Mert Ilhan

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 β-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.

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. Dunnetts test was used to determine the significance of differences between groups. In order to compare two groups Students 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.

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.

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 plants 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.

Discovery of new antidepressant agents: In vivo study on Anthemis wiedemanniana Fisch. & Mey

ETHNOPHARMACOLOGICAL RELEVANCE Many of the currently available antidepressants have serious adverse effects and are also expensive. Traditional medicines are major sources of original drugs, and their role in effective treatment is remarkable. In Turkish folk medicine, decoctions and infusions are prepared from the flowers of Anthemis wiedemanniana Fisch. & Mey. They exert antispasmodic and sedative effects and are also used for treating urinary inflammations. The present study was designed to evaluate the antidepressant activity of A. wiedemanniana, which is used against central nervous system disorders in Turkish folk medicine. MATERIALS AND METHODS n-Hexane, ethyl acetate, and methanol (MeOH) extracts were prepared from the flowers of the plant. The antidepressant potentials of these extracts were evaluated in mouse models using the forced swimming test, tail suspension test, and antagonism of tetrabenazine-induced ptosis, hypothermia, and suppression of locomotor activity. The results were compared with those in control and reference groups, and active constituents of the plant were determined. The MeOH extract of A. wiedemanniana was subjected to various chromatographic separation techniques, leading to the isolation and identification of the active component(s). RESULTS After confirmation of the antidepressant activity, the MeOH extract was subjected to successive solvent partitioning using solvents of increasing polarity, yielding four subextracts. Each subextract was tested on the same biological activity models. Fraction B was found to have the highest activity and subjected to further chromatographic separation. Isolated germacronolide-type sesquiterpene lactones were elucidated as tatridin A (1) and tanachin (1-epi-tatridin B) (2), which were responsible for the antidepressant activity of the flowers. CONCLUSIONS This study explored the antidepressant potential of A. wiedemanniana. Using bioassay-guided fractionation and isolation techniques, tatridin A and tanachin (1-epi-tatridin B) were determined as the main active components of the flowers. Further antidepressant mechanistic studies should be conducted for exploring the activity of these compounds against depression. This study can be an important step in the discovery of newer antidepressants.