Violet Kasabri

In vitro and in vivo acute antihyperglycemic effects of five selected indigenous plants from Jordan used in traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE Achillea santolina L., Pistacia atlantica Desf, Rheum ribes L., Sarcopoterium spinosum (L.) Spach and Teucrium polium L. have traditionally been used as herbal antidiabetic medicines. However their alleged benefits and mechanisms remain elusive. AIM OF THE STUDY This study aimed to evaluate the effect of water extracts of these plants in in vitro and in vivo experiments. MATERIALS AND METHODS In vitro enzymatic starch digestion with aqueous extracts from plants at concentrations of 1, 5, 10, 12.5, 25, 50 and 100 mg/ml was assayed using α-amylase and α-amyloglucosidase. Acarbose was used as control and glucose liberation was determined by glucose oxidase method. Oral starch tolerance test (OSTT) and oral glucose tolerance test (OGTT) were determined for the plant extracts at concentrations 125, 250 and 500 mg/kg b.wt. on Sprague-Dawley rats. Blood glucose levels in rats treated with plant extracts and drugs (acarbose or metformin and glipizide) were measured at -30, 0, 45, 90 and 135 min. RESULTS Compared to acarbose (IC(50)=1.2 μg/ml), water extracts of Pistacia atlantica, Rheum ribes and Sarcopoterium spinosum exerted significant dose dependent dual inhibition of α-amylase and α-glucosidase in in vitro enzymatic starch digestion bioassay, with IC(50)s; 46.98, 58.9 and 49.9 mg/ml, respectively. Comparable in vivo results were obtained for starch-fed rats, exhibiting significant acute postprandial antihyperglycemic efficacies. While Achillea santolina and Teucrium polium extracts lacked any favourable in vitro anti-α-amylase and anti-α-glucosidase effect, other modes of action can possibly explain their substantial acute antihyperglycemic activities in starch-treated rats. Except for Pistacia atlantica extracts, none of the investigated extracts qualified for improving the glucose intolerance in fasted rats on glucose loading. CONCLUSIONS Pistacia atlantica, Rheum ribes and Sarcopoterium spinosum can be considered as potential candidates for amelioration/management of type 2 diabetes.

Medicinal Plants from Jordan in the Treatment of Diabetes: Traditional Uses vs. In Vitro and In Vivo Evaluations - Part 2

Diabetes mellitus is the most common metabolic disorder affecting millions worldwide. It is recognized as a global major health problem. As alternatives to the available orthodox medicines, plants are considered a potential source for the treatment of diabetes within traditional ethnomedicine practices. In the Jordanian traditional medicine a significant selection of ethnobotanicals is promoted for their antidiabetic activity. Literature surveys demonstrate the benefit of several ethnobotanicals as antidiabetic agents evaluated in in vitro and in vivo systems in the form of their crude extracts and/or isolated pure compounds with varying degrees of hypoglycemic or antihyperglycemic bioactivities. This mini review discusses the preparatory forms in which these plants are consumed, their reported phytoconstituents, and the results of their reported antidiabetic bioactivity.

Terminalia bellirica stimulates the secretion and action of insulin and inhibits starch digestion and protein glycation in vitro

Traditional plant treatments have been used throughout the world for the therapy of diabetes mellitus. The aim of the present study was to investigate the efficacy and mode of action of Terminalia bellirica used traditionally for the treatment of diabetes in India. T. bellirica aqueous extract stimulated basal insulin output and potentiated glucose-stimulated insulin secretion concentration-dependently in the clonal pancreatic beta-cell line, BRIN-BD11 (P < 0.001). The insulin-secretory activity of the plant extract was abolished in the absence of extracellular Ca2+ and by inhibitors of cellular Ca2+ uptake, diazoxide and verapamil (P < 0.001; n 8). Furthermore, the extract did not increase insulin secretion in depolarised cells and did not further augment insulin secretion triggered by tolbutamide or glibenclamide. T. bellirica extract also displayed insulin-mimetic activity and enhanced insulin-stimulated glucose uptake in 3T3-L1 adipocytes by 300 %. At higher concentrations, the extract also produced a 10-50 % (P < 0.001) decrease in starch digestion in vitro and inhibited protein glycation (P < 0.001). The present study has revealed that components in T. bellirica extract stimulate insulin secretion, enhance insulin action and inhibit both protein glycation and starch digestion. The former actions are dependent on the active principle(s) in the plant being absorbed intact. Future work assessing the use of T. bellirica as a dietary adjunct or as a source of active anti-diabetic agents may provide new opportunities for the treatment of diabetes.

Comparison of the antiproliferative activity of crude ethanol extracts of nine salvia species grown in Jordan against breast cancer cell line models.

Background: The antiproliferative activity of Salvia species grown in Jordan has not been fully evaluated yet. The aim of this work was to study the antiproliferative activity of crude ethanol extracts from nine Salvia species grown in Jordan against a panel of breast cancer cell lines. Material and Methods: Cytotoxic activity was evaluated in human tumor models of breast cancer; MCF-7, T47D, ZR-75-1, and BT 474 by the sulforhodamine B assay. In addition, the extracts were evaluated using a non-transformed cell line (Vero) and normal fibroblast cells in order to demonstrate their selectivity and safety. Results: From the nice ethanol extracts under investigation, those of S. dominica and S. fruticosa showed an inhibitory concentration of 50% of cells (IC50) in concentrations less than 30μg/mL against the four cell lines under investigation. S. syriaca and S. hormium showed an IC50 below 30μg/ml for two out of the four cell lines. S. fruticosa, S. hormium and S. syriaca showed selectivity in their antiproliferative activity against estrogen receptor positive cell lines with minimal toxicity against normal human periodontal fibroblasts. Phytochemical screening using thin layer chromatography indicated the presence of terpenoids, flavonoids and coumarins in all examined extracts. Conclusion: Three of the plant extracts under investigation exhibited antiproliferative activity against breast cancer cells and were shown to be safe and selective. These could be considered as a potential source for novel anticancer therapy.

Evaluation of the acute antihyperglycemic effects of four selected indigenous plants from Jordan used in traditional medicine.

Context: Eryngium creticum Lam. (Umbelliferae), Geranium graveolens L.Her.exn Ait (Geraniaceae), Paronychia argentea Lam. (Caryophyllaceae), and Varthemia iphionoides Boiss (Compositae) have traditionally been used as antidiabetic phytomedicines. However, their alleged benefits and mechanisms remain elusive. Objectives: To evaluate the effect of these plants on in vitro and in vivo enzymatic starch digestion. Materials and methods: In vitro enzymatic starch digestion with acarbose or (1–50 or 100 mg/ml) plants aqueous extracts was assayed using α-amylase and α-amyloglucosidase. Oral starch tolerance tests and oral glucose tolerance tests were determined for the plant extracts at concentrations 125, 250, and 500 mg/kg body weight. Blood glucose levels in rats treated with plant extracts or drugs (acarbose or metformin and glipizide) were measured at −30, 0, 45, 90, and 135 min. Results and discussion: In vitro, acarbose, and water extracts of G. graveolens and V. iphionoides exerted significant dose-dependent dual inhibition of α-amylase and α-glucosidase, with respective IC50s of 1.2 μg/ml, 84.7, and 65.2 mg/ml. Comparable in vivo acute postprandial antihyperglycemic efficacies were obtained for G. graveolens and V. iphionoides in starch-fed rats. E. creticum exhibited substantial acute antihyperglycemic activities in starch-treated rats, despite lacking any favorable in vitro effectiveness. However, P. argentea lacked any inhibitory efficacy. None of the plant extracts qualified for improving the glucose tolerance in fasted rats on glucose loading. Conclusion: G. graveolens and V. iphionoides can be considered as potential candidates for therapeutic modulation of impaired fasting glycemia, impaired glucose tolerance, and type 2 diabetes.

Antiobesity and antihyperglycaemic effects of Adiantum capillus-veneris extracts: in vitro and in vivo evaluations

Abstract Context: Adiantum capillus-veneris L. (Adiantaceae) hypocholesterolemic activity is therapeutically praised. Objectives: Pharmacological modulation of pancreatic triacylglycerol lipase (PL) and α-amylase/α-glucosidase by A. capillus-veneris are evaluated. Materials and methods: Using positive controls (acarbose, orlistat, guar gum, atorvastatin, glipizide and metformin) as appropriate, crude aqueous extracts (AEs) of A. capillus-veneris aerial parts were tested via a combination of in vitro enzymatic (0.24–100 mg/mL), acute in vivo carbohydrate tolerance tests (125, 250 or 500 mg/kg body weight [b.wt]) and chronic in vivo studies (500 mg/kg b.wt) in high cholesterol diet (HCD) fed Wistar rats. Results: Like acarbose, A. capillus-veneris as well as chlorogenic acid, with respective IC50 values (mg/mL) of 0.8 ± 0.0 and 0.2 ± 0.0, were identified as in vitro potent dual inhibitors of α-amylase/α-glucosidase. Unlike guar gum, A. capillus-veneris had no glucose diffusion hindrance capacity. Equivalent to orlistat, A. capillus-veneris and its phytoconstituents inhibited PL in vitro with an ascending order of PL- IC50 values (μg/mL): ferulic acid; 0.48 ± 0.06 < ellagic acid; 13.53 ± 1.83 < chlorogenic acid; 38.4 ± 2.8 < A. capillus-veneris; 1600 ± 100. Incomparable to acarbose or metformin and glipizide, A. capillus-veneris (125, 250 and 500 mg/kg b.wt) lacked antihyperglycaemic efficacies in acute starch- or glucose-evoked postprandial hyperglycaemia increments in normoglycaemic overnight fasting rats. Superior to atorvastatin; A. capillus-veneris exerted significant antiobesity (p < 0.001) with marked triacylglycerol-reducing capacities (p < 0.001) in comparison to rats fed with HCD for 10 weeks. Discussion and conclusion: A. capillus-veneris, modulating pancreatic digestive enzymes, may be advocated as a combinatorial diabesity prevention/phytotherapy agent.

The use of medicinal herbs in gynecological and pregnancy-related disorders by Jordanian women: a review of folkloric practice vs. evidence-based pharmacology

Abstract Context National statistical reports in Jordan indicate a decrease in the total fertility rate along with a parallel increase in contraceptive use. The folkloric use of medicinal herbs in gynecological disorders has been growing in Jordan, despite of deficient reports on the evidence-based safety and efficacy of these practices. Objective The aim of this comprehensive article is to review medicinal plants with claimed ethnonpharmacological usage in various gynecological and pregnancy-related issues in Jordan, and to assess their evidence-based pharmacological studies as well as their phytochemistry. Methods The published literature was surveyed using Google Scholar entering the terms “ethnopharmacology AND Jordan AND infertility AND gynecology OR gestation”. We included ethnopharmacological surveys in Jordan with available full-text. Results Twelve articles were reviewed. Plant species which are commonly used for female gynecological issues such as Artemisia monosperma Del. and A. herba-alba Asso. (Asteraceae) have been found to exert an antifertility effect. Ricinus communis L. (Euphorbiaceae) and Citrullus colocynthis (L.) Schrad. (Cucurbitaceae) had antifertility effects in male rats, but Nigella sativa oil L. (Ranunculaceae) and Cinnamon zeylanicum J. Presl (Lauraceae) were found to enhance it. Conclusion Using plants for gynecological disorders is a common practice in Jordan. Many of them, whether utilised for gynecological or non-gynecological conditions equally, were found to have detrimental effects on female or male fertility. Thus, couples planning pregnancy should be discouraged from the consumption of these herbs. Further local studies are warranted to confirm the appreciable beneficial pharmacological effects and safety of these plants.

Mechanistic studies of antiproliferative effects of Salvia triloba and Salvia dominica (Lamiaceae) on breast cancer cell lines (MCF7 and T47D).

Abstract Ethanol extracts obtained from two Salvia species, S. triloba and S. dominica, collected from the flora of Jordan, were evaluated for their antiproliferative activity against MCF7 and T47D breast cancer cell lines by the sulforhodamine B assay. The ethanol extracts were biologically active with IC50 values of (29.89 ±0.92) and (38.91 ±2.44) μg/mL for S. triloba against MCF7 and T47D cells, respectively, and (5.83 ±0.51) and (12.83 ±0.64) μg/mL for S. dominica against MCF7 and T47D cells, respectively. Flow cytometry analysis and the annexinV-propidium iodide (PI) assay revealed apoptosismediated, and to a lesser extent necrosis-induced, cell death by the S. triloba and S. dominica ethanolic extracts in T47D cells. The mechanism of apoptosis was further investigated by determining the levels of p53, p21/WAF1, FasL (Fas ligand), and sFas (Fas/APO-1). The extract from S. triloba induced a more pronounced enrichment in cytoplasmic mono- and oligonucleosomes than that from S. dominica (p < 0:05) in T47D cells. In response to the extract from S. dominica, but not from S. triloba, the proapoptotic efficacy was specifically regulated by p21. Extracts from both Salvia spp. did not enhance p53 levels, and apoptosis induced by them was not caspase-8- or sFas/FasL-dependent. Thus, our findings indicate that S. triloba and S. dominica ethanolic extracts may be useful in breast cancer management/treatment via proapoptotic cytotoxic mechanisms.

Chemical composition and in vitro studies of the essential oil and aqueous extract of Pelargonium graveolens growing in Jordan for hypoglycaemic and hypolipidemic properties.

Aims: This study aimed to analyze the chemical composition of essential oil of Pelargonium graveolens L. Her. ex Ait. growing in Jordan and to test the efficacy of the leaves aqueous extract and essential oil against pancreatic triacylglycerol lipase (PL), αamylase and α-glucosidase. Study Design: GC-MS analysis of the essential oil obtained by hydrodistillation and Solid Phase Microextraction (SPME) methods as well as in vitro enzymatic investigations. Place and Duration of Study: Faculty of Pharmacy, The University of Jordan, between November 2012 and August 2013. Results: The hydrodistilled oil of P. graveolens fresh leaves yielded twenty eight components, accounting for 95.83 % of the total oil content, while thirty seven components were detected from the fresh leaves by SPME (98.86%). Twenty six and thirty one components were identified in the hydrodistilled and SPME oils of the dried leaves amounting to 96.08 % and 97.83 %, respectively. Oxygenated monoterpenes Original Research Article European Journal of Medicinal Plants, 4(2): , 2014 221 predominated the volatile fractions of the leaves of both methods with citronellol, citronellyl formate and menthone/isomenthone as the major constituents. Similar to orlistat (PL IC50 of 114.0 ± 4.0 ng/mL), P. graveolens extract and volatile oil as well as their purified phyto-constituents inhibited highly substantially in a dose dependent trend PL in vitro (n=3). The P. graveolens extract PLIC50 was 207.4±15.2 μg/mL. As for their volatile oils’ components, PLIC50 (%) (V/V) in an ascending order were: menthone; 0.01±0.0 <geraniol; 0.34±0.02 < linalool; 0.7 ± 0.0 < caryophyllene; 1.17±0.12 <P. graveolens oil; 2.93 ± 0.27. Comparable to acarbose, P. graveolens leaves aqueous extracts (AEs) were identified as in vitro potent and efficacious dual inhibitors of αamylase and α-glucosidase with IC50: 4.6±0.1 mg/mL (p<0.001, n=3). Conclusion: Taken together, P. graveolens leaves, as a nutraceutical modulating gastrointestinal carbohydrate and lipid digestion and absorption, maybe advocated as candidate for obesity-diabetes/metabolic syndrome management.

The correlation between plasma levels of oxytocin and betatrophin in non-diabetic and diabetic metabolic syndrome patients: A cross sectional study from Jordan

BACKGROUND Oxytocin (OXT) is a neurohypophyseal hormone that has been recently shown to possess a number of beneficial effects in diabetes and obesity. Betatrophin is a protein expressed in fat and liver that regulates lipid metabolism and promotes pancreatic β-cell proliferation. It is not investigated yet whether OXT and betatrophin levels correlate in metabolic syndrome (MS) or diabetes patients. METHODS The aim was to assess correlations between plasma betatrophin and OXT levels in MS-diabetic or prediabetic (N=89) as compared to MS-non-diabetic (N=69) patients. Competitive binding ELISA was used to evaluate betatrophin and OXT plasma concentrations. Correlations of the above biomarkers and patient clinical characteristics were also detected. RESULTS As compared to the control MS participants (0.32±0.25ng/mL); betatrophin plasma levels were increased (P<0.001) in the MS-pre/T2DM patients (1.23±0.68ng/mL). On the contrary, OXT concentrations were decreased (P<0.001) in the MS-pre/T2DM patients (1222.46±514.55pg/mL) as compared to the MS control subjects (2323.42±848.68pg/mL). OXT concentration correlated negatively (r=-0.492, P<0.001), while HbA1c and FPG correlated positively with betatrophin plasma levels (P<0.001), but were inversely correlated with OXT levels (P<0.001) in the total sample. CONCLUSION Betatrophin levels are increased, while OXT levels are decreased in MS-pre/T2DM. We found an inverse correlation between the levels of the two biomarkers in addition to correlation between their levels and the degree of glycemic control.