Ethanol Extract of Anacyclus pyrethrum Root Ameliorates Cough-Variant Asthma Through the TLR4/NF-κB Pathway and Wnt/ß-Catenin Pathway.

Cough-variant asthma (CVA) has been recognized as the initial stage or pre-asthmatic state of classic asthma, which characterized by cough as the primary clinical presentation. Inhaled glucocorticoids, oral leukotriene receptor antagonists and antihistamines are the clinical treatments, but their efficacy is not satisfactory. Some traditional Chinese medicine (TCM) has been reported to have certain advantages in the treatment of CVA, but the underlying molecular mechanisms are still unclear. Recent research has indicated that Anacyclus pyerhrurm (L) DC. is commonly used in the treatment of human diseases. The aim of our study was to evaluate the anti-inflammatory and anti-oxidative mechanism of the ethanol extract of Anacyclus pyrethrum (L) DC. root (EEAP) in a model of CVA. In our study, we indicated that EEAP ameliorated CVA by reducing cough frequency and inflammatory effect and oxidative stress in an in vivo rat model of CVA. In addition, EEAP ameliorated LPS-induced cell apoptosis and regulated inflammatory effect and oxidative stress in vitro. Mechanistically, EEAP exerted anti-inflammatory effects through regulating the TLR4/NF-κB pathway and Wnt/ß-catenin pathway, and overexpressing TLR4 or activating the Wnt/ß-catenin pathway by SKL2001 reversed EEAP-exerted effects in LPS-exposed BEAS-2B and 16-HBE cells. In conclusion, EEAP attenuated cell apoptosis, inflammation and oxidative stress through restraining the TLR4/NF-κB pathway and Wnt/ß-catenin pathway in CVA, which shown that EEAP might be a promising therapeutic agent for CVA and may provide a theoretical basis for clinical treatment with CVA patients.

Phytochemical, Morphological and Genetic Characterisation of Anacyclus pyrethrum var. depressus (Ball.) Maire and Anacyclus pyrethrum var. pyrethrum (L.) Link.

The present study is based on a multidisciplinary approach carried out for the first time on Anacyclus pyrethrum var. pyrethrum and Anacyclus pyrethrum var. depressus, two varieties from the endemic and endangered medicinal species listed in the IUCN red list, Anacyclus pyrethrum (L.) Link. Therefore, morphological, phytochemical, and genetic characterisations were carried out in the present work. Morphological characterisation was established based on 23 qualitative and quantitative characters describing the vegetative and floral parts. The phytochemical compounds were determined by UHPLC. Genetic characterisation of extracted DNA was subjected to PCR using two sets of universal primers, rbcL a-f/rbcL a-R and rpocL1-2/rpocL1-4, followed by sequencing analysis using the Sanger method. The results revealed a significant difference between the two varieties studied. Furthermore, phytochemical analysis of the studied extracts revealed a quantitative and qualitative variation in the chemical profile, as well as the presence of interesting compounds, including new compounds that have never been reported in A. pyrethrum. The phylogenetic analysis of the DNA sequences indicated a similarity percentage of 91%. Based on the morphological characterisation and congruence with the phytochemical characterisation and molecular data, we can confirm that A. pyrethrum var. pyrethrum and A. pyrethrum var. depressus represent two different taxa.

Phytochemistry, Biological and Pharmacological Activities of the Anacyclus pyrethrum (L.) Lag: A Systematic Review.

Anacyclus pyrethrum (L.) (Asteraceae) is an important annual medicinal herb and is widespread in Morocco and Algeria. Most of its parts are used in traditional medicine and the roots are the most important parts used. The present review gives an account of the updated information on its phytochemical and pharmacological properties. We have collected the essential characteristics and the different scientific data of the A. pyrethrum species, and reviewed its potential. It is seen from the literature that A. pyrethrum is a rich source of the phytochemical constituents such as alkaloids (pellitorin) and n-alkylamides. This species also contains pyrethrins, sesamin, traces of essential oils and a wide range of other chemical compounds. These active substances possess antimicrobial and anti-inflammatory activities. The plant has an antidiabetic, insecticidal and immunostimulatory effect, as well as an aphrodisiac and antioxidant potentials, and various other important medicinal properties. Many traditional uses are also reported in previous research such as for rheumatism, sciatica, colds, neuralgia and paralysis. This species is considered to be a sialagogue, and used in the treatment of stomach ailments, diseases of inflammation of the mouth, against cysts in the genital tract and to relieve toothaches. Thus, further research must be carried out in order to establish any relationship between the traditional uses, phytochemistry and toxicity. Moreover, A. pyrethrum is quite promising as a medicinal agent, so further clinical trials should be performed to prove its efficacy.

The Hepatoprotective and Anti-Nephrotoxic Potential of Methanolic Extract of a Polyherbal Preparation in CCl4-Induced Liver Injury Model of Wistar Rats.

The liver and kidneys are the vital organs of the body and perform important life-sustaining functions in the body. Synthetic drugs used in the treatment of liver and kidney diseases are sometimes inadequate and can lead to serious side effects. Medicinal herbs and plants were used to combat diseases for a long time and combination therapy is preferred over single plant therapy. In the current study, the Asparagus racemosus, Mucuna pruriens, Anacyclus pyrethrum, and Tribulus terrestris polyherbal preparation (PHP) was selected to evaluate its hepatoprotective, antioxidant, and anti-nephrotoxic potential. The methanolic extract of PHP was prepared following standard protocols. Fifty-six albino rats were divided into 7 groups (n = 8). The negative control (NC) having the healthy rats and the remaining 6 groups were induced liver toxicity by intraperitoneally injecting 0.5 mL/kg of 50% CCl4 in olive oil. Group 2 was positive control and group 3 and 4 received silymarin standard drug at the dose of 100 and 200 mg/kg body weight. Groups 5, 6, and 7 (PHP-1, PHP-2, PHP-3) were the liver-damaged rats receiving the PHP at a dose of 50, 100, and 150 mg/kg body weight. Blood samples were collected at 21 of the trial, to evaluate oxidative stress, hepatoprotective and anti-nephrotoxic potential. Results of liver function tests revealed significant (P < .05) hepatoprotective activities of PHP after intoxication with CCl4 of albino rats as compared to standard groups. Moreover, results of renal functions also showed that PHP has a significant (P < .05) restoring the capacity of blood urea, creatinine, and uric acid in intoxicated rats as compared with the control group. The PHP also reduced the oxidative stress in the treatment groups by increasing the total antioxidant capacity and reducing the total oxidative status. It can be concluded that selected medicinal plants have a potential role in the management of liver and kidney disorders. So, by running the clinical trial on a large scale and by isolating the phytochemical constituents responsible for hepatoprotective and nephroprotective activities, locally prepared drugs could be developed to manage liver and renal disorders.

Formulation and characterization of Anacyclus Pyrethrum Emulgels and its in vitro and in vivo evaluation as cosmeceutical product.

BACKGROUND: Plants containing high phenolic and flavonoids contents used widely as antioxidant agent by reducing skin photo damaging effects and play important role in skin rejuvenating. AIMS: This study was performed to explore the cosmetic effects of Anacyclus Pyrethrum extract and to develop stable oil in water (O/W) emulsion base gel loaded with Anacyclus Pyrethrum 10% extract. OBJECTIVE: To explore and quantify phenols and flavonoids present in Anacyclus Pyrethrum extract and determine its cosmetic effects on human skin. METHOD: Emulgel formulation were developed by mixing o/w emulsion with carbopol gelling agent loaded with Anacyclus Pyrethrum (AP) extract and base gel without AP extract. In vitro study was done for the evaluation of color change, liquefaction, hardness, and pH change at different storage condition for the duration of 12 weeks. For in vivo study, emulgel applied on 13 healthy human volunteer's cheeks to evaluate its cosmetics effects and compared with placebo (base). Facial parameters including skin melanin, redness, sebum, moisture content, and skin elasticity were determined by using mexameter, sebumeter, corneometer, elastometer for the study duration of 12 weeks. RESULTS: Total phenolic content in Anacyclus Pyrethrum extract was 80.04 ± 0.0043 mg GAE/g, and flavonoids were 54.64 ± 0.0076 mg QE/g. Anacyclus Pyrethrum extract found significantly effective in reducing skin photo-damage effects (p ≤ 0.05) as compared base gel. CONCLUSION: Anacyclus Pyrethrum extract being rich source of flavonoid and phenolic content, acts as strong antioxidant to protect skin against photo-damaging effect and improve skin conditions.

Anti-inflammatory and antioxidant activity of the hot water-soluble polysaccharides from Anacyclus pyrethrum (L.) Lag. roots.

ETHNOPHAMACOLOGICAL RELEVANCE: the roots of Anacyclus pyrethrum (L.) Lag. (Family: Asteraceae) are used in Algeria to treat respiratory infections, to cure chronic head and nostrils catarrh, and to clear the brain by stimulating the free flow of nasal mucous. They contain a high quantity of hot water-soluble polysaccharides. AIMS OF THE STUDY: The study aims to evaluate the antioxidant and anti-inflammatory activity of polysaccharides extracted from Anacyclus pyrethrum roots (APPS). MATERIALS AND METHODS: The APPS were extracted using boiling water, separated from proteins by the Sevag method then precipitated with 90% ethanol. The antioxidant effect of crude APPS was evaluated using FRAP assay. To investigate the anti-inflammatory potential, mice were treated with crude polysaccharides (25, 50, and 100 mg/kg, i.p.) for 3 days (14th, 15th, and 16th day of the experimentation). Respiratory inflammation was induced by HDM (House Dust Mite), mice were sensitized intranasally with 25 µg HDM suspended in 10 µl NaCl (5 µl/nostril) on days 0 and 7 then challenged with 5 µg HDM on days 14, 15, and 16. Mice were sacrificed 24 h after the last challenge. The number of immune cells in the blood in NL (Nasal Liquid) and in BAL (Broncho Alveolar Liquid) was enumerated, the spleen was removed to calculate the relative spleen weight and to count splénocytes, lungs histopathological examination was carried out to confirm the protective effect of APPS. Structural characterization of APPS was identified using FTIR (Fourier-Transform Infrared Spectroscopy) and SEM (Scanning Electron Microscopy). RESULTS: The crude APPS possessed reducing power. In vivo assay, treatment with APPS causes a decrease in the number of blood leucocytes at all doses on the one hand, and in the relative spleen weight and splénocytes number on the other hand except at the dose of 50 mg/kg in which an enhancement of the number of splénocytes and immune cells in NL and BAL was significant. The histopathological examination showed clear protection of lung tissue damaged by HDM, after treatment with APPS mainly, at the dose of 50 mg/kg. CONCLUSION: Our data clearly showed antioxidant and anti-inflammatory activity of APPS on HDM-challenged mice induced lungs inflammation by equilibrating the inflammatory reaction mostly, with an optimal dose of 50 mg/kg.

Evaluation of the acute toxicity of the extracts of Anacyclus pyrethrum var. pyrethrum (L.) and Anacyclus pyrethrum var. depressus Maire in Swiss mice.

BACKGROUND AND AIM: Anacyclus pyrethrum (L.) has been used in traditional North African and Indian medicine for the treatment of several diseases such as cancer, rheumatism, epilepsy, diabetes, and Alzheimer's disease. Despite its medical benefits, few studies have examined its toxicity. The present study evaluated the acute toxicity of hydroethanolic extracts of different parts (roots, seeds, leaves, and capitula) of two varieties of A. pyrethrum (L.), namely, A. pyrethrum var. pyrethrum (L) and A. pyrethrum var. depressus (Ball) Maire, in mice. MATERIALS AND METHODS: Acute toxicity was evaluated after the oral administration of different extracts at doses of 300, 500, and 2000 mg/kg. Mortality, body weight, general behavior, and adverse effects were observed daily for 14 days. At the end of the experiment, mice were sacrificed, and biochemical parameters and histopathology of the liver, kidneys, and spleen were analyzed. RESULTS: The extracts of different parts of both plants induced no signs of toxicity or mortality during the observation period, excluding capitulum and seed extracts, which induced slight sedation at a dose of 2000 mg/kg. The LD50 of the extracts was estimated to exceed 2000 mg/kg. The administration of A. pyrethrum var. pyrethrum roots at a dose of 300 mg/kg resulted in significantly increased AST levels. However, the A. pyrethrum var. depressus root extract induced significant increases in the levels of both transaminases (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]). The remaining extracts of both plants at a dose of 500 mg/kg significantly increased AST levels. Moreover, all plant extracts excluding the A. pyrethrum var. pyrethrum capitulum extract at 2000 mg/kg provoked significant increases in AST levels, and A. pyrethrum var. depressus roots provoked a significant increase of ALT levels. Meanwhile, mice treated with high doses of extracts (2000 mg/kg) displayed histopathological changes in the liver, kidneys, and spleen characterized by hepatic distress, inflammatory infiltration, focal tubular necrosis, vascular congestion, and lymphoid hyperplasia. CONCLUSION: The results of the present study indicate that the hydroethanolic extracts of different parts of two varieties of A. pyrethrum (L.) were not toxic in mice at low concentrations, whereas some toxic effects were detected in mice treated at 2000 mg/kg.

Anacyclus pyrethrum (L): Chemical Composition, Analgesic, Anti-Inflammatory, and Wound Healing Properties.

BACKGROUND: Anacyclus pyrethrum (A. pyrethrum) is a wild species belonging to the family Asteraceae, which is used in traditional medicines. AIM OF THE STUDY: This work was undertaken to study the chemical composition, analgesic, anti-inflammatory, and wound healing properties of hydroalcoholic extracts of different parts (roots, seeds, leaves, and capitula) of A. pyrethrum. Material and Methods: The phytochemical analysis of the studied extracts was conducted by GC-MS. The analgesic activity was evaluated in mice using acetic acid and formaldehyde methods. The anti-inflammatory activity was tested using the inhibitory method of edema induced in rats. The healing activity of the hydroethanolic extracts was explored by excision and incision wound healing models in rats. RESULTS: The phytochemical analysis of the studied plant extracts affirmed the presence of interesting compounds, including some newly detected elements, such as sarcosine, N-(trifluoroacetyl)-butyl ester, levulinic acid, malonic acid, palmitic acid, morphinan-6-One, 4,5.alpha.-epoxy-3-hydroxy-17-methyl, 2,4-undecadiene-8,10-diyne-N-tyramide, and isovaleric acid. The extracts of different parts (roots, seeds, leaves, and capitula) exhibited promising anti-inflammatory, analgesic, and wound healing effects, with percentages of inhibition up to 98%, 94%, and 100%, respectively. CONCLUSION: This study might contribute towards the well-being of society as it provides evidence on the potential analgesic, anti-inflammatory, and wound healing properties of A. pyrethrum.

Anticonvulsive and neuroprotective effects of aqueous and methanolic extracts of Anacyclus pyrethrum root in kainic acid-induced-status epilepticus in mice.

PURPOSE: Epilepsy is a chronic neurological disorder characterized by spontaneous and recurrent seizures. The currently available synthetic antiepileptic drugs have a limited efficacy and are associated with a wide range of side effects. In Ayurveda, Anacyclus pyrethrum root (APR) has been used as a traditional antiepileptic remedy. The aim of the present study is to evaluate the anticonvulsive and neuroprotective effects of aqueous and methanol extracts of Anacyclus pyrethrum root (AEAPR and MEAPR) on experimental model of status epilepticus (SE). METHODS: Twenty four male mice were divided into four groups. The control and KA groups had free access to tap water for 5 days before the intraperitoneal injection of distillated water or kainic acid (KA; 30 mg/kg), respectively. In the treated groups, mice received extracts solutions MEAPR and AEAPR in drinking water at the concentration of 5 g/l for 5 days. At the fifth day, animals received intraperitoneal injection of KA. The behavioral changes latency of seizures, the number of wet dog shakes (WDS) and the mortality were observed over 6 h. Thereafter, the mice were sacrificed for immunohistochemical studies. RESULTS: Pretreatment with MEAPR and AEAPR decreases significantly the frequency of WDS (32.5% and 43.9%, p < 0.01; respectively), and increases considerably the latent period (77.9% and 91.9%, p<0.01; respectively) between the injection of the KA and the appearance of the SE as compared to the KA group. The duration and severity of seizure in the MEAPR or AEAPR-pretreated groups were significantly lower (p < 0.01 and p < 0.05 or p < 0.01; respectively) than those in the KA group. These behavioral results were confirmed by the immunohistochemical study at the level of the hippocampus, in which the c-FOS and GFAP expression of both MEAPR and AEAPR-treated animals largely reduced (p < 0.001) the number of labelled cells with respect to the group, which received the KA alone. CONCLUSION: Our results showed that the MEAPR and AEAPR have anticonvulsive effect and putative neuroprotective effect against seizures induced by KA. Further studies are required to identify its active ingredients responsible for the observed effects.

Anti-inflammatory, Antinociceptive, and Antioxidant Activities of Methanol and Aqueous Extracts of Anacyclus pyrethrum Roots.

Anacyclus pyrethrum (L.) is a plant widely used in Moroccan traditional medicine to treat inflammatory and painful diseases. The objective of the present study was to evaluate the antinociceptive, anti-inflammatory and antioxidant activities of aqueous and methanol extracts of Anacyclus pyrethrum roots (AEAPR and MEAPR). The anti-inflammatory effect of AEAPR and MEAPR was determined in xylene-induced ear edema and Complete Freund's Adjuvant (CFA)-induced paw edema. The antinociceptive activity of AEAPR and MEAPR (125, 250, and 500 mg/kg) administered by gavage was examined in mice by using acetic acid-induced writhing, hot plate, and formalin tests, and the mechanical allodynia were assessed in CFA-induced paw edema. In addition, the in vitro antioxidant activities of the extracts were determined by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, ferric reducing power and ß-carotene-linoleic acid assay systems. AEAPR and MEAPR produced significant reductions in CFA-induced paw edema and xylene-induced ear edema. A single oral administration of these extracts at 250 and 500 mg/kg significantly reduced mechanical hypersensitivity induced by CFA, which had begun 1 h 30 after the treatment, and was maintained till 7 h. Chronic treatment with both extracts significantly reduced mechanical hypersensitivity in persistent pain conditions induced by CFA. Acute pretreatment with AEAPR or MEAPR at high dose caused a significant decrease in the number of abdominal writhes induced by acetic acid injection (52.2 and 56.7%, respectively), a marked increase of the paw withdrawal latency in the hot plate test, and also a significant inhibition of both phases of the formalin test. This antinociceptive effect was partially reversed by naloxone pretreatment in the hot plate and formalin tests. Additionally, a significant scavenging activity in DPPH, reducing power and protection capacity of ß-carotene was observed in testing antioxidant assays. The present study suggests that AEAPR and MEAPR possess potent anti-inflammatory, antinociceptive and antioxidant effects which could be related to the presence of alkaloids and phenols in the plant. In addition, the antinociceptive effect of APR extracts seems to partly involve the opioid system. Taken together, these results suggest that Anacylcus pyrethrum may indeed be useful in the treatment of pain and inflammatory disorders in humans.

Alkamides from Anacyclus pyrethrum L. and Their in Vitro Antiprotozoal Activity.

In our ongoing study to evaluate the antiprotozoal activity of alkamides from Asteraceae, a dichloromethane extract from the roots of Anacycluspyrethrum L. showed a moderate in vitro activity against the NF54 strain of Plasmodium falciparum and against Leishmaniadonovani (amastigotes, MHOM/ET/67/L82 strain). Seven pure alkamides and a mixture of two further alkamides were isolated by column chromatography followed by preparative high performance liquid chromatography. The alkamides were identified by mass- and NMR-spectroscopic methods as tetradeca-2E,4E-dien-8,10-diynoic acid isobutylamide (anacycline, 1), deca-2E,4E-dienoic acid isobutylamide (pellitorine, 2), deca-2E,4E,9-trienoic acid isobutylamide (3), deca-2E,4E-dienoic acid 2-phenylethylamide (4), undeca-2E,4E-dien-8,10-diynoic acid isopentylamide (5), tetradeca-2E,4E,12Z-trien-8,10-diynoic acid isobutylamide (6), and dodeca-2E,4E-dien acid 4-hydroxy-2-phenylethylamide (7). Two compounds-undeca-2E,4E-dien-8,10-diynoic acid 2-phenylethylamide (8) and deca-2E,4E-dienoic acid 4-hydroxy-2-phenylethylamide (9)-were isolated as an inseparable mixture (1:4). Compounds 3, 4, and 5 were isolated from Anacycluspyrethrum L. for the first time. While compounds 4 and 5 were previously known from the genus Achillea, compound 3 is a new natural product, to the best of our knowledge. All isolated alkamides were tested in vitro for antiprotozoal activity against Plasmodium falciparum, Trypanosomabruceirhodesiense, Trypanosomacruzi, and Leishmaniadonovani and for cytotoxicity against L6 rat skeletal myoblasts.

Anacyclus Pyrethrum Extract Exerts Anticancer Activities on the Human Colorectal Cancer Cell Line (HCT) by Targeting Apoptosis, Metastasis and Cell Cycle Arrest.

INTRODUCTION: Colorectal cancer is the second leading cause of cancer mortality in the USA. There are a number of medicinal plants triggering apoptosis response in cancer cells, thus have a therapeutic potential. On the other hand, due to traditional uses and availability of Anacyclus pyrethrum extract, we decided to evaluate the efficacy of this medicinal herb on human colorectal cancer cell line (HCT). MATERIALS AND METHODS: In the present study, the cytotoxic effects of Anacyclus pyrethrum extract were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and trypan blue viability dye. Then, flow cytometry assay was exploited to measure cell death and apoptosis stage. The scratch test was exploited to assess the effect of Anacyclus pyrethrum on the migration of cancer cells. The expression levels of Caspase 3, Bcl-2, MMP1, and Vimentin genes were quantified by real-time PCR. Finally, cell cycle was analyzed by flow cytometry. RESULTS: MTT assay showed that Anacyclus pyrethrum extract significantly inhibited the cell growth. According to the flow cytometry assay result, the herbal extract was able to induce apoptosis in colorectal cancer cells. Our findings also demonstrated that the plant extract substantially increases the caspase 3 mRNA expression, while decreases Bcl-2, MMP1, and Vimentin. Cell cycle arrest occurred in G1 stage, due to the results of flow cytometry. CONCLUSION: These results indicate that Anacyclus pyrethrum extract can successfully induce apoptosis in HCT cells. Therefore, it could be used as a novel therapeutic candidate for colorectal cancer treatment.

Quantitative transdermal behavior of pellitorine from Anacyclus pyrethrum extract.

The plant Anacyclus pyrethrum (AP) consists of several N-alkylamides with pellitorine as main constituent. AP extracts are known to be biologically active and some products for topical administration containing AP plant extracts are already commercially available with functional cosmeceutical claims. However, no transdermal data for pellitorine are currently available. Therefore, our general goal was to investigate the local skin pharmacokinetics of the plant N-alkylamide pellitorine using a Franz diffusion cell set-up. Two different forms were applied on human skin: purified pellitorine and the AP extract. Our study demonstrated that pellitorine is able to cross the stratum corneum and the subsequent skin layers. A significantly higher permeability coefficient was observed when the AP extract (Kp=2.3 × 10(-4)cm/h) was administered, compared to purified pellitorine (Kp=1.1 × 10(-4)cm/h). With the obtained pellitorine concentrations in the skin layers and the receptor fluid, it is concluded that local and systemic effects can be expected after topical application. Due to these findings and as a regulatory consequence, products containing reasonable concentrations of pellitorine are recommended to be classified as a medicinal product.