positive control, and saline (1 ml) as negative control were also examined in non-incubated tissues. The relaxant effect of A. cepa extract was examined on non-incubated and incubated TSM with propranolol, chlorpheniramine, diltiazem, atropine, glibenclamide and indomethacin.

Results

A. cepa showed concentration-dependent relaxant effects on non-incubated TSM contracted by KCl (60 mM) or methacholine (10 μM), (P < 0.01 to p < 0.001). There was no significant difference in the relaxant effects of A. cepa between non-incubated and incubated tissues with glibenclamide, atropine, chlorpheniramine and indomethacin. The plant extract showed significant lower relaxant effects in incubated TSM with propranolol and diltiazem compared to non-incubated tissues. EC50 values of A. cepa extract in incubated TSM with propranolol and diltiazem were significantly lower than non-incubated tissues (p < 0.001 and p < 0.05, respectively). The relaxant effects of different concentrations of the extract of A. cepa were not significantly different with those of theophylline. The concentrations of A. cepa extract and theophylline were significant correlated with their relaxant effects (p < 0.05 to p < 0.001). In incubated TSM with propranolol and diltiazem, concentration ratio minus one (CR-1) values was positive (2.65 ± 0.63 and 1.28 ± 0.43 respectively).

Conclusion

The A. cepa extract showed relatively potent relaxant effect on TSM which was comparable to the effect of theophylline. The results showed that β2-adrenergic stimulatory and/or calcium channel blockade are the possible mechanisms for the relaxant effects of the plant.

Graphical abstract

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Ethnobotanical survey of medicinal plants in central Abyan governorate, Yemen

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Mohamed Al-Fatimi

Abstract
Ethnopharmacological relevance

Traditional medicinal plant knowledge in Yemen is still only passed on orally from one generation to another. A few studies, mostly very limited in scope, have been dedicated to this indigenous knowledge so far. This paper presents the results of the first extensive field study in southern mainland Yemen, undertaken in three communities in the central region of Abyan governorate. The study is aimed to preserve the heritage of this indigenous knowledge and to explore and select local medicinal plants that promise high pharmacological efficacy for further pharmacological and phytochemical investigations.

Methods

356 indigenous informants (273 males, 83 females) were interviewed about the modes of application and uses of the medicinal plants in the region. The ethnobotanical data were recorded by semi-structured face-to-face interviews, substantiated by specimen collections and taxonomic identifications and quantitative data analysis including informant consensus factor (ICF) and number of use-reports.

Results

In total, 195 medicinal plant species (170 wild and 25 cultivated) belonging to 138 genera and 55 flowering plant families, were recorded for the treatment of 155 different ailments classified into 16 categories. Apocynaceae (25 species), Fabaceae (18 species), Euphorbiaceae (16 species) and Asteraceae (14 species) were the most frequently used plant families. Paste (86) followed by unprepared plant exudates (77) and decoction (55) were the most common herbal preparation modes. Dermal application (51.8%) was the most common administration route. The highest number of use reports and the maximum number of used medicinal plants were recorded for treatment of skin and gastrointestinal ailments. Forty-five species have never been reported in the ethnomedicinal literature before. Plant species with the most citations were Aloe vacillans Forssk. (malaria), Solanum incanum L. (tooth decay), Caralluma awdeliana (Deflers) A. Berger (diabetes), Tribulus terrestris L. (kidney stones), Aristolochia bracteolata Lam. (snake poison), Hydnora abyssinica A.Br. (stomach ulcer), Indigofera oblongifolia Forssk. (urine retention) and Chrozophora oblongifolia (Delile) A. Juss ex Spreng (haemorrhoids).

Conclusions

Significant traditional knowledge of the uses of local medicinal plant species was recorded for the first time for central Abyan in particular and southern Yemen in general. The study shows that the medicinal plants still play an important role in the primary health care in the study area. The ethnobotanical results provide a basis for further pharmacological, biological, pharmacognostical, and phytochemical investigations.

Graphical abstract

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A lipid-soluble extract of Pinellia pedatisecta Schott enhances antitumor T cell responses by restoring tumor-associated dendritic cell activation and maturation

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Yumeng Wang, Haixia Huang, Sheng Yao, Guiling Li, Congjian Xu, Yang Ye, Suiqi Gui

Abstract
Ethnopharmacological relevance

Pinellia pedatisecta Schott (PPS)is a traditional Chinese medicine functioning as reducing swelling and drying dampness. Pinellia pedatisecta Schott extract (PE) has been confirmed to suppress cervical tumor growth and modulate the antitumor CD4+T helper immunity towards Th1.

Aims

To explore the roles of PE in modulating tumor-associated dendritic cell (TADC) activation and function.

Methods

For in vivo studies, HPV+TC-1 mouse tumor models were conducted and treated with PE for 3 weeks (10 mg/kg/d or 20 mg/kg/day). The immune profiles of spleen, tumor-draining lymph nodes (TDLNs), tumor and serum were analyzed by flow cytometry and multiplexed bead-based immunoassay. For in vitro studies, TADCs were generated by tumor-conditioned medium and treated with PE solution. The maturation and function of TADCs were evaluated by flow cytometry, ELISA, mixed lymphocyte reaction (MLR) and cytotoxic T lymphocyte (CTL) assay. Furthermore, the effect of PE on SOCS1 pathway was examined by western blotting and real time PCR.

Results

PE upregulated the expression of major histocompatibility complex class II (MHCII) and costimulatory molecules CD80 and CD86 on TADCs and promoted IL-12 secretion from TADCs. In addition, PE-treated TADCs promoted the proliferation of CD4+ and CD8+ T cells and induced the differentiation of IFN-γ+CD4+ and GZMB+CD8+ T cells. PE-treated TADCs also elicited a more powerful antigen-specific cytotoxic T lymphocyte (CTL) response. Furthermore, PE treatment in vivo enhanced the proliferation, activated the functional ability (increased Ki67, CD137, GZMB or IFN-γ, TNF-α expression) and reversed the exhaustion (impaired CD95 or PD-1 expression) of antitumor T cells. Mechanistically, PE inhibited SOCS1-restrained JAK2 activation in TADCs.

Conclusions

PE efficiently restored the immature status of TADCs and enhanced their function as antigen-presenting cells to further elicit antitumor Th1 and CTL responses, suggesting that PE may be a potential immunomodulatory drug for cancer treatment.

Graphical abstract

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Binding of the polysaccharide from Acanthopanax giraldii Harms to toll-like receptor 4 activates macrophages

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Qingqing Li, Zhiting Chen, Zhilu Xu, Shaoyun Han, Huihui Hao, Jiang Wu, Fengxiang Sun, Xiaoyan Fu, Ruyue Li, Birong Zheng, Xiaoxiao Guo, Tongtong Zhang, Yong Chen

Abstract
Ethnopharmacological relevance

The traditional Chinese medicine, Acanthopanax giraldii Harms, is commonly used to treat arthralgia due to wind, cold and dampness, as well as weakness in the feet and knees. Its other reported effects include eliminating flatulence, strengthening muscles and bones, and delaying aging. The polysaccharides in A. giraldii Harms are the major bioactive substances that confer the herb's antioxidant properties as well as anticancer and antiviral effects.

Aims of the study

To elucidate the underlying mechanism and signaling cascade involved in the homogeneous A. giraldii Harms polysaccharide II (AHP–II)–mediated immunomodulation of mice macrophages.

Materials and methods

The phagocytosis of neutral red and the production of nitric oxide, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), were measured to determine AHP–II–induced macrophage activation. Confocal microscopy and flow cytometry were used to confirm the binding of AHP-II to macrophages. The involvement of Toll-like receptor (TLR) 4 in AHP–II–induced macrophage activation was demonstrated using antibody blocking and macrophages from C3H/HeJ TLR4-mutant mice. Western blotting was used to map AHP–II–induced downstream signaling pathways.

Results

AHP-II increased the phagocytosis of macrophages and the release of nitric oxide, IL-6 and TNF-α cytokines. Direct, saturable and reversible binding of AHP-II to macrophages was observed, while it can be inhibited by the anti-TLR4 antibody. In addition, the presence of the anti-TLR4 antibody inhibited AHP–II–induced macrophage IL-6 and TNF-α production in the peritoneal macrophages of C3H/HeJ mice. Moreover, AHP–II–TLR4-stimulated macrophages activate the downstream intracellular ERK and JNK/nuclear factor (NF)-κB signaling pathways. In addition, the AHP–II–mediated regulation of IL-6 and TNF-α production from macrophages was greatly affected by specific ERK, JNK and NF-κB inhibitors.

Conclusion

Our study elucidated the immunomodulatory mechanism of AHP-II in macrophage activation and identified TLR4 as the main receptor coordinating AHP-II binding. Our findings suggest AHP-II may be used as a novel immunopotentiator for medical purposes.

Graphical abstract

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Utilization trends in traditional Chinese medicine for acute myocardial infarction

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Chung-Yen Lu, Pei-Chin Lu, Pei-Chun Chen

Abstract
Ethnopharmacological relevance

Among heart diseases, acute myocardial infarction (AMI) is the most serious and life-threatening emergency. In Taiwan, heart disease has consistently ranked second among the top 10 leading causes of death since 2007, second only to malignant tumors; however, population-based studies on the use of traditional Chinese medicine (TCM) in AMI cases are limited.

Aim of the study: This study investigated the characteristics of TCM users and prescriptions of TCM, and their differences between two cohorts of patients with AMI, identified 10 years apart.

Materials and methods

A cross-sectional study was conducted using the Taiwan National Health Insurance claims database. From among two random sample of 1 million beneficiaries selected from the claims database, we identified two cohorts of patients with first hospitalization for AMI in between 2000–2001 and 2010–2011. Patients who had received TCM therapy within one year after hospital discharge were defined as TCM users, whereas, all the other patients with AMI were considered non-users of TCM. We compared the characteristics of TCM use and the patterns of prescriptions between the two cohorts.

Results

The proportion of patients receiving TCM care was similar between the two AMI cohorts; 20% (85/418) of the patients were diagnosed in 2000–2001 and 21% (169/817) in 2010–2011. In the 2010–2011 AMI cohort, the proportion of men was smaller among TCM users than non-users, and TCM users were less likely to have hyperlipidemia. Among TCM users, the most frequently prescribed herb was Dan-shen (Salvia miltiorrhiza Bunge, Salvia root) in both cohorts. The most commonly used Chinese herbal formulations were Xue-Fu-Zhu-Yu-Tang (Blood Mansion Dispel Stasis) for the 2000–2001 cohort and Zhi-Gan-Cao-Tang (Honey-Fried Licorice Decoction) for the 2010–2011 cohort.

Conclusions

This study revealed the differences in the prescription frequency of Chinese herbal formulation among the two cohort of patients with AMI, suggesting that the practice of prescribing TCM has evolved from post-antique formula to classical remedies during the 10 years evaluated. Further investigations are needed to evaluate if the change in the utilization of Chinese herbal formulations impacts the effectiveness of the treatment.

Graphical abstract

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Rhamnus prinoides (gesho): A source of diverse anti-biofilm activity

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Mariya Campbell, Weilun Zhao, Raghda Fathi, Merhawi Mihreteab, Eric S. Gilbert

Abstract
Ethnopharmacological relevance

Rhamnus prinoides (gesho) is an evergreen shrub from East Africa traditionally used for the treatment of illnesses including atopic dermatitis, ear, nose and throat infections, pneumonia, arthritis, brucellosis, flu, indigestion and fatigue.

Aim of the study

Several of the conditions for which gesho is traditionally used are associated with communities of surface-attached microorganisms, or biofilms. We hypothesized that gesho has anti-biofilm activity. The principal aim of this study was to evaluate gesho-associated anti-biofilm activity and identify active compounds.

Materials and methods

Lyophilized ethanol and aqueous extracts were prepared from dried Rhamnus prinoides stems and leaves. Biofilm inhibition was measured by crystal violet staining and subsequent viability assays were conducted on growth agar. Chemical fractionation, chemical testing, Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS) were used to isolate and identify active compounds.

Results

Leaf and stem ethanol extracts significantly inhibited Staphylococcus aureusBacillus subtilis and Streptococcus mutans biofilm formation up to 99.9% and reduced planktonic cell growth up to 10 log units relative to untreated controls. The anti-biofilm activity of the ethanol stem extracts was due to a biocidal or bacteriostatic mechanism while bacteriostatic or anti-pathogenic mechanisms were attributed to the leaf ethanol extract. Gesho extracts showed activity against all three species tested but the treatment efficacy and mechanism were species dependent. Chemical fractionation and activity screens of the leaf ethanol extract identified ethyl 4-ethoxybenzoate and 4-hydroxy 4-methyl pentanone to be compounds with anti-biofilm activity. Ethyl 4-ethoxybenzoate activity was potentiated by DMSO. Notably, concentrations of both compounds were identified where biofilm formation was prevented without inhibition of cell growth; i.e. anti-pathogenic characteristics were evident.

Conclusion

Gesho leaf ethanol extract contains chemicals with anti-biofilm and bactericidal activities. This work lends support to the traditional use of gesho for treating topical infections and warrants further investigation into Rhamnus prinoides as a source of antibacterial and anti-biofilm agents.

Graphical abstract

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Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
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