Evaluation of toxicity, nephroprotective and hepatoprotective activities of Argan oil on CCl4-induced nephrotoxicity and hepatotoxicity in Wistar rats

In traditional therapy, Argania spinosa L. seeds oil used as a nephroprotective and hepatoprotective agent. The present work aims to investigate the acute toxicity of unroasted Argan oil, and studied the nephroprotective and the hepatoprotective activity of both oils Roasted (Roil) and unroasted Argan oil (UnRoil) on CCl4-induced liver and kidney damages in Wistar rats. Animals were divided into five equal groups; Control and CCl4 groups are received only distilled water (10 mL/Kg/day). Control positive group received 50 mg/Kg/day of Silymarin. Roil and UnRoil groups treated with 2 mL/Kg/day of Roil and UnRoil. One week after each pretreatment, the rats are injected intraperitoneally with 1 mL/kg/week of CCl4. The treatment has lasted for 15 days. The body weight, urinary volume, water, and food intake were measured at the end of the treatment. Then, the animals are sacrificed; the blood and the liver samples were collected for determining the liver weight ratio and biochemical parameters. UnRoil did not show any sign of toxicity up to 5 mL/Kg. In Roil and UnRoil groups the water intake, ALT, AST, total and direct bilirubin, triglycerides, LDL, plasmatic creatinine, urea, uric acid, and MDA levels are reduced significantly as compared with the CCl4 group. However, body weight,liver weight ratio, food intake, urine urea, urinary creatinine, hepatic glycogen, and GSH levels showed a significant increase compared to the CCl4 group. Roil and UnRoil showed important nephroprotective and hepatoprotective effects against CCl4. Although, the roasting process does not influence the ability of Argan seed oils towards these activities.Keywords

Influence of Abiotic Factors on the Phytochemical Profile of Two Species of Artemisia: A. herba alba Asso and A. mesatlantica Maire

The species of Artemisia are well known in the Mediterranean region, especially in Morocco, for their traditional uses and health benefit. In this study, we were interested in two species of Artemisia, namely A. herba alba Asso and A. mesatlantica Maire. These species were collected from different soils of the Central Middle Atlas (loamy, stony, limestone and rocky soil) with different atmospheres. Extraction of essential oils from the leaves and flowering tops was carried out by hydrodistillation in Clevenger apparatus. Chemical composition analysis was further carried out using gas chromatography coupled with mass spectrometry (GC-MS). Principal component analysis (PCA) was performed to determine the similarities and dissimilarities in the chemical compositions of these six essential oils. The results obtained showed that the essential oil contents extracted from the flowering tops vary from one species to another according to the place of harvest, altitude, soil type and climate. The essential oil yield is between 0.84% and 2.19% (mL/100 g). Chemical analysis revealed that the chemotype of A. herba alba in limestone soil with a subhumid to humid atmosphere is trans-thujone (33.78%), while camphor (46.19%) is for limestone soil with a semi-arid atmosphere, vetivenic acid (14.91%) and davana ether (14.64%) are for limestone soil with a semi-arid and arid atmosphere and camphor (18.39%) is for loamy and stony soil with a semi-arid atmosphere. As for A. mesatlantica from a rocky soil on limestone with a subhumid to humid atmosphere, the main component is camphor (44.86%), and that of limestone soil with a subhumid to the humid atmosphere trans-thujone (41.08%). In addition, HCA affirmed the PCA and allowed us to distinguish between four groups. Our findings observed differences in the chemical compositions of the isolated essential oils most likely related to many factors such as the climates in the regions of the samples collected, altitudes and soil types

Artemisia absinthium L. Aqueous and Ethyl Acetate Extracts: Antioxidant Effect and Potential Activity In Vitro and In Vivo against Pancreatic α-Amylase and Intestinal α-Glucosidase

Artemisia absinthium L. is one of the plants which has been used in folk medicine for many diseases over many centuries. This study aims to analyze the chemical composition of the Artemisia absinthium ethyl acetate and its aqueous extracts and to evaluate their effect on the pancreatic α-amylase enzyme and the intestinal α-glucosidase enzyme. In this study, the total contents of phenolic compounds, flavonoids, and condensed tannins in ethyl acetate and the aqueous extracts of Artemisia absinthium leaves were determined by using spectrophotometric techniques, then the antioxidant capacity of these extracts was examined using three methods, namely, the DPPH (2, 2-diphenyl-1picrylhydrazyl) free radical scavenging method, the iron reduction method FRAP, and the β-carotene bleaching method. The determination of the chemical composition of the extracts was carried out using high-performance liquid chromatography—the photodiode array detector (HPLC-DAD). These extracts were also evaluated for their ability to inhibit the activity of the pancreatic α-amylase enzyme, as well as the intestinal α-glucosidase enzyme, in vitro and in vivo, thus causing the reduction of blood glucose. The results of this study showed that high polyphenol and flavonoid contents were obtained in ethyl acetate extract with values of 60.34 ± 0.43 mg GAE/g and 25.842 ± 0.241 mg QE/g, respectively, compared to the aqueous extract. The results indicated that the aqueous extract had a higher condensed tannin content (3.070 ± 0.022 mg EC/g) than the ethyl acetate extract (0.987 ± 0.078 mg EC/g). Ethyl acetate extract showed good DPPH radical scavenging and iron reduction FRAP activity, with an IC50 of 0.167 ± 0.004 mg/mL and 0.923 ± 0.0283 mg/mL, respectively. The β-carotene test indicated that the aqueous and ethyl acetate extracts were able to delay the decoloration of β-carotene with an inhibition of 48.7% and 48.3%, respectively, which may mean that the extracts have antioxidant activity. HPLC analysis revealed the presence of naringenin and caffeic acid as major products in AQE and EAE, respectively. Indeed, this study showed that the aqueous and ethyl acetate extracts significantly inhibited the pancreatic α-amylase and intestinal α-glucosidase, in vitro. To confirm this result, the inhibitory effect of these plant extracts on the enzymes has been evaluated in vivo. Oral intake of the aqueous extract significantly attenuated starch- and sucrose-induced hyperglycemia in normal rats, and evidently, in STZ-diabetic rats as well. The ethyl acetate extract had no inhibitory activity against the intestinal α-glucosidase enzyme in vivo. The antioxidant and the enzyme inhibitory effects may be related to the presence of naringenin and caffeic acid or their synergistic effect with the other compounds in the extracts

Artemisia absinthium L. Aqueous and Ethyl Acetate Extracts: Antioxidant Effect and Potential Activity In Vitro and In Vivo against Pancreatic α-Amylase and Intestinal α-Glucosidase

Artemisia absinthium L. is one of the plants which has been used in folk medicine for many diseases over many centuries. This study aims to analyze the chemical composition of the Artemisia absinthium ethyl acetate and its aqueous extracts and to evaluate their effect on the pancreatic α-amylase enzyme and the intestinal α-glucosidase enzyme. In this study, the total contents of phenolic compounds, flavonoids, and condensed tannins in ethyl acetate and the aqueous extracts of Artemisia absinthium leaves were determined by using spectrophotometric techniques, then the antioxidant capacity of these extracts was examined using three methods, namely, the DPPH (2, 2-diphenyl-1picrylhydrazyl) free radical scavenging method, the iron reduction method FRAP, and the β-carotene bleaching method. The determination of the chemical composition of the extracts was carried out using high-performance liquid chromatography—the photodiode array detector (HPLC-DAD). These extracts were also evaluated for their ability to inhibit the activity of the pancreatic α-amylase enzyme, as well as the intestinal α-glucosidase enzyme, in vitro and in vivo, thus causing the reduction of blood glucose. The results of this study showed that high polyphenol and flavonoid contents were obtained in ethyl acetate extract with values of 60.34 ± 0.43 mg GAE/g and 25.842 ± 0.241 mg QE/g, respectively, compared to the aqueous extract. The results indicated that the aqueous extract had a higher condensed tannin content (3.070 ± 0.022 mg EC/g) than the ethyl acetate extract (0.987 ± 0.078 mg EC/g). Ethyl acetate extract showed good DPPH radical scavenging and iron reduction FRAP activity, with an IC50 of 0.167 ± 0.004 mg/mL and 0.923 ± 0.0283 mg/mL, respectively. The β-carotene test indicated that the aqueous and ethyl acetate extracts were able to delay the decoloration of β-carotene with an inhibition of 48.7% and 48.3%, respectively, which may mean that the extracts have antioxidant activity. HPLC analysis revealed the presence of naringenin and caffeic acid as major products in AQE and EAE, respectively. Indeed, this study showed that the aqueous and ethyl acetate extracts significantly inhibited the pancreatic α-amylase and intestinal α-glucosidase, in vitro. To confirm this result, the inhibitory effect of these plant extracts on the enzymes has been evaluated in vivo. Oral intake of the aqueous extract significantly attenuated starch- and sucrose-induced hyperglycemia in normal rats, and evidently, in STZ-diabetic rats as well. The ethyl acetate extract had no inhibitory activity against the intestinal α-glucosidase enzyme in vivo. The antioxidant and the enzyme inhibitory effects may be related to the presence of naringenin and caffeic acid or their synergistic effect with the other compounds in the extracts

GATA4 molecular screening and assessment of environmental risk factors in a Moroccan cohort with tetralogy of Fallot

Background: Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart defect (CHD) with an incidence of 1/3600 live births. This disorder was associated with mutations in the transcription factors involved in cardiogenesis, like Nk2 homeobox5 (NKX2-5), GATA binding protein4 (GATA4) and T-BOX1 (TBX1). GATA4 contributes particularly to heart looping and differentiation of the second heart field.Objectives: The aim of this study was to screen a Moroccan cohort with tetralogy of Fallot for GATA4 mutations, and to assess environmental risk factors that could be involved in the occurrence of this disorder.Methods: Thirty-one non-syndromic TOF patients, enrolled between 5th April 2014 and 18th June 2015, were screened for GATA4 mutations using direct sequencing of GATA4 coding exons. Statistical assessment of different risk factors, which is a retrospective study, was carried out using Chi-square and Fisher’s exact tests. Results: We identified seven exonic variants in nine patients (two missense and five synonymous variants); in addition of eight intronic variants. Assessment of environmental risk factors shows significant association of maternal passive smoking with TOF in the Moroccan population.Conclusion: The present study allowed, for the first time, the molecular and environmental characterisation of Moroccan TOF population. Our findings emphasise particularly the strong association of passive smoking with the emergence of tetralogy of Fallot.Keywords: Tetralogy of Fallot, GATA4, molecular screening, risk factors

Cedrus atlantica (Endl.) Manetti ex Carrière Essential Oil Alleviates Pain and Inflammation with No Toxicity in Rodent

Cedrus atlantica (Endl.) Manetti ex Carrière is an endemic tree with spiritual value, and it was used since immemorial time in folk medicine. The present study aims to evaluate the anti-inflammatory (carrageenan-induced paw edema and formalin tests) and analgesic effects (hot plate and acetic acid writhing tests) of the cedarwood essential oil, as well as inspect any toxicity (acute toxicity), using several in vivo assays. Following the acetic acid writhing test and the hot plate test, the EO presented an excellent analgesic effect compared to the controls, especially with the dose of 50 mg/kg. Similar results were found while assessing the anti-inflammatory potential in the carrageenan-induced paw edema and formalin assays. The acute toxicity assessment and the subsequent monitoring of the animals, the biochemical analysis, and the relative organ weight, demonstrated a total safety of the EO. The GC/MS analysis of the composition revealed that the major compounds contained in this EO are beta-himachalene (51.95%), followed by alpha-himachalene (15.82%), and gamma-himachalene (12.15%). This study supports the usage of this tree EO to alleviate pain and inflammation

Influence of Abiotic Factors on the Phytochemical Profile of Two Species of Artemisia : A. herba alba Asso and A. mesatlantica Maire

The species of Artemisia are well known in the Mediterranean region, especially in Morocco, for their traditional uses and health benefit. In this study, we were interested in two species of Artemisia, namely A. herba alba Asso and A. mesatlantica Maire. These species were collected from different soils of the Central Middle Atlas (loamy, stony, limestone and rocky soil) with different atmospheres. Extraction of essential oils from the leaves and flowering tops was carried out by hydrodistillation in Clevenger apparatus. Chemical composition analysis was further carried out using gas chromatography coupled with mass spectrometry (GC-MS). Principal component analysis (PCA) was performed to determine the similarities and dissimilarities in the chemical compositions of these six essential oils. The results obtained showed that the essential oil contents extracted from the flowering tops vary from one species to another according to the place of harvest, altitude, soil type and climate. The essential oil yield is between 0.84% and 2.19% (mL/100 g). Chemical analysis revealed that the chemotype of A. herba alba in limestone soil with a subhumid to humid atmosphere is trans-thujone (33.78%), while camphor (46.19%) is for limestone soil with a semi-arid atmosphere, vetivenic acid (14.91%) and davana ether (14.64%) are for limestone soil with a semi-arid and arid atmosphere and camphor (18.39%) is for loamy and stony soil with a semi-arid atmosphere. As for A. mesatlantica from a rocky soil on limestone with a subhumid to humid atmosphere, the main component is camphor (44.86%), and that of limestone soil with a subhumid to the humid atmosphere trans-thujone (41.08%). In addition, HCA affirmed the PCA and allowed us to distinguish between four groups. Our findings observed differences in the chemical compositions of the isolated essential oils most likely related to many factors such as the climates in the regions of the samples collected, altitudes and soil typesPeer reviewe

Phytochemical Analysis, Antimicrobial and Antioxidant Properties of Thymus zygis L. and Thymus willdenowii Boiss. Essential Oils

Essential oils (EOs) are chemical products produced by odoriferous glands from a variety of plants. These essential oils have many health benefits: antiseptic, anti-inflammatory and antimicrobial activities. So due to these medicinal properties, the present study was designed to analyze essential oils of Thymus zygis L. and Thymus willdenowii Boiss. for their chemical composition and biological activities. These two thyme species were collected from the region of Ifrane, Middle Atlas of Morocco. The EO was obtained by hydrodistillation, and the yields were 5.25% for T. zygis and 3.00% for T. willdenowii. The chemical composition of the EOs was analyzed by gas chromatography coupled with mass spectrometry (GC-MS), and the results showed that T. zygis EO is dominated by carvacrol (52.5%), o-cymene (23.14%), and thymol (9.68%), while the EO of T. willdenowii contains germacrene D (16.51%), carvacrol (16.19%), and geranyl acetate (8.35%) as major compounds. The antioxidant activity assessed by Diphenylpicrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays revealed that both EOs have excellent antioxidant activities; by DPPH it resulted in IC50 = 6.13 ± 0.11 for T. zygis and 6.78 ± 0.3 µg/mL for T. willdenowii, while the one by FRAP yielded EC50 = 2.46 ± 0.01 (T. zygis) and 5.17 ± 0.2 (T. willdenowii) µg/mL. The antimicrobial activity of the two essential oils was evaluated against six bacterial strains and five fungal strains by the disk diffusion method to determine the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC). The EOs revealed variable antimicrobial activities against the different tested microbial strains and showed strong antimicrobial activities, even against strains known as multi-resistant to antibiotics (Acinetobacter baumannii) at low concentrations (2 µL/mL). T. zygis EO showed the most powerful activity against all the studied bacteria, while that of T. willdenowii recorded moderate activity when tested against Shigella dysenteriae and Salmonella Typhi. With inhibition diameters that vary between 75 mm and 84 mm for concentrations of 2 µL/mL up to 12 µL/mL, S. aureus was shown to be the most sensitive to T. zygis EO. For the antifungal activity test, T. zygis EO showed the best inhibition diameters compared to T. willdenowii EO. These results showed that T. zygis EO has more powerful antioxidant and antimicrobial activities than T. willdenowii EO, therefore, we deduce that thyme EOs are excellent antioxidants, they have strong antimicrobial properties, and may in the future represent new sources of natural antiseptics that can be used in pharmaceutical and food industry

Chemical Composition, Antioxidant and Antibacterial Activities of Thymus broussonetii Boiss and Thymus capitatus (L.) Hoffmann and Link Essential Oils

Thymus capitatus and Thymus broussonnetii are two Moroccan endemic medicinal plants used traditionally by the local population. The present study aims to investigate their essential oil chemical composition, antioxidant and antibacterial activities. The chemical composition of the essential oils was determined using the GC-MS analysis, the antioxidant activity assessed using DPPH and FRAP methods while the antimicrobial activity was evaluated against nine bacteria species tested (Enterococcus faecalis, Serratia fonticola, Acinetobacter baumannii, Klebsiella oxytoca, sensitive Klebsiella pneumoniae, sensitive Escherichia coli, resistant Escherichia coli, resistant Staphylococcus aureus and Enterobacter aerogenes). The major identified compounds of T. capitatus essential oil where carvacrol (75%) and p-cymene (10.58%) while carvacrol (60.79%), thymol (12.9%), p-cymene (6.21%) and γ-terpinene (4.47%) are the main compounds in T. broussonnetii essential oil. The bioactivity of the essential oils of the two species of thyme was explained by their richness in oxygenated monoterpenes known for their great effectiveness with an IC50 of 3.48 ± 0.05 and 4.88 ± 0.04 μL/mL and EC50 of 0.12 ± 0.01 and 0.20 ± 0.02 μL/mL in the DPPH and FRAP assays, respectively, with an important antibacterial activity. These results encourage the use of these plants as a source of natural antioxidants, and antibacterial additives, to protect food from oxidative damage and to eliminate bacteria that are responsible for nosocomial infections