Comparative Analysis of Chemical Profile, Antioxidant, In-vitro and In-vivo Antidiabetic Activities of Juniperus foetidissima Willd. and Juniperus sabina L.

Fruit and leaves of junipers are commonly used internally as tea and pounded fruits are eaten to lower blood glucose levels in Anatolia. Thus, we aimed to evaluate antidiabetic and antioxidant potential and the chemical profile of Juniperus foetidissima Willd. and J. sabina L. in this study. In-vitro antidiabetic activities of leaf and fruit extracts were examined by their inhibitory activity on α-glucosidase and α-amylase enzymes. Then, in-vivo antidiabetic activities of leaf and fruit extracts of Juniperus species were investigated on streptozotocin-induced diabetic rats. Additionally, antioxidant activities (phosphomolybdenum, ferric-reducing antioxidant power and ABTS radical scavenging activity assays), phytochemical screening tests and high performance liquid chromatography analysis (HPLC) were done. In-vitro enzyme inhibitory effects of the extracts were supported by the results of in-vivo antidiabetic activity studies. Phytochemical screening tests indicated presence of flavonoids, tannins, terpenoids and carbohydrates in the extracts. Amentoflavone was identified as the major compound in the extracts and content of amentoflavone was determined. As a result, Juniperus extracts and its active constituents might be beneficial for diabetes and its complications.

Enzyme inhibitory and radical scavenging effects of some antidiabetic plants of Turkey.

OBJECTIVES: Ethnopharmacological field surveys demonstrated that many plants, such as Gentiana olivieri, Helichrysum graveolens, Helichrysum plicatum ssp. plicatum, Juniperus oxycedrus ssp. oxycedrus, Juniperus communis var. saxatilis, Viscum album (ssp. album, ssp. austriacum), are used as traditional medicine for diabetes in different regions of Anatolia. The present study was designed to evaluate the in vitro antidiabetic effects of some selected plants, tested in animal models recently. MATERIALS AND METHODS: α-glucosidase and α-amylase enzyme inhibitory effects of the plant extracts were investigated and Acarbose was used as a reference drug. Additionally, radical scavenging capacities were determined using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) ABTS radical cation scavenging assay and total phenolic content of the extracts were evaluated using Folin Ciocalteu method. RESULTS: H. graveolens ethanol extract exhibited the highest inhibitory activity (55.7 % ± 2.2) on α-amylase enzyme. Additionally, J. oxycedrus hydro-alcoholic leaf extract had potent α-amylase inhibitory effect, while the hydro-alcoholic extract of J. communis fruit showed the highest α-glucosidase inhibitory activity (IC50: 4.4 µg/ml). CONCLUSION: Results indicated that, antidiabetic effect of hydro-alcoholic extracts of H. graveolens capitulums, J. communis fruit and J. oxycedrus leaf might arise from inhibition of digestive enzymes.

A bioactivity guided study on the antidiabetic activity of Juniperus oxycedrus subsp. oxycedrus L. leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Juniperus (Cupressaceae) species are widely used as folk medicine in spreading countries. Decoction of Juniperus oxycedrus subsp. oxycedrus L. leaves is used internally to lower blood glucose levels in Turkey. AIM OF THE STUDY: To determine hypoglycaemic and antidiabetic activities of Juniperus oxycedrus subsp. oxycedrus leaves and to identify active compounds through bioactivity guided isolation technique. MATERIALS AND METHODS: Ethanol and water extracts of Juniperus oxycedrus subsp. oxycedrus (Joso), leaves on oral administration were studied using in vivo models in normal, glucose-hyperglycemic and streptozotocin-induced diabetic rats. Through in vivo bioactivity-guided fractionation processes, a nonpolar fraction was separated from the n-hexane subextract by silica gel column chromatography as the main active fraction. Subfractions of this fraction was found to possess antidiabetic activity and their chemical composition was investigated by GC-FID and GC-MS, simultaneously. RESULTS: This is the first report on the antidiabetic constituents of Joso leaves. Fatty acids, such as palmitic, linoleic and linolenic acid were found as the major compounds in subfractions. CONCLUSION: Results indicated that Joso leaf extract and its active constituents might be beneficial for diabetes mellitus.

Identification of hypoglycaemic compounds from berries of Juniperus oxycedrus subsp. oxycedrus through bioactivity guided isolation technique.

ETHNOPHARMACOLOGICAL RELEVANCE: Decoction of Juniperus oxycedrus subsp. oxycedrus L. (Cupressaceae) berries is used internally as tea and pounded fruits are consumed to lower blood glucose levels in Turkey. AIM OF THE STUDY: To evaluate hypoglycaemic and antidiabetic activity of J. oxycedrus subsp. oxycedrus berries and to identify active compounds through bioactivity guided isolation technique. MATERIAL AND METHODS: Hypoglycaemic effect of J. oxycedrus subsp. oxycedrus (Joso) berry extracts on oral administration was studied using in vivo models in normal, glucose-hyperglycaemic rats. Streptozotocin induced diabetic rats were used to examine antidiabetic activity of Joso extracts, subextracts, fractions, subfractions and shikimic acid (SA). RESULTS: Through in vivo bioactivity-guided fractionation processes, shikimic acid, 4-O-ß-d-glucopyranosyl ferulic acid and oleuropeic acid-8-O-ß-d-glucopyranoside were isolated from the n-butanol subextract by silica gel and reverse phase column chromatography as the main active ingredient of the active subfraction. After 8 days administration of the major compound shikimic acid, blood glucose levels (24%), malondialdehyde levels in kidney tissues (63-64%) and liver enzymes (AST, ALT, ALP) of diabetic rats were decreased. CONCLUSION: Results indicated that Joso berry extract and its active constituents might be beneficial for diabetes and its complications.

Insights into cholinesterase inhibitory and antioxidant activities of five Juniperus species.

In vitro acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory and antioxidant activities of the aqueous and ethanol extracts of the leaves, ripe fruits, and unripe fruits of Juniperus communis ssp. nana, Juniperus oxycedrus ssp. oxycedrus, Juniperus sabina, Juniperus foetidissima, and Juniperus excelsa were investigated in the present study. Cholinesterase inhibition of the extracts was screened using ELISA microplate reader. Antioxidant activity of the extracts was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenging, ferrous ion-chelating, and ferric-reducing antioxidant power (FRAP) assays. Total phenol and flavonoid contents of the extracts were determined spectrophotometrically. The extracts had low or no inhibition towards AChE, whereas the leaf aqueous extract of J. foetidissima showed the highest BChE inhibition (93.94 ± 0.01%). The leaf extracts usually exerted higher antioxidant activity. We herein describe the first study on anticholinesterase and antioxidant activity by the methods of ferrous ion-chelating, superoxide radical scavenging, and ferric-reducing antioxidant power (FRAP) assays of the mentioned Juniperus species.

Effects of Juniperus oxycedrus ssp. oxycedrus on tissue lipid peroxidation, trace elements (Cu, Zn, Fe) and blood glucose levels in experimental diabetes.

ETHNOPHARMACOLOGICAL RELEVANCE: Juniperus oxycedrus L. (Cupressaceae) fruits and leaves are used internally and pounded fruits are eaten for diabetes in Turkey. AIM OF THE STUDY: To evaluate the interrelationships between the levels of chosen trace elements (copper, iron, and zinc) and hepatic, renal lipid peroxidation (TBARS) in streptozotocin-induced diabetic rats treated with Juniperus oxycedrus ssp. oxycedrus (J.o.s.o.) leaf and fruit extracts for 10 days. MATERIALS AND METHODS: J.o.s.o. fruit and leaf extracts were administered in STZ-induced diabetic rats, at doses of 500 and 1000 mg/kg. The blood glucose levels were measured in the 1st, 4th, 7th and 10th day of experiment. Fe, Cu, and Zn contents and lipid peroxidation levels of liver and kidney tissues were determined by atomic absorption spectrophotometry and ultraviolet spectrophotometry, respectively. RESULTS: Treatment of diabetic rats with the J.o.s.o. fruit and leaf extracts decreased the blood glucose levels and both the levels of lipid peroxidation in liver and kidney tissues. J.o.s.o. extracts have augmented Zn concentrations in liver of STZ-induced diabetic rats. CONCLUSIONS: Results indicated that J.o.s.o. fruit and leaf extracts might be beneficial for diabetes and its complications.

Hypoglycemic activity and antioxidant potential of some medicinal plants traditionally used in Turkey for diabetes.

Cydonia oblonga Mill. (Rosaceae) leaves, Helianthus tuberosus L. (Asteraceae) tubers, and Allium porrum L. (Liliaceae) bulbs are used as a folk remedy for the treatment of diabetes and they are also consumed as food in Turkey. In the present study, the antidiabetic and antioxidant activities of the ethanol extracts of these plants were studied in normal and streptozotocin-induced diabetic rats for 5 days. All extracts were administrated orally to rats at the doses of 250 and 500 mg/kg. Blood glucose level was measured according to glucose oxidase method. In order to determine antioxidant activity, thiobarbituric acid reactive substance (TBARS) and reduced glutathione (GSH) levels in liver, kidney, and heart tissues were measured by using spectrophotometric methods. Oral administration of Cydonia oblonga (500 mg/kg) and Allium porrum (500 mg/kg) extracts for 5 days in diabetic rats caused a decrease in blood glucose levels by 33.8% and 18.0%, respectively. Moreover, Allium porrum and Cydonia oblonga extracts induced significant alleviation on only heart tissue TBARS levels (44.6 and 45.7%), Helianthus tuberosus and Allium porrum extracts showed an inhibitory effect on kidney tissue TBARS levels (24.5 and 14.8%). None of the extracts restored GSH levels in kidney, liver, and heart tissues of diabetic rats.

Antibacterial, antifungal, and antiviral activities of some flavonoids.

Antibacterial and antifungal activities of six plant-derived flavonoids representing two different structural groups were evaluated against standard strains of Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis and their drug-resistant isolates, as well as fungi (Candida albicans, C. krusei) using the microdilution broth method. Herpes simplex virus Type-1 and Parainfluenza-3 virus were employed for antiviral assessment of the flavonoids using Madin-Darby bovine kidney and Vero cell lines. Ampicillin, gentamycin, ofloxacin, levofloxacin, fluconazole, ketoconazole, acyclovir, and oseltamivir were used as the control agents. All tested compounds (32-128 microg/ml) showed strong antimicrobial and antifungal activities against isolated strains of P. aeruginosa, A. baumanni, S. aureus, and C. krusei. Rutin, 5,7-dimethoxyflavanone-4'-O-beta-D-glucopyranoside and 5,7,3'-trihydroxy-flavanone-4'-O-beta-D-glucopyranoside (0.2-0.05 microg/ml) were active against PI-3, while 5,7-dimethoxyflavanone-4'-O-[2''-O-(5'''-O-trans-cinnamoyl)-beta-D-apiofuranosyl]-beta-D-glucopyranoside (0.16-0.2 microg/ml) inhibited potently HSV-1.

Sesquiterpene lactones with antinociceptive and antipyretic activity from two Centaurea species.

ETHNOPHARMACOLOGICAL RELEVANCE: Several Centaurea species are used to alleviate pain and inflammatory symptoms in rheumatoid arthritis, high fever, and head ache in Turkish folk medicine. AIM OF THE STUDY: The effectiveness of extracts, fractions and subfractions from dried Centaurea solstitialis L. subsp. solstitialis (CSS) (Asteraceae) roots and aerial parts were studied on mice. MATERIALS AND METHODS: The antinociceptive and antipyretic effects of Centaurea solstitialis L. subsp. solstitialis have been investigated by using p-benzoquinone-induced writhing reflex for antinociceptive activity and Freund's Complete Adjuvant-induced pyrexia model for antipyretic activity assessment in mice. RESULTS: The ethanolic extract from the aerial parts of the plant was shown to possess significant antinociceptive (p < 0.01) and antipyretic activities (p < 0.01). The extract was then submitted to subsequent solvent extractions and chromatographic processes. Through bioassay-guided fractionation and isolation procedures two sesquiterpene lactones, solstitialin A and acetyl solstitialin, were isolated and defined as the active components of CSS. On the other hand, a comparative study was conducted on another species, Centaurea depressa Bieb., which has no similar folkloric utilization. Following the same fractionation chart same compounds were defined as the active ingredients. CONCLUSION: Results of the present study proved that aerial part of CSS possesses antinociceptive and antipyretic activities supporting the folkloric assertion in Turkish folk medicine. However, these effects seem not limited to CSS, some other Centaurea species, in fact, having no folkloric use might be equally active.

Hepatoprotective effect of Vitis vinifera L. leaves on carbon tetrachloride-induced acute liver damage in rats.

The hepatoprotective effect of ethanolic extract and its four different fractions (CHCl(3), EtOAc, n-BuOH, and remaining water fraction) of Vitis vinifera L. leaves was investigated against carbon tetrachloride (CCl(4))-induced acute hepatotoxicity in rats. The ethanolic extract was found active at 125mg/kg dose (per os). The ethanolic extract was fractionated through successive solvent-solvent extractions and the n-BuOH fraction in 83mg/kg dose possessed remarkable antioxidant and hepatoprotective activities. Liver damage was assessed by using biochemical parameters (plasma and liver tissue MDA [malondialdehyde], transaminase enzyme levels in plasma [AST-aspartate transaminase, ALT-alanine transferase] and liver GSH [glutathione] levels). Additionally, the pathological changes in liver were evaluated by histopathological studies. Legalon 70 Protect was used as standard natural originated drug.

In-vivo assessment of antidiabetic and antioxidant activities of grapevine leaves (Vitis vinifera) in diabetic rats.

The acute and the subacute (15 days) hypoglycaemic and antihyperglycaemic effect of the two different doses (250, 500 mg/kg) of the aqueous extract from the leaves of Vitis vinifera L. were evaluated in this study. The aqueous extract was further fractionated through successive solvent extractions and the acute effect of different doses of its subfractions, 25 mg/kg for ethylacetate fraction, 80 mg/kg for n-butanol fraction and 375 mg/kg for remaining aqueous fraction were investigated using normal, glucose-hyperglycaemic and streptozotocin-induced diabetic rats. Blood glucose levels were measured according to the glucose oxidase method. Tolbutamide was used as a reference drug at a dose of 100 mg/kg. The antioxidant activity of the test samples was studied in the liver, kidney and heart tissues of diabetic rats by measuring malondialdehyde (MDA) and glutathion (GSH) levels. All results were compared to the diabetic control groups. The results showed that EtOAc Fr. was rich in polyphenolics and possessed a significant antihyperglycaemic and antioxidant activity equipotent with the reference hypoglycaemic agent (tolbutamide), when evaluated in diabetic rats.

Anti-inflammatory and antinociceptive activity of flavonoids isolated from Viscum album ssp. album.

Viscum album L. has been used in the indigenous systems of medicine for treatment of headache and some inflammatory diseases. In order to evaluate this information, antinociceptive and anti-inflammatory activities of the five flavonoids (5,7-dimethoxy naringenin or 4',6'-dimethoxy chalcononaringenin) derivatives, isolated from the ethyl acetate fraction of the extract from V. album ssp. album, were investigated, namely 5,7-dimethoxy-flavanone-4'-O-beta-D-glucopyranoside (1), 2'-hydroxy-4',6'-dimethoxy-chalcone-4-O-beta-D-glucopyranoside (2), 5,7-dimethoxy-flavanone-4'-O-[2"-O-(5"'-O-trans-cinnamoyl)-beta-D-apiofuranosyl]-beta-D-glucopyranoside (3), 2'-hydroxy-4',6'-dimethoxy-chalcone-4-O-[2"-O-(5"'-O-trans-cinnamoyl)-beta-Dapiofuranosyl]-beta-D-glucopyranoside (4), 5,7-dimethoxy-flavanone-4'-O-[beta-D-apiofuranosyl-(1 --> 2)]-beta-D-glucopyranoside (5). For the antinociceptive activity assessment the p-benzoquinone-induced writhing test and for the anti-inflammatory activity the carrageenan-induced hind paw edema model in mice were used. The ethyl acetate fraction in a dose of 250 mg/kg as well as compounds 2 and 5 in a 30 mg/kg dose were shown to possess remarkable antinociceptive and anti-inflammatory activities per os without inducing any apparent acute toxicity as well as gastric damage.

Evaluation of the hypoglycemic effect and antioxidant activity of three Viscum album subspecies (European mistletoe) in streptozotocin-diabetic rats.

The acute hypoglycaemic effect of water and ethanolic extracts of three Viscum album subspecies, ssp. album, ssp. austriacum, ssp. abietis, were investigated in normoglycaemic and streptozotozocin-induced diabetic rats. Blood glucose level was measured according to glucose oxidase method. The antioxidant activity of these extracts was also studied in the liver, kidney and heart tissues of streptozotocin-induced diabetic rats after subacute administration. In order to determine antioxidant activity of extracts, tissue MDA and GSH levels were measured by using spectrophotometric methods. All results were compared with the diabetic control groups. The findings obtained in the experiments demonstrated that European mistletoe (Viscum album L.) subspecies possess potent antihyperglycaemic and antioxidant activity depending on host plant.

Evaluation of hepatoprotective effect of Gentiana olivieri herbs on subacute administration and isolation of active principle.

Hepatoprotective effect of Gentiana olivieri Griseb. (Gentianaceae) flowering herbs on subacute administration were studied using in vivo models in rats. For the activity assessment on carbon tetrachloride-induced hepatic damage following biochemical parameters were evaluated; plasma and hepatic tissue malondialdehyde formation, and liver tissue glutathione level, as well as plasma transaminase enzyme levels (aspartate transferase and alanine transferase). Results of biochemical tests were also confirmed by histopathological examination. Through bioassay-guided fractionation procedures isoorientin, a known C-glycosylflavone, was isolated from the ethyl acetate fraction as the active antihepatotoxic constituent by silica gel column chromatography. Isoorientin exhibited significant hepatoprotective effect at 15 mg/kg b.w. dose.