DNA Barcoding of Two Thymelaeaceae Species: Daphne mucronata Royle and Thymelaea hirsuta (L.) Endl

Daphne mucronata Royle and Thymelaea hirsuta (L.) Endl both belong to the Thymelaeaceae family. Both species are used traditionally to treat several diseases along with various daily applications by Jordanian Bedouins. Traditionally, those species are identified through personal proficiency, which could be misleading due to human errors or lack of expertise. This study aims to investigate an effective DNA barcoding method to identify and characterize Daphne mucronata Royle and Thymelaea hirsuta plant species at the molecular level. Daphne mucronata Royle and Thymelaea hirsuta were collected from the ancient city of Petra in the Southern part of Jordan. Sequences of candidate DNA barcodes were amplified (rbcL, matK, and rpoC1), sequenced, and aligned to the blastn database. Moreover, the obtained sequences were compared with available sequences of related species at the GenBank database. Our results showed that DNA barcoding successfully identifies the two plant species using any of chloroplast genes (rbcL, matK, or rpoC1). The results emphasize the ability of DNA barcoding for identifying and characterizing different plant species through the recruitment of different barcode loci in molecular identification.

Beneficial Effect of Thymelaea hirsuta on Pancreatic Islet Degeneration, Renal Fibrosis, and Liver Damages as Demonstrated in Streptozotocin-Induced Diabetic Rat

Objective. In Morocco, Thymelaea hirsuta (T. hirsuta) (Thymelaeacea) is a medicinal plant widely used to treat and prevent diabetes. The present study aimed to evaluate the medium-term antidiabetic effect of aqueous extract (AqTh) and ethyl acetate fraction (EaTh) of Th and to investigate their putative protective effect on pancreatic islet degeneration, diabetic nephropathy, and liver damages in streptozotocin (STZ)-diabetic rats. Methods. Experimental diabetes in rats was induced by a single intraperitoneal injection of 50 mg/kg of STZ. During the treatment period (4 weeks), 200 mg/kg AqTh and 50 mg/kg EaTh were orally administrated daily to STZ-diabetic rats. A group of parameters including fasting blood glucose, biochemical parameters, and intestinal α-glucosidase inhibition were studied. Furthermore, histological study of the pancreas, kidney, liver, and aorta was also realized. Results. At the end of the treatment, both AqTh and EaTh had normalized fasting blood glucose to 1.08 and 1.25 g/l, respectively. AqTh has also reduced urinary creatinine and HbAc1. The EaTh showed inhibitory activity against intestinal α-glucosidase, whereas AqTh did not have this inhibitory effect. Furthermore, pancreas hematoxylin and eosin staining showed that AqTh or EaTh prevents pancreatic islet cell degeneration. As the same kidney, Masson’s trichrome staining has shown a significant prevention of renal fibrosis in AqTh- or EaTh-treated diabetic rats. On the other hand, liver hematoxylin and eosin staining showed that AqTh and EaTh prevent liver damage. Conclusion. We conclude that medium-term administration of AqTh and EaTh exerts significant antihyperglycemic effect in STZ-diabetic rats possibly through intestinal α-glucosidase inhibition and protection against pancreatic islet cell damage. Moreover, AqTh and EaTh treatment prevent nephropathy and liver complications in STZ-diabetic rats.

Antifungal and Antiaging Evaluation of Aerial Part Extracts of Thymelaea hirsuta (L.) Endl

In western Algeria, Thymelaea hirsuta aerial parts are traditionally used topically for the treatment of human skin infections. For a better knowledge of its biological activities, this study aimed at evaluating the in vitro antifungal activity and antiaging properties of several extracts of the aerial parts by in vitro assays. Antifungal activity was investigated by using the reference microbroth dilution method (National Committee for Clinical Laboratory Standards) against the dermatophytes ( Microsporum audouinii, Nannizzia gypsea, Trichophyton interdigitale, and Trichophyton rubrum) and the yeast ( Candida albicans). Antiaging (inhibition of 1,1-diphenyl-2-picrylhyorazyl radical scavenging and tyrosinase, lipoxygenase, elastase, and hyaluronidase activities) assays were performed in 96-well plates. All the extracts of T. hirsuta exhibited antifungal activity against M. audouinii, T. rubrum, and C. albicans, while no activity was found against N. gypsea and T. interdigitale. The highest activity was observed on M. audouinii for the aqueous leaf (minimum inhibitory concentration [MIC] of 8 µg/mL) and stem extracts (MIC = 31 µg/mL) and the methanolic leaf extract (MIC = 8 µg/mL). Additionally, some marked morphological alterations were observed on M. audouinii hyphae with all aqueous extracts. Concerning antiage bioassays, the major activity was found against hyaluronidase for the aqueous and methanolic extracts from leaves and the methanolic extract from flowers. Analysis of this last extract by high-performance liquid chromatography, electrospray ionization-mass spectrometry, and nuclear magnetic resonance, allowed us to identify daphnoretin and daphnetin as major components of this extract, and, therefore, responsible for the antiaging activity. In conclusion, this study provides additional experimental data supporting the traditional use of T. hirsuta extracts on some cutaneous dermatophytosis, as well as new perspectives on the potential use of T. hirsuta to ensure better healing of the skin.

New Antiproliferative Triflavanone from Thymelaea hirsuta—Isolation, Structure Elucidation and Molecular Docking Studies

In this study isolates from Thymelaea hirsuta, a wild plant from the Sinai Peninsula of Egypt, were identified and their selective cytotoxicity levels were evaluated. Phytochemical examination of the ethyl acetate (EtOAc) fraction of the methanolic (MeOH) extract of the plant led to the isolation of a new triflavanone compound (1), in addition to the isolation of nine previously reported compounds. These included five dicoumarinyl ethers found in Thymelaea: daphnoretin methyl ether (2), rutamontine (3), neodaphnoretin (4), acetyldaphnoretin (5), and edgeworthin (6); two flavonoids: genkwanin (7) and trans-tiliroside (8); p-hydroxy benzoic acid (9) and β sitosterol glucoside (10). Eight of the isolated compounds were tested for in vitro cytotoxicity against Vero and HepG2 cell lines using a sulforhodamine-B (SRB) assay. Compounds 1, 2 and 5 exhibited remarkable cytotoxic activities against HepG2 cells, with IC50 values of 8.6, 12.3 and 9.4 μM, respectively, yet these compounds exhibited non-toxic activities against the Vero cells. Additionally, compound 1 further exhibited promising cytotoxic activity against both MCF-7 and HCT-116 cells, with IC50 values of 4.26 and 9.6 μM, respectively. Compound 1 significantly stimulated apoptotic breast cancer cell death, resulting in a 14.97-fold increase and arresting 40.57% of the cell population at the Pre-G1 stage of the cell cycle. Finally, its apoptosis-inducing activity was further validated through activation of BAX and caspase-9, and inhibition of BCL2 levels. In silico molecular docking experiments revealed a good binding mode profile of the isolates towards Ras activation/pathway mitogen-activated protein kinase (Ras/MAPK); a common molecular pathway in the development and progression of liver tumors.

Genetic and Biochemical Studies of Thymelaea hirsuta L. Growing Naturally at the North Western Coast of Egypt

This study aims to elucidate the biochemical and genetic behaviors of Thymelaea hirsuta L. growing naturally at the following habitats; ooletic sand dunes, slope of salt marshes, non-saline depression, road side, plateau, upstream of Wadi Halazeen, and wadi bed of Wadi Halazeen along the western Mediterranean coast of Egypt. Soil and plant samples were collected in May, 2017. Water content recorded the highest value in Thymelaea growing at slope of salt marshes. Total soluble carbohydrate attained the highest value in Thymelaea growing at up-stream of Wadi Halazeen while the highest values in none-souble carbohydrate and proline were observed in plants growing at ooletic sand dunes. Total carbohydrate recorded the highest value in Thymelaea growing at plateaue. Total lipids and total phenols attained the highest content in Thymelaea growing at wadi bed. Genetically, ISSR & SRAP molecular markers were done. Seven ISSR and four SRAP primes produced 73.78% and 68.57% of polymorphism, respectively. PIC value, assay efficiency index, effective multiples ratio and marker index, were higher in SRAP than in ISSR.

Notas sobre Thymelaea hirsuta (L.) Endl. y Thymelaea lythroides Barratte & Murb. en la provincia de Sevilla (España)

Thymelaea lythroides es una especie amenazada de la que se conocen sólo dos poblaciones en la Península Ibérica. Se localiza una nueva población de la que se aportan datos sobre la vegetación circundante, la distribución espacial y la relación de sexos de los individuos que la conforman. También se aportan dos inventarios con Thymelaea hirsuta y se discute la serie de vegetación en la que aparece esta planta que resulta novedad para la provincia de Sevilla.

Notas sobre Thymelaea hirsuta (L.) Endl. y Thymelaea lythroides Barratte & Murb. en la provincia de Sevilla (España)

Thymelaea lythroides es una especie amenazada de la que se conocen sólo dos poblaciones en la Península Ibérica. Se localiza una nueva población de la que se aportan datos sobre la vegetación circundante, la distribución espacial y la relación de sexos de los individuos que la conforman. También se aportan dos inventarios con Thymelaea hirsuta y se discute la serie de vegetación en la que aparece esta planta que resulta novedad para la provincia de Sevilla.

Polyphenols contents and antioxidant Activity of extracts from Leaves and flowers of Thymelaea hirsuta

Thymelaea hirsuta is a medicinal plant, belonging to the genus Thymelaea (Thymelaeaceae) widely used in Mediterranean countries especially in Algeria. In this study, we have evaluated the total polyphenols and flavonoids contents of methanolic and aqueous extracts of Leaves and flowers of Thymelaea hirsuta as well as its antioxidant activity using the DPPH (2'2-diphenylpicrylhydrazyl) and β-carotene /linoleic acid bleaching assays. The yield of the methanolic and aqueous extract was 11, 55% and 13, 25% respectively. The total polyphenols content of the methanolic extract was 295, 22 µg GAE/ mg extract and flavonoids was 09, 40 µg QE/mg and 26, 42 µg RE/ mg extract. The total polyphenols content of the aqueous extract was 57, 95 µg GAE/ mg extract and flavonoids was 4.59 µg QE/ mg and 10.66 µg RE/ mg extract. In the DPPH assay, methanolic extract showed the higher scavenging capacity (IC50 = 0.03 ± 0.004 mg/ml), followed by aqueous extract with IC50 of 0.275 ± 0.019 mg/ml. In the test of β-carotene /linoleic acid, the percentage of inhibition was 39.1±1.33% for the aqueous extract and 41.05±2.72% for methanolic extract. Keywords: Thymelaea hirsuta, antioxidant activity, polyphenols, DPPH scavenging, β-carotene.

Acute Toxicity Study of Aqueous Extracts of Thymelaea hirsuta Leaves And Flowers in Rabbits

Thymelaea hirsuta is a medicinal plant known for its therapeutic virtues in traditional medicine. The objective of this study was to evaluate the acute toxicity of aqueous extract of leaves and flowers of Thymelaea hirsuta in rabbit. Aqueous extract was administered to rabbit in single doses of 2.5, 4, 5.5 and 7 g/kg given by gavage. General behavior, adverse effects and mortality were determined for up to 7 days. Biochemical parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), cholesterol (CHOL) and glucose (GLU) were determined after 7 days. The results showed no mortality was observed during the 7 days of the observation period, an increase in AST and ALT in treated groups at the doses of 4, 5.5 and 7 g/kg, and a decrease in both cholesterol and glucose levels in treated groups compared to control. Keywords: Thymelaea hirsuta, Acute toxicity, Biochemical parameters, adverses effects.