Bioactive compounds from Hypericum humifusum and Hypericum perfoliatum: inhibition potential of polyphenols with acetylcholinesterase and key enzymes linked to type-2 diabetes.

CONTEXT: Natural products are reported to have a wide spectrum of pharmacological properties such as antimicrobial, anti-inflammatory and anti-cholinesterase. The genus Hypericum (Hypericaceae) is a source of a variety of molecules with different biological activities, notably hypericin and various phenolics. OBJECTIVES: The goals of the present work were the determination of total phenolic and flavonoid content, hypericin and hyperforin concentration as well as the evaluation of biological of Hypericum humifusum L. (Hhu) and Hypericum perfoliatum L. (Hper). MATERIALS AND METHODS: The various extracts of aerial parts were powdered, and then extracted with methanol. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) and minimum bactericidal (MBC) methods against four Gram-positive bacteria, four Gram-negative bacteria and yeast. RESULTS: The results revealed that H. humifusum, bear the highest total phenolic and flavonoid content (48-113 mg GAE/g and 8-41 mg RE/g, respectively) as well as hypericin (60-90 mg/g) and hyperforin (8-30 mg/g) concentration. Both species showed significant antioxidant activity as revealed by DPPH, FRAP, ABTS, and metal chelating assays. H. humifusum exhibited a strong acetylcholinesterase (3.86-4.57 mg GALAEs/g), α-glucosidase (0.73-2.55 mmol ACEs/g) and α-amylase (3-8 mmol ACEs/g) inhibitory activity. The extract of H. humifusum exhibited strong antibacterial activity mainly against Staphylococcus epidermidis, Staphylococus aureus, and Enterococcus faecium (MIC values ranging from 200 to 250 µg/mL). The highest antifungal activity was showed for H. perfoliatum extract (MIC value = 250 µg/mL). CONCLUSION: The data suggest that H. humifusum could be used as valuable new natural agents with functional properties for pharmacology industries.

Lycium intricatum Boiss.: An unexploited and rich source of unsaturated fatty acids, 4-desmethylsterols and other valuable phytochemicals.

BACKGROUND: Lycium intricatum Boiss., a Solanaceous shrubbery is used in Tunisia as a windbreak and medicinal plant. However, it is considered as underexploited specie despite its high potential to serve as source with economic and nutritional value. To date only limited information about its phytochemistry, especially of its oil has been published. This work provides data on fatty acids, phytosterols and vitamin D composition of L. intricatum seed oil. It opens up new possibilities of developing L. intricatum as a new crop that contains phytochemicals with high added value little influenced by selection or commercial breeding. FINDINGS: The composition of fatty acids, phytosterols and vitamin D in L. intricatum seed oil was assessed by GC-FID. The main fatty acids of L. intricatum seed oil were linoleic acid (49.47%), palmitoleic acid (27.96%) and erucic acid (13.62%). Palimtic acid was present at low percentage (0.63%). The content of unsaturated fatty acids was high as 94.04%. The sterolic fraction was composed of stigmasterol (18.56 mg/100 g), ß-sitosterol (13.04 mg/100 g). L. intricatum oil is an oily matrix that contains hydrocarbons, mainly squalene (63.36 mg/100 g), and two triterpenic alcohol erythrodiol (80.36 mg/100 g) and uvaol (24.06 mg/100 g). provitamin D was present in high quantity (8.12 mg/100 g). CONCLUSIONS: From these results it has been shown that L. intricatum seeds have great potential as a source of fatty acids and phytosterols for natural health products.

Essential oil composition and antibacterial activity of Origanum vulgare subsp. glandulosum Desf. at different phenological stages.

Variation in the quantity and quality of the essential oil (EO) of wild population of Origanum vulgare at different phenological stages, including vegetative, late vegetative, and flowering set, is reported. The oils of air-dried samples were obtained by hydrodistillation. The yield of oils (w/w%) at different stages were in the order of late vegetative (2.0%), early vegetative (1.7%), and flowering (0.6%) set. The oils were analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). In total, 36, 33, and 16 components were identified and quantified in vegetative, late vegetative, and flowering set, representing 94.47%, 95.91%, and 99.62% of the oil, respectively. Carvacrol was the major compound in all samples. The ranges of major constituents were as follows: carvacrol (61.08-83.37%), p-cymene (3.02-9.87%), and γ-terpinene (4.13-6.34%). Antibacterial activity of the oils was tested against three Gram-positive and two Gram-negative bacteria by the disc diffusion method and determining their diameter of inhibition and the minimum inhibitory concentration (MIC) values. The inhibition zones and MIC values for bacterial strains, which were sensitive to the EO of O. vulgare subsp. glandulosum, were in the range of 9-36 mm and 125-600 µg/mL, respectively. The oils of various phenological stages showed high activity against all tested bacteria, of which Bacillus subtilis was the most sensitive and resistant strain, respectively. Thus, they represent an inexpensive source of natural antibacterial substances that exhibited potential for use in pathogenic systems.

Variation of the chemical composition of essential oils in Tunisian populations of Thymus algeriensis Boiss. et Reut. (Lamiaceae) and implication for conservation.

The variation of the essential-oil composition among 14 Tunisian natural populations of Thymus algeriensis Boiss. et Reut. (=Thymus hirtus Willd. ssp. algeriensis Boiss. et Reut.) was assessed by GC (RI) and GC/MS. The populations were collected from different geographical regions belonging to the sub-humid, upper semi-arid, mean semi-arid, lower semi-arid, and upper-arid bioclimates. A total of 47 constituents, representing 81.0 to 96.5% of the total oil, were identified. The main volatiles at the species level were 1,8-cineole (17.7%), alpha-pinene (15.5%), and camphor (8.2%). A high variation among populations for the majority of the compounds was shown. Camphor (0.2-14.0%), linalool (0.2-22.4%), borneol (<0.01-24.3%), caryophyllene oxide (<0.01-18.8%), thymol (<0.01-54.9%), gamma-terpinene (0.4-6.5%), alpha-copaene (0.4-7.6%), linalyl acetate (<0.01-6.4%), and methyl eugenol (<0.01-6.9%) were the main constituents differentiating the populations. The chemical differentiation among populations, assessed by principal component analysis (PCA) and a UPGMA (unweighted pair-group method with averaging) cluster analysis performed on all populations and compounds, was high. Six chemotypes according to the main compounds have been distinguished, i.e., caryophyllene oxide/1,8-cineole/alpha-pinene, 1,8-cineole/alpha-pinene, 1,8-cineole/alpha-pinene/camphor, borneol/1,8-cineole/alpha-pinene, linalool, and thymol chemotypes. The spatial chemotype distribution was linked to the geographic distance among populations rather than to bioclimates, indicating that local selective environmental factors act on the chemotype diversity. The high chemical variation among populations according to their geographical and bioclimatic distribution imposes that conservation strategies of populations should be made appropriately, taking into account these factors. The in situ and ex situ conservation strategies should concern all populations representing the different chemotypes.

Genetic diversity and population structure of Teucrium polium (Lamiaceae) in Tunisia.

Random amplified polymorphic DNA markers were used to assess the genetic diversity within and among seven Tunisian diploid and polyploid populations of Teucrium polium L. from five bioclimatic areas. Out of the 141 bands generated from eight selected primers, 124 were polymorphic. The genetic diversity within a population (Shannon's index) was high and varied according both the ploidal levels and bioclimatic zones. The genetic differentiation among populations assessed by G (ST) and Phi(ST) statistics was high, suggesting a low level of gene flow among them. The major proportion of the variation was attributable to individual differences within populations. The UPGMA analysis based on Nei and Li's coefficient showed that individuals from each population clustered together. In a dendrogram using the Phi(ST) distance matrix, population grouping is concordant with bioclimates and cytotypes. Conservation strategies should take into account the level of the genetic diversity of the populations according to their bioclimate and ploidal levels.

Variation of volatiles in Tunisian populations of Teucrium polium L. (Lamiaceae).

Volatiles from wild Tunisian diploid (2n=2x=26) and tetraploid (2n=4x=52) Teucrium polium L. populations from five bioclimatic zones were assessed by GC and GC/MS. Thirty-eight compounds were identified. Main volatiles at the species level were myrcene (6; 15.3%), germacrene D (27; 9.0%), alpha-pinene (6.6%), beta-pinene (3; 5.8%), and alpha-cadinol (38; 5.1%). A significant variation among populations was observed. The tetraploid populations belonging to the sub humid and lower semi arid bioclimates showed a high amount of myrcene (6; 24.2-43.8%), those from the upper arid contain high percentages of alpha-cadinol (38; 50.6%). The dominant compounds in diploid populations, mainly located in the semi arid zone, are alpha-pinene (1) and germacrene D (27). Population chemical structure, assessed by a principal component analysis (PCA) using all identified compounds, is concordant with bioclimate and ploidy level. In situ conservation of populations should be made appropriately according to these two factors.