Effects of Naphthoquinones from the Roots of Onosma Armeniacum Klokov on Wound Healing.

In Turkish folk medicine, the roots of Onosma armeniacum Klokov are used to heal wounds, burns, hemorrhoids, hoarseness, dyspnea, stomach ulcers, and abdominal aches. The objective was to evaluate the plant's ethnopharmacological applications using in vivo pharmacological experimental models. In vivo linear incision and circular excision the wound models were used to assess the wound healing activity along with histopathological investigation. The active component(s) were isolated and identified after being exposed to several chromatographic separation procedures on the primary extract. The n-hexane-dichloromethane mixture extract was subjected to chromatographic separation after the wound-healing activity was confirmed. Deoxyshikonin (1), ß,ß-dimethylacrylshikonin (2), α-methyl-n-butylshikonin (3), isovalerylshikonin (4), acetylshikonin (5), ß-hydroxyisovalerylshikonin (6), and 5,8-O-dimethylacetylshikonin (7) were identified as the structures of the isolated compounds. The efficacy of O. armeniacum to heal wounds was investigated in this study. Shikonin derivatives were identified as the primary active components of the roots by bioassay-guided fractionation and isolation procedures.

Protective effect of vitamin D on imidacloprid-induced testicular injury in rats.

Introduction: The degenerative effects of imidacloprid via oxidative stress are known. Irisin is a recently discovered peptide with energy regulator and antioxidant effects. In addition, the antioxidant potential of Vitamin D has been reported in previous studies. The current study was performed to investigate the effect of Vitamin D on testis morphology and irisin immunoreactivity in imidacloprid-treated rats. Material and methods: Thirty-two Wistar albino male rats were divided into groups: control (n = 6), corn oil (n = 6), Vitamin D (n = 6), imidacloprid (n = 7) and imidacloprid + Vitamin D (n = 7). Testis tissues were used to evaluate the histopathological, biochemical and immunohistochemical changes. Oxidative state in testis tissue was determined with total antioxidant and oxidant status markers, total antioxidant status (TAS) and total oxidant status (TOS) respectively. Results: In microscopic examination, degenerative changes in the seminiferous tubule epithelium, interstitial edema and increased irisin immunoreactivity were observed in animals given imidacloprid. Also increased TOS and decreased TAS levels were measured in these animals. It was observed that Vitamin D improved the testicular damage histopathologically when compared to the imidacloprid group. However, increase in TAS levels and decrease in both TOS levels and irisin immunoreactivity were found insignificant in animals given Vitamin D. Conclusions: In the present study it was observed that Vitamin D ameliorated testis injury caused by imidacloprid. Furthermore, imidacloprid was found to increase the immunoreactivity of irisin. In the light of our findings, we conclude that the use of Vitamin D could be beneficial against testicular damage caused by imidacloprid.

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.

Evaluation of the chemopreventive effects of Ankaferd Bloodstopper in 7,12-dimethylbenz[a]anthracene-induced oral epithelial dysplasia.

OBJECTIVES: Ankaferd BloodStopper® (ABS) is an herbal extract which has been used historically as a hemostatic agent in traditional Turkish medicine. ABS comprises of standardized mixture of herbs Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum, and Urtica dioica. In addition to its hemostatic effects, the herb ABS contains some other biological effects including antioxidant and antitumoral properties. The aim of this study is to investigate the chemopreventive effects of ABS in 7,12-dimethylbenz[a]anthracene (DMBA)-induced oral epithelial dysplasia. MATERIALS AND METHODS: A total of 40 Sprague Dawley rats were randomly divided into four groups. Group 1 animals received DMBA alone, and group 2 animals received both DMBA and Ankaferd. Group 3 animals received ABS alone while group 4 animals served as control group and received only liquid paraffine. All animals were sacrificed, and tissue samples were analyzed histologically at the end of the experimental period (14 weeks). RESULTS: Histological studies have shown that the buccal pouches of animals treated with DMBA alone revealed severe dysplasia while only mild or no dysplasia were noticed in DMBA + ABS group. Ankaferd were administered to animals and control group showed no dysplasia or other oral lesions. CONCLUSION: The results suggest that Ankaferd Bloodstopper® has chemopreventive effect against DMBA-induced oral epithelial dysplasia. CLINICAL RELEVANCE: Ankaferd Bloodstopper® could be used as a supportive treatment option of cancer in oral and maxillofacial surgery since it possesses chemopreventive effect.

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.

Wound healing and anti-inflammatory activity of some Ononis taxons.

Ononis species are used for their laxative, diuretic, analgesic, anti-inflammatory, antiviral, cytotoxic and antifungal effects as well as against skin diseases for wound healing activity. In the light of this information n-hexane, ethylacetate and methanol extracts prepared from Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj., Ononis variegata L., Ononis viscosa L. subsp. brevifolia (DC) Nym. and Ononis natrix L. subsp. natrix L. were tested for their wound healing, anti-inflammatory and antioxidant activities. Linear incision and circular excision wound models and hydroxypyroline estimation assay were used for the wound healing activity. For the assessment of chronic inflammation FCA-induced arthritis and for acute inflammation carrageenan-induced hind paw edema, TPA-induced ear edema and acetic acid-induced increase in capillary permeability tests were conducted. 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) scavenging activity assay, reducing power assay and hydroxyl radical (OH-) scavenging assay were used for determining antioxidant activities of the extracts. Results showed that O. spinosa subsp. leiosperma roots ethyl acetate extract exhibited remarkable wound healing activity with the 42.6% tensile strength value on the linear incision wound model and 60.1% reduction of the wound area at the day 12 on the circular excision wound model. Hydroxyproline content of the tissue treated by O. spinosa subsp. leiosperma roots ethyl acetate extract was found to be 41.3µg/mg. Acetic acid induced increase in capillary permeability test results revealed that O. spinosa subsp. leiosperma roots ethyl acetate extract and O. spinosa subsp. leiosperma roots methanol extract inhibited inflammation by 40.4% and 35.4% values respectively. O. spinosa subsp. leiosperma roots ethyl acetate extract showed 21.2-27.2% inhibition in carrageenan-induced hind paw edema test while did not posses activity on TPA-induced ear edema and FCA-induced arthritis models.

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 and biological studies on Alnus glutinosa subsp. glutinosa, A. orientalis var. orientalis and A. orientalis var. pubescens leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Alnus species have been used for the treatment of rheumatism, hemorrhoids and for wound healing in folk medicine. MATERIALS AND METHODS: Aqueous and methanol extracts of A. glutinosa (L.) Gaertner subsp. glutinosa, A. orientalis Decne. var. orientalis and A. orientalis var. pubescens Dippel leaves were evaluated for their wound healing, anti-inflammatory and antioxidant activities. In vivo wound models of linear incision and circular excision were performed. "Inhibition of acetic acid-induced capillary permeability", "carrageenan-induced hind paw edema" and T"PA-induced ear edema" assays were applied to determine the anti-inflammatory effects. For the antioxidant activity assessment, DPPH and ABTS radicals scavenging effect, reducing power and denaturation of nonspecific hydroxyl radical-targeted 2-deoxyribose were used. In vitro inhibitory effects on enzymes hyaluronidase, collagenase and elastase were evaluated. The methanol extract of the leaves of A. glutinosa subsp. glutinosa (MB), the most potent extract, was fractionated by bioassay-guided fractionation technique. The structure of the isolated compound was determined as shikimic acid by using NMR and IR analyses. RESULTS: MB increased the wound tension by 42.79% value and provided a contraction by 51.58%. Wound tension, contraction capacity and tissue hydroxyproline levels were increased with the application of the fraction EtOAc: MeOH (Fr. D), subfraction D27-38 and shikimic acid. In the "inhibition of acetic acid-induced capillary permeability" assay, MB, Fr. D, subfraction D27-38 and shikimic acid inhibited the permeability with significant inhibition values of 30.22%, 32.46%, 38.24% and 27.19%, respectively. In carrageenan-induced hind paw edema model, MB displayed 29.1% inhibition. Likewise, Fr. D, subfraction D27-38, and shikimic acid were found to exhibit remarkable anti-inflammatory and antioxidant effects. Shikimic acid exhibited significant inhibitory effect (38.24%) on the hyaluronidase enzyme. CONCLUSIONS: This is the first and unique study that investigates wound healing, anti-inflammatory and antioxidant effects of some Alnus taxons growing in Turkey. According to the results, shikimic acid was found to be the major compound responsible from the activity.

Investigation of the effect of crocin pretreatment on renal injury induced by infrarenal aortic occlusion.

OBJECTIVE: Ischemia and reperfusion (IR) injury is an important complication of abdominal aortic surgery, and it mainly affects the lower extremities and remote organs. In the present study, we aimed to investigate the possible protective effect of crocin in IR-mediated kidney damage. MATERIALS AND METHODS: A total of 24 adult male Wistar-Albino rats were equally and randomly separated into three groups as follows: sham laparotomy, IR, and IR + crocin. Infrarenal aortic occlusion and reperfusion was applied for 1 and 2 h, respectively. Tissue samples were removed and collected. Biochemical and histopathologic analyses were performed. RESULTS: Urea, blood urea nitrogen, creatinine, renal tissue tumor necrosis factor α, interleukin (IL)-6, IL-18, interferon gamma, IL-1ß, total oxidant status, and oxidative stress index levels were significantly higher in IR group, when compared with other groups. These improvements were also demonstrated with some parameters including total score of histopathologic damage, Tunel, Bax, and Caspase-3 expression levels, and these parameters were prominently higher in the IR group, when compared with the other groups. Nevertheless, Bcl2 expression degree was prominently lower in the IR group than those in the other two groups. CONCLUSIONS: Data established from the present study suggest that crocin can preclude renal damage in infrarenal aortic occlusion models.

Activity of Corylus avellana seed oil in letrozole-induced polycystic ovary syndrome model in rats

Abstract The aim of the present study was to assess the activity of the hazelnut oil in the treatment of polycystic ovary syndrome in rats. Serum follicle-stimulating hormone, luteinizing hormone, estradiol, progesterone, testosterone, serum lipid parameters, leptin and glucose levels were evaluated. Moreover, antioxidant activity was tested using superoxide dismutase, malondialdehyde, catalase, glutathione peroxidase levels. The phytosterol content of the oil was determined by HPLC. The plasma high density lipoprotein-cholesterol level was found to be significantly high and leptin and glucose concentrations were found to be significantly low in the treatment group. According to the phytochemical analysis, the main components of the oil were detected as α-tocopherol, γ-tocopherol, squalene, β-sitosterol, campesterol and stigmasterol. Corylus avellana oil was found to be effective in the treatment of polycystic ovary syndrome via regulating gonadotropins, steroids and serum lipid parameters and possesses antioxidant activity.

A mixture of St. John's wort and sea buckthorn oils regresses endometriotic implants and affects the levels of inflammatory mediators in peritoneal fluid of the rat: A surgically induced endometriosis model.

OBJECTIVE: Sea buckthorn (Hippophae rhamnoides L.) and St. John's wort (Hypericum perforatum L.) are used as an emmenagog and for the treatment of other gynecological disorders including uterus inflammation and endometriosis. The aim of the present study is to investigate the potential of a mixture of sea buckthorn and St. John's wort oils (HrHp oil) in the treatment of endometriosis. MATERIALS AND METHODS: The activity was assessed in surgically induced endometriosis in rats. A 15-mm piece of endometrium was sutured into the abdominal wall. Twenty-eight days later, a second laparotomy was performed to calculate the endometrial foci areas and to score intra-abdominal adhesions. The rats were treated with either vehicle, HrHp oil formulation, or the reference (buserelin acetate). At the end of the experiment all rats were sacrificed and endometriotic foci areas and intra-abdominal adhesions were re-evaluated. The tissue sections were analyzed histopathologically. Peritoneal fluids of the experimental animals were collected in order to detect the levels of tumor necrosis factor-α, vascular endothelial growth factor, and interleukin-6, which might be involved in the etiology of endometriosis. RESULTS: In the HrHp oil-treated group, the volumes of endometriotic implants were found to be significantly decreased (from 50.8 mm3 to 18.6 mm3, p<0.001) without any adhesion (0.0±0.0, p<0.001) when compared to the control group (3.1±0.9). The levels of tumor necrosis factor-α decreased from 7.02±1.33 pg/mL to 4.78±1.02 pg/mL (p<0.01); vascular endothelial growth factor from 17.39±8.52 pg/mL to 9.67±5.04 pg/mL (p<0.01); and interleukin-6 from 50.95±22.84 pg/mL to 29.11±7.45 pg/mL (p<0.01), respectively, after HrHp oil treatment. CONCLUSION: HrHp oil may be a promising alternative for the treatment of endometriosis.

Neurodegenerative changes and apoptosis induced by intrauterine and extrauterine exposure of radiofrequency radiation.

Adverse health effects of radiofrequency radiation (RFR) on the ongoing developmental stages of children from conception to childhood are scientifically anticipated subject. This study was performed to identify the effects of global system for mobile communications (GSM) modulated mobile phone like RFR in 1800MHz frequency on oxidative DNA damage and lipid peroxidation beside the apoptotic cell formation, using histopathological and immunohistochemical methods in the brain tissue of 1-month-old male and female New Zealand White rabbits that were exposed to these fields at their mother's womb and after the birth. Oxidative DNA damage and lipid peroxidation levels were investigated by measuring the 8-hydroxy-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels, respectively. Histopathological changes were observed using by hematoxylin and eosin (HE) staining. Apoptotic cells were detected in the examined organs by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining. For both male and female infants; 8-OHdG levels increased in the group exposed to RFR in both intrauterine and extrauterine periods compared to the infants that were never exposed to RFR and the ones were exposed when they reached one month of age (p<0.05). MDA results were different for male and female rabbits. There was no difference between all female infant groups (p>0.05), while only intrauterine exposure significantly causes MDA level increase for the male infants. HE staining revealed mild lessions in neuronal necrobiosis in brain tissues of female rabbits that had only intaruterine exposure and male rabbits had only extrauterine exposure. Gliosis were mildly positive in brain tissues of rabbits that are exposed only intrauterine period, also the group exposed both intrauterine and extrauterine periods. However, there was no apoptotic change detected by TUNEL staining in the brain tissues of all groups.

Topical application of olive oil macerate of Momordica charantia L. promotes healing of excisional and incisional wounds in rat buccal mucosa.

OBJECTIVE: In Turkish folk medicine Momordica charantia L. is used for wound healing. The aim of the present study is to investigate this folkloric knowledge and confirm the plant's potential effect on buccal mucosa wound in the rat. DESIGN: Wound healing activity of olive oil macerate of Momordica charantia L. was investigated in linear incision and circular excision wound models created in the buccal mucosa of the rat. The tissues were histopathologically evaluated, moreover, hydroxyproline contents of the tissues were determined. The anti-inflammatory activity was also assessed by using Whittle method with some modifications. RESULTS: Olive oil macerate of M. charantia showed significant wound healing activity both in incision (45.1%) and excision (89.8%) wound models and demonstrated anti-inflammatory activity with the inhibition value of 31.3% at the dose of 100mg/kg. CONCLUSION: The experimental data revealed that M. charantia showed significant wound healing and anti-inflammatory effect.

The effects of amlodipine and platelet rich plasma on bone healing in rats.

AIM: The aim of this study was to evaluate the effects of calcium channel blocker (CCB) amlodipine (AML), platelet rich plasma (PRP), and a mixture of both materials on bone healing. MATERIALS AND METHODS: Fifty-six male Wistar rats were randomly divided into four groups: group A, tibia defect model with no treatment; group B, tibia defect model treated with AML, 0.04 mg daily by oral gavage; group C, tibia defect model treated with local PRP; group D, tibia defect model treated with local PRP and AML, 0.04 mg daily by oral gavage. RESULTS: At day 21, bone healing was significantly better in groups C and D compared to group A (P<0.05), but comparisons showed no statistically significant difference in group B (P>0.05). At day 30, groups B and C showed no statistically significant difference (P>0.05) compared to group A, but bone healing in group D was significantly better than in group A (P<0.05). Statistically, AML did not affect alkaline phosphatase (ALP) activity at 21 and 30 days (P>0.05), but PRP and AML + PRP increased ALP activity statistically (P<0.05). CONCLUSION: It can be concluded that AML had neither a positive nor a negative effect on bone healing, but when used in combination with PRP, it may be beneficial.

Thuja occidentalis L. and its active compound, α-thujone: Promising effects in the treatment of polycystic ovary syndrome without inducing osteoporosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Thuja occidentalis L. (Cupressaceae) has been used in folk medicine for the treatment of rheumatism, amenorrhea, cystitis, and uterine carcinomas, and as an abortifacient and contraceptive. AIM OF THE STUDY: The present study aimed to determine whether T. occidentalis oil and α-thujone could be beneficial in the treatment of polycystic ovary syndrome (PCOS). MATERIALS AND METHODS: T. occidentalis oil and α-thujone were administered to rats with letrozole-induced PCOS for 21 days. At the end of 21 days, the rats were sacrificed and blood samples were taken by cardiac puncture. The levels of serum gonadotropins, steroids, blood lipid, leptin, and glucose and the values of antioxidant parameters were measured. RESULTS: The results demonstrated that estradiol and progesterone levels significantly increased, while luteinizing hormone (LH) and testosterone levels decreased in the T. occidentalis- and α-thujone-administered groups. The plasma low-density lipoprotein-cholesterol (LDL-C), leptin, and glucose concentrations were also significantly decreased in the T. occidentalis and α-thujone groups when compared to the control group. Histopathological findings demonstrated that the T. occidentalis and α-thujone groups displayed good healing. According to the phytochemical analyses, 25 compounds were identified in the T. occidentalis oil. The main constituents of the oil were the monoterpene ketones α- and ß-thujone, fenchone, and sabinene, as well as the diterpenes beyerene and rimuene. CONCLUSION: T. occidentalis essential oil and its active component, α-thujone, can be used for the treatment of PCOS without inducing osteoporosis.

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.

Prevention of cyclophosphamide-induced hemorrhagic cystitis by resveratrol: a comparative experimental study with mesna.

PURPOSE: Hemorrhagic cystitis (HC) is the most common urotoxic side effect of cyclophosphamide (CP). The aim of this study was to compare the classical efficacy of mesna (2-mercaptoethane sulfonate sodium) with three different doses of resveratrol (RES) on cyclophosphamide-induced HC in rats. METHODS: Forty-six male Sprague-Dawley rats were divided into six groups. Group 1 served as a negative control (sham). Five groups received a single dose of cyclophosphamide (150 mg/kg) intraperitoneally at the same time. Groups 2, 3, 4, 5, and 6 received only CP, CP + 20 mg/kg RES, CP + 40 mg/kg RES, CP + 80 mg/kg RES, and CP + classical protocol of three doses of mesna (30 mg/kg three times), respectively. Antioxidants, cytokines, and malondialdehyde levels were measured in all groups. In addition, histopathological alterations in tissues were examined. RESULTS: CP administration induced severe HC with marked edema, hemorrhage, and inflammation in group 2. RES 20 mg/kg showed meaningful protection against bladder damage compared to the control group. It was seen that RES 40 mg/kg gave weaker protection but RES 80 mg/kg was not found to be effective. CONCLUSION: In conclusion, marked bladder protection was found in 20 and 40 mg/kg RES applications compared to the control group, but this protection was weaker than with mesna.

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.

Experimental endometriosis remission in rats treated with Achillea biebersteinii Afan.: histopathological evaluation and determination of cytokine levels.

OBJECTIVE: The aim of the present study is to assess the beneficial effects of Achillea biebersteinii Afan. in the treatment of endometriosis in order to find scientific evidence for the folkloric use of this plant. STUDY DESIGN: Experimental endometriosis was induced in six-week-old female, nonpregnant, Sprague Dawley rats by suturing a 15mm piece of endometrium from uterine cornu into abdominal wall. After twenty-eight days, a second laparotomy was performed: the endometrial foci areas were measured and intra-abdominal adhesions were scored, and the abdomen was closed. Different groups then received n-hexane, ethyl acetate (EtOAc) and methanol (MeOH) extracts prepared from the aerial parts of A. biebersteinii, and a control group received inert material, administered per os once a day throughout the experiment. At the end of the treatment procedure all rats were sacrified and endometriotic foci areas and intra-abdominal adhesions were again evaluated and compared with the previous findings. The tissues were also histopathologically investigated. Moreover, peritoneal fluid was collected to detect tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) levels. Dunnett's test was used to determine the significance of differences between groups. In order to compare two groups Student's t test was used. RESULTS: Post-treatment volumes of endometrial foci were found to be significantly decreased, and no adhesion was detected, in the EtOAc extract treated group. The levels of TNF-α, VEGF and IL-6 also fell after the treatment with EtOAc extract. The therapeutic effect of the EtOAc extract of A. biebersteinii could be attributed to the flavonoid aglycones found in the extract. CONCLUSION: The EtOAc extract of A. biebersteinii appears to be a promising alternative for the treatment of endometriosis.

Exploration of the wound healing potential of Helichrysum graveolens (Bieb.) Sweet: isolation of apigenin as an active component.

ETHNOPHARMACOLOGICAL RELEVANCE: In Turkish traditional medicine, the flowers of Helichrysum graveolens (Bieb.) Sweet (Asteraceae) have been used for the treatment of jaundice, for wound-healing and as a diuretic. AIM OF THE STUDY: In order to find scientific evidence for the traditional utilization of this plant in wound-healing, the effect of the plant extract was investigated by using in vivo and in vitro experimental models. Then through bioassay-guided fractionation procedures active wound-healing component(s) was isolated and its possible role in the wound-healing process was also determined. MATERIAL AND METHODS: The linear incision and the circular excision wound models were applied in order to evaluate in vivo wound-healing potential of Helichrysum graveolens. Anti-inflammatory and antioxidant activities, which are known to involve in wound-healing process, were also assessed by the Whittle method and the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging assay, respectively. The total phenolic content of the crude extract and solvent fractions was estimated to find correlation between the phenolic content and the antioxidant activity. Combined application of the chromatographic separation techniques on sephadex and silica gel columns, and bioassay techniques have yielded the active wound-healing principle of Helichrysum graveolens. Moreover, in vitro inhibitory effect of active principle on hyaluronidase, collagenase and elastase enzymes were investigated to explore the activity pathways. RESULTS: The 85% methanol (MeOH) extract of Helichrysum graveolens flowers displayed significant wound-healing, anti-inflammatory and antioxidant activities. Then the crude extract was partitioned by successive solvent extractions, in increasing polarity, to give five solvent fractions. Among the solvent fractions, the ethyl acetate (EtOAc) fraction exerted the highest activity. The EtOAc fraction was further subjected to chromatographic separations to yield active constituent and its structure was elucidated to be apigenin by spectrometric methods. Further in vivo and in vitro assays revealed that apigenin was one of the components responsible for the wound-healing effect of the plant remedy and also found to possess significant anti-inflammatory, antioxidant, anti-hyaluronidase and anti-collagenase activities. CONCLUSION: Present study supported the traditional use of Helichrysum graveolens flowers for wound-healing and through bioassay-guided fractionation procedures from the crude extract apigenin was isolated as one of the active components.