The effects of aerial and root extracts on protein phosphorylation were similar in most cases, except for PTEN which was not phosphorylated by root extract and 4E-BP1 which was phosphorylated only by F/extracts increase the phosphorylation of AMPK on thr172. Open in a separate window Fig. and found to be enhanced as well. Glucose uptake was induced by all extracts, with roots found to be the most effective and fruits the least. The effect of on Akt phosphorylation was minor compared to insulin effect. However, GSK3 and PRAS40, which are downstream elements of the insulin cascade, were found to be highly phosphorylated by extracts. Inhibition of PI3K and Akt, but not AMPK and ERK, abrogated the induction of glucose uptake by the aerial parts of the shrub. Conclusion The aerial organs of have anti-diabetic properties and may be used as a basis for the development of dietary supplements or to identify new agents for the treatment of type 2 diabetes. is a widely distributed chamaephyte of the Rosaceae family, growing in the eastern Mediterranean landscape. (aqueous root extract by traditional medicinal practitioners for the treatment of diabetes, as well as for cancer therapy and pain relief [5C10]. In our previous studies, the anti-diabetic properties of root extract of was validated both in-vitro and in-vivo. Root ASP6432 extract facilitated glucose transport into target cells of insulin: hepatocytes, adipocytes, and myotubes. Lipolysis was inhibited by the extract and the signaling pathway of glycogen synthesis was activated [11]. All of these functions suggested that root extract has insulin-mimetic properties. In addition, insulin secretion was enhanced in-vitro, suggesting a stimulatory effect on pancreatic -cells [11]. In-vivo studies revealed that the glucose lowering properties of are accompanied by lower serum insulin levels and improved insulin tolerance [11, 12], indicating that when is administrated to the animal, the improvement in insulin sensitivity enables a reduction, rather than a stimulation, of insulin release. ASP6432 The anti-diabetic activities mentioned in the ethnobotanical literature [5, 6, 13C15], as well as in small experimental studies [16C18], are mostly attributed to the roots of roots. This obstacle is further intensified when taking into account the low seedling rate of [22]. There is limited evidence for the use of the whole plant for the preparation of an antidiabetic remedy rather than only the roots [8, Rabbit Polyclonal to SCN9A 23]. Although activity extracted from a certain plant part suggests the presence of a similar activity from additional plant parts, many examples exist of herbs showing pharmacological properties in one, but not other, organs [24, 25], emphasizing the necessity of experimentally validating the activity of the aerial parts of were investigated for the first time, using several in-vitro models for the study of diabetes. Methods Chemicals, kits and reagents IBMX, dexamethasone, insulin, 2-deoxy-d-glucose (2-DG), cytochalasin-B, -amylase, -glucosidase, dinitrosalicylic acid (DNS), p-nitrophenyl -d-glucopyranoside (PNPG) and inhibitors of proteases and phosphatases were purchased from Sigma. BSA, reagents and media for cell cultures were obtained from Biological Industries (Beit Haemek, Israel). [3H]2-deoxy-d-glucose (1?mCi) and Optiphase scintillation solution were purchased from Perkin-Elmer. A CytoTox 96 assay kit was purchased from Promega. Cytochalasin B, LY294002, AKT inhibitor VIII and Compound C were purchased from CalBiochem. Anti-actin was obtained from MP Biomedicals. Other primary antibodies were obtained from Cell-Signaling Technology. Secondary antibodies were purchased from Jackson ImmunoResearch. Plant material and extract preparation plants were collected from the wild in the area around Ariel University. The plants were identified by the botanical staff of the UniversityA voucher specimen of the plant was deposited in the Israel National Herbarium at the Hebrew University of Jerusalem (No. HUJ 102531). root, leaf, and fruit extracts were prepared according to the traditional method ASP6432 [9]. In 1?L of water, 100?g of fresh root, leaf or fruit (R/or F/extracts. Following 24?h of incubation, LDH was measured in the culture supernatants according to the manufacturers instructions. For positive control, cells were incubated with lysis buffer 60?min prior to measurement. The dye intensity was measured by microplate reader (Tecan, Salzburg, Austria) at a wavelength of 490?nm. -amylase and -glucosidase inhibition assays To determine -amylase inhibition by extracts, the ASP6432 standard procedure [27] was performed. extracts (100?l) were incubated with porcine pancreatic -amylase (250?l, 0.15?U/ml in phosphate buffer (20?mM), pH?6.7, containing 6.7?mM sodium chloride) for 20?min at 37?C. Starch (250?l, 0.5% extracts (25?l) were incubated with -glucosidase (25?l, 0.5?U/ml) for ASP6432 10?min at 37?C, followed by the addition of p-nitrophenyl -d-glucopyranoside.