Evaluation of the Antidiabetic Activity of Alchemilla persica Rothm. in Mice with Diabetes Induced by Alloxan.

OBJECTIVES: Alchemilla species are used in Turkish folk medicine for the treatment of many diseases together with diabetes. Alchemilla persica, belonging to this genus, is widely distributed in Eastern Anatolia as well as in Caucasia, northern and northeastern Iran, and northern Iraq. MATERIALS AND METHODS: Methanolic-water extracts of the aerial parts and roots of A. persica were evaluated for their hypoglycemic activities in mice with alloxan-induced diabetes in order to verify its usage in folk medicine. RESULTS: None of the tested extracts exhibited a significant lowering effect on blood glucose levels. However, the aerial parts notably increased blood glucose levels at doses of 100 mg/kg and 200 mg/kg. CONCLUSION: A. persica usage as an antidiabetic is not confirmed in the present study.

Antidiabetic activity evaluation of Onobrychis species on alloxan-induced diabetic mice

In the current study, four Onobrychis species, O. albiflora Hub.-Mor., O. argyrea Boiss. subsp. argyrea Boiss., O. galegifolia Boiss., and O. tournefortii (Willd.) Desv. were collected from Anatolia to be evaluated for their antidiabetic activities. Methanol water extracts of the aerial parts were used for experiments. An alloxan-induced diabetic mice test model was used. Phytochemical analysis of the tested extracts was investigated using the HPLC method. The highest activity was observed with treatment of O. albiflora aerial part extract. Significant decrements were detected in the blood glucose levels as follows: 180.83±47.48 and 252.83±50.47mg/dL at 100 mg/kg and 200 mg/kg doses of O. albiflora, respectively, when compared to the isotonic saline solution control group, eliciting a blood glucose level of 494.20±27.32. Among the tested standard compounds, rutin and isoquercetin were detected in the examined species. The highest amount of rutin (1.1981±0.0017%) and isoquercetin (0.7318±0.0197%) were found in O. albiflora and O. argyrea subsp. argyrea, respectively. Antidiabetic activities of the tested Onobrychis species seem to indicate a possible correlation with their rutin and isoquercetin contents. Therefore, rutin and isoquercetin may be the antidiabetic compounds that contribute to the antidiabetic activity of the tested Onobrychis species.

Phytochemical Analysis of Podospermum and Scorzonera n-Hexane Extracts and the HPLC Quantitation of Triterpenes.

Previously tested n-hexane extracts of the Scorzonera latifolia showed promising bioactivity in vivo. Because triterpenes could account for this activity, n-hexane extracts were analyzed by HPLC to identify and quantify the triterpenes as the most abundant constituents. Other Scorzonera and Podospermum species, potentially containing triterpenic aglycones, were included in the study. An HPLC method for simultaneous determination of triterpene aglycones was therefore developed for analysis of Podospermum and Scorzonera species. n-Hexane extracts of root and aerial parts of S. latifolia, ten other Scorzonera species and two Podospermum species were studied to compare the content of triterpenes. HPLC was used for the qualitative and quantitative analysis of α-amyrin, lupeol, lupeol acetate, taraxasteryl acetate, 3-ß-hydroxy-fern-7-en-6-one acetate, urs-12-en-11-one-3-acetyl, 3-ß-hydroxy-fern-8-en-7-one acetate, and olean-12-en-11-one-3-acetyl. Limits of detection and quantification were determined for each compound. HPLC fingerprinting of n-hexane extracts of Podospermum and Scorzonera species revealed relatively large amounts of triterpenes in a majority of investigated taxa. Lupeol, lupeol acetate, and taraxasteryl acetate were found in a majority of the species, except S. acuminata. The presence of α-amyrin, 3ß-hydroxy-fern-7-en-6-one-acetate, urs-12-en-11-one-3-acetyl, 3ß-hydroxy-fern-8-en-7-one-acetate, and olean-12-en-11-one-3-acetyl was detected in varying amounts. The triterpene content could correlate with the analgesic and anti-inflammatory activity of Scorzonera, which was previously observed and Scorzonera species that have been determined to contain triterpenes in large amounts and have not yet been tested for their analgesic activity should be tested for their potential analgesic and anti-inflammatory potential. The presented HPLC method can be used for analysis of triterpene aglycones, for example dedicated to chemosystematic studies of the Scorzonerinae.

In vivo wound-healing activity of Euphorbia characias subsp. wulfenii: Isolation and quantification of quercetin glycosides as bioactive compounds.

ETHNOPHARMACOLOGICAL RELEVANCE: The latex and the aerial parts of Euphorbia characias L. (Euphorbiaceae) have been used as medicinal plant to treat wounds and warts in traditional medicine. AIM OF THE STUDY: The effect of the plant extract was tested in vivo and in vitro with experimental models to find scientific evidence for traditional use in wound healing. Potentially active wound-healer compounds were isolated from the active fraction using fractionation procedures under the guidance of biological assay and the possible role of the compounds in the wound healing process was also determined. MATERIAL AND METHODS: N-hexane, ethyl acetate, and methanol extracts were successively prepared from the aerial parts of E. characias subsp. wulfenii. The extracts were tested with linear incision, circular excision wound models and the hydroxyproline assay method to assess the wound-healing activity. The inhibition of the increase in capillary permeability induced by acetic acid, an acute inflammation model, was used to assay the anti-inflammatory activity. Different chromatographic separation techniques on sephadex and silica gel columns, and bioassay guided assay techniques have been used to isolate the active compounds of the plant. Moreover, hyaluronidase, collagenase and elastase enzymes inhibitory effect of active principle were investigated in vitro to find out the mechanism of action. RESULTS: The methanol (MeOH-ex) extract of the aerial parts of E. characias subsp. wulfenii showed significant wound healing activity (linear incision wound model: 43.04%; circular excision wound model 65.24%) and anti-inflammatory activity (34.74%). The methanol extract was separated into its fractions by column chromatography for isolation of efficient compounds. Biological activity of the fractions were assessed and further isolation and purification processes have been carried out in the active fraction. Isolation studies were carried out from the MeOH-ex fraction to obtain active constituents and their structures were elucidated to be quercetin-3-O-rhamnoside (quercitrin), quercetin-3-O-galactoside (hyperoside), and quercetin-3-O-arabinoside (guaijaverin). Further in vitro and in vivo assays showed that quercetin derivatives were responsible for the wound-healing activity of the plant, and also found to be significant anti-elastase and anti-collagenase activities. The amounts of three compounds, isolated from active fraction, were determined by using high performance liquid chromatography. Calibration equation was calculated with dilutions, prepared from pure substances, and assay was performed in total extract, prepared from E. characias subsp. wulfenii. It was detected that the plant had 1.22% quercitrin, 0.35% hyperoside, and 0.11% guaijaverin. The validation of the analytical method was performed by linearity, precision, limit of detection, and limit of quantification parameters. CONCLUSION: Present study supported the traditional use of the aerial parts E. characias subsp. wulfenii as wound healer and quercetin derivatives were isolated as active components from the active fraction by using bioassay-guided fractionation technique.

In vivo activity assessment of some Tanacetum species used as traditional wound healer along with identification of the phytochemical profile by a new validated HPLC method.

OBJECTIVES: Tanacetum species are traditionally used as insecticide, and externally wound healer as well as for anti-inflammatory and antihistaminic properties. The in vivo wound-healing and anti-inflammatory potential of four Tanacetum species, Tanacetum argenteum (Lam.) Willd. subsp. argenteum (TA), Tanacetum heterotomum (Bornm.) Grierson (TH), Tanacetum densum (Lab.) Schultz Bip. subsp. sivasicum (TD), and Tanacetum vulgare L. (TV) was investigated. MATERIALS AND METHODS: The chloroform (CHCl3) and methanol:water (80:20) extracts were prepared from the aerial parts of each plant. For assessment of the wound-healing activity, linear incision on rats and circular excision on mice wound models were used and histopathological analyses were conducted on the tissues treated with the test materials. For the evaluation of the anti-inflammatory activity, Whittle Method based on the inhibition of the acetic acid-induced increase in capillary permeability was used. In order to elucidate the phytochemical contents of the extracts, HPLC profiles of active fractions were screened and quantitative analysis was conducted within the scope of HPLC analysis. RESULTS: The CHCl3 extracts of TD, TA and TV were found to have significant wound healing activity (37.1%, 30.8% and 26.1% tensile strength; 88.05%, 72.93% and 44.88% contraction values, respectively) and anti-inflammatory activities (31.5% and 26.6% inhibition values for TD and TA). Parthenolide content of the CHCl3 extracts of TA, TH and TV were found 242.66±1.53, 190.16±5.62 and 177.51±3.73 µg/100 mg plant material, respectively. CONCLUSION: According to the results, the other secondary metabolites present in the aerial parts of the Tanacetum species possibly exerted synergistic effects on the observed healing of the wounds.

Evaluation of hepatoprotective and antidiabetic activity of Alchemilla mollis.

Alchemilla mollis (Buser) Rothm aerial part and root methanolic-water extracts were evaluated for their hepatoprotective activity on carbon tetrachloride induced hepatotoxicity and hypoglycemic activity on alloxan-induced diabetic mice. None of the tested extracts exhibited effects on blood glucose levels. However hepatoprotective activity results have revealed that serum ALT levels were significantly lowered by both the aerial part and root extracts at doses of 100mg/kg and 200mg/kg. Histopathological examination showed that A. mollis aerial parts and roots induced significant recovery from cellular damage; when compared to the carbon tetrachloride group, the most significant activity was observed with A. mollis aerial part extracts at a dose of 200mg/kg. There is evidence of a hepatoprotective activity of A. mollis on the phenolic content of the plant, especially in the case of flavonoids, which have potent antioxidant properties.

Viburnum opulus L.: A remedy for the treatment of endometriosis demonstrated by rat model of surgically-induced endometriosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Endometriosis is a gynecological disorder characterized by the presence of endometrial tissue outside the uterine cavity. The fruits of Viburnum opulus L. have been used to treat gynecological disorders including primary and secondary dysmenorrhea and ovarian cysts. MATERIALS AND METHODS: Air-dried and powdered fruits of V. opulus were extracted sequentially with n-hexane, ethyl acetate (EtOAc), and methanol (MeOH), respectively for four days. Endometriosis was induced by suturing 15mm piece of endometrium into abdominal wall of Sprague Dawley rats. In second laparotomy, the dimensions of endometrial implants were measured and intra-abdominal adhesions were scored. The abdomen was closed. Extracts were daily administered to the rats. At the end of the experiment, rats were sacrified and endometriotic foci areas and intra-abdominal adhesions were re-evaluated. The tissues were also histopathologically investigated. Furthermore, tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) levels of peritoneal fluid were measured. HPLC analyses were conducted on the most potent EtOAc and MeOH extracts to determine the amount of the major compound, chlorogenic acid. RESULTS: The endometriotic volumes were found to be reduced significantly in the EtOAc extract-administered group to 30.1mm3 and in the MeOH extract-administered group to 34.7mm3 as compared to the control group. No adhesion was observed in the reference and EtOAc groups. Histopathological data also supported the results. Both EtOAc and MeOH extract-administered groups displayed significant remission in the levels of TNF-α, VEGF and IL-6. When the active extracts were subjected to HPLC analysis, chlorogenic acid was found to be the major compound and the amount of this compound was calculated as 0.5112±0.0012mg and 1.7072±0.0277mg/100mg extract, repectively. CONCLUSIONS: The results of the present study indicate that the effectiveness of the fruit extract of V. opulus could be partially attributed chlorogenic acid. Other phenolic compounds could potentiate the activity due to their amount.

Phytochemical analyses and effects of Alchemilla mollis (Buser) Rothm. and Alchemilla persica Rothm. in rat endometriosis model.

PURPOSE: The aim of the present study is to evaluate the treatment potential of Alchemilla mollis (Buser) Rothm. and Alchemilla persica Rothm. in the experimentally induced endometriosis model in rats. METHODS: Endometriosis was surgically induced in rats by autotransplanting endometrial tissue to abdominal wall. Thirty-six rats were randomly divided into six groups. The groups were orally treated with the methanol:water (80:20) extracts of aerial parts and roots of A. mollis and A. persica. Buserelin acetate (20 mg) was used as the reference drug. The phytochemical contents of the most active extracts were determined by high performance liquid chromatography. RESULTS: The cystic formation was determined to be significantly decreased with the aerial part extract of A. mollis. A reduction in the endometrioma was also determined for the aerial part extract of A. persica group. However, significant reduction on the levels of cytokine were recorded for the A. mollis aerial part extract group. Therefore, the phytochemical contents of the aerial part extracts of A. mollis. and A. persica were analyzed. CONCLUSION: The results of the present study revealed that the aerial part extracts of A. mollis and A. persica could be beneficial in the treatment of endometriosis.

Preclinical Evaluation of Antiurolithiatic Activity of Viburnum opulus L. on Sodium Oxalate-Induced Urolithiasis Rat Model.

The aim of the present research is to evaluate the antiurolithiatic effect of the various extracts prepared from the fruits of Viburnum opulus L., in regard to its ethnobotanical record. To induce urolithiasis, 70 mg/kg sodium oxalate was injected to the rats which were housed individually in metabolic cages. The test materials were applied during 7 days. Biochemical (urine and serum parameters), histopathological and antioxidant (TBARs, TSH and GSH) assays were conducted. The urine samples were examined by light microscope for the determination of the calcium oxalate crystals. Lyophilized juice of V. opulus (LJVO) and lyophilized commercial juice of V. opulus (LCJVO) exerted potential antiurolithiatic activity which was attributed to its diuretic effect along with the inhibitory action on the oxalate levels and free radical production. We also determined the chlorogenic acid content of the LJVO by high-performance liquid chromatography (HPLC). Chlorogenic acid was determined by using Supelcosil LC-18 (250 × 4.6 mm, 5 µm) column and acetonitrile: water: 0.2% o-phosphoric acid as a mobile phase. The chlorogenic acid content of V. opulus was found to be 0.3227 mg/mL in fruit juice. The results obtained in this study have provided a scientific evidence for the traditional usage of V. opulus on passing kidney stones in Turkish folk medicine.