|
 
 |
| ORIGINAL ARTICLES |
|
| Year : 2022 | Volume
: 11
| Issue : 1 | Page : 79-84 |
|
The therapeutic effect of methanolic extract Bryonia dioica Jacq. in a female rat model of polycystic ovary syndrome
Reza Tahvilian1, Mohammad Mehdi Gravandi2, Tayebeh Noori1, Abdulhamid Papzan3, Nasim Jamshidi2, Amin Iranpanah1, Afsaneh Moradi1, Samira Shirooie1
1 Pharmaceutical Sciences Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran
2 Student Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran
3 Department of Agriculture Extension and Education, College of Agriculture, Razi University, Kermanshah, Iran
| Date of Submission | 21-Apr-2021 |
| Date of Acceptance | 16-Nov-2021 |
| Date of Web Publication | 29-Jun-2022 |
Correspondence Address:
Dr. Samira Shirooie
Pharmaceutical Sciences Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, 6734667149
Iran
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/jrptps.JRPTPS_48_21
Objective: Polycystic ovary syndrome (PCOS) is one of the most common endocrine diseases that affects 5%–10% of women of childbearing age. Several factors contribute to the development of PCOS such as dysfunction of the hypothalamic–pituitary axis and ovarian function, as well as increased insulin levels. The manifestations of the disorder include a wide range of symptoms, including menstrual disorders, acne, infertility, and increased body fat. Currently, the most well-known treatments for PCOS are clomiphene, metformin, letrozole, and tamoxifen. Due to their side effects, the identification of substitute drugs is essential. One of the traditional medicines, which is usually used in different parts of the world, particularly in Western Europe, is Bryonia dioica Jacq. (B. dioica). This plant is used in the treatment of disease due to its active ingredients like polyphenols. Materials and Methods: Induction PCOS in a female rat (3 weeks old) was performed through subcutaneous injection of testosterone enanthate (1 mg/100g) daily for 35 days. The effects of B. dioica (30 and 60 mg/kg) root methanolic extract on PCOS-induced was evaluated after 28-day treatment. On the last day, the serum levels of follicle-stimulating hormone (FSH), glucose, low-density lipoprotein/high-density lipoprotein (LDL/HDL), luteinizing hormone (LH), and testosterone and histological studies (hematoxylin and eosin [H&E] staining) were measured. Results: Results showed that FSH and LH levels (P < 0.05) as well as glucose (P < 0.001) in the B. dioica groups normalized significantly compared to the PCOS group. LDL levels decreased in rats and the LDL/HDL ratio decreased in all treatment groups. In histologic assay, metformin and B. dioica restricted the effects of testosterone in the ovaries of rats. Conclusion: The data indicate that methanolic extract of B. dioica recovers hormonal factors in PCOS. Keywords: Bryonia dioica Jacq., methanolic extract, polycystic ovary syndrome, rat, testosterone
How to cite this article:
Tahvilian R, Gravandi MM, Noori T, Papzan A, Jamshidi N, Iranpanah A, Moradi A, Shirooie S. The therapeutic effect of methanolic extract Bryonia dioica Jacq. in a female rat model of polycystic ovary syndrome. J Rep Pharma Sci 2022;11:79-84 |
How to cite this URL:
Tahvilian R, Gravandi MM, Noori T, Papzan A, Jamshidi N, Iranpanah A, Moradi A, Shirooie S. The therapeutic effect of methanolic extract Bryonia dioica Jacq. in a female rat model of polycystic ovary syndrome. J Rep Pharma Sci [serial online] 2022 [cited 2023 Sep 26];11:79-84. Available from: https://www.jrpsjournal.com/text.asp?2022/11/1/79/348877 |
| Introduction | |  |
Polycystic ovary syndrome (PCOS) is an endocrine disease that affects 5%–10% of women of childbearing age. This disease, which is an ovarian disorder, leads to adrenal–pituitary–hypothalamic disorder.[1],[2] In this disease, the secretion of luteinizing hormone (LH) increases compared to follicle-stimulating hormone (FSH). Increased LH stimulates ovarian follicular cells and increases androgen synthesis and secretion.[3],[4] Diagnosis of PCOS is based on criteria such as hyperandrogenism (clinical manifestations of androgen enhancement such as hair loss and obesity around menstruation), decreased or no chronic ovulation, and measurement of testosterone, prolactin, dehydroepiandrosterone (DHEA), and hemostasis levels. Common symptoms of PCOS include insulin resistance, hyperandrogenism, anovulation, and subsequent infertility.[4],[5] The most common treatments for this disease are clomiphene, metformin, letrozole, and tamoxifen. These medications have side effects, including hot flashes, arthritis, muscle aches, and psychological side effects such as irritability, mood swings, and depression. Treatment is done to alleviate symptoms, correct sexual cycles and ovulation, restore fertility, and prevent long-term complications.[6] Due to the side effects of using many chemicals, the use of medicinal plants is recommended. The active ingredients in plants have a biological equilibrium state due to their association with other substances. These substances do not accumulate in the body and do not produce side effects, which in this regard have a significant advantage over chemical drugs.[7],[8]
Bryonia dioica Jacq. (B. dioica) belongs to the Cucurbitaceae family. The Persian name of the plant is Hazar Goshan, Fashra, or Qira. Fashra is a perennial herbaceous plant with a height of about 3 m. This plant is fast growing and has fleshy and grooved stems, and it grows in a twisting and ascending manner. The male flowers are in clustered inflorescences and the female flowers are in umbrella inflorescences, and all flowers are yellow with green or white stripes. The root of the plant is thick and fleshy with gray skin and white and viscous inside. This plant is one of the native plants of our country and grows in Kermanshah province. In traditional medicine, various effects of this plant have been mentioned, including the effects of emetic, local analgesic, and laxative.[9],[10] Antidiabetic activities, antibacterial activities, anticancer activities, and antioxidant activities are among the important biological and therapeutic effects of B. dioica.[10]
There are two active ingredients in this plant called bryodine and cucurbitacine, whose antitumor effects have been proven. In terms of chemical composition, the plant’s root contains a bitter substance called glucoside called brionine. This glucoside is soluble in water and alcohol, and in addition, an amount of essential oil and resin called brioresin and a large amount of starch are found in the roots.[11],[12] In addition, B. dioica contains active compounds such as alkaloids, polyphenols, saponins, and flavonoids.[13] It has been indicated that polyphenols showed protective effects in PCOS via reducing inflammation, reactive oxygen species (ROS) generation, insulin resistance, and serum level of testosterone.[14] The other active ingredient in B. dioica is saponins that previous studies have been shown saponins reduced PCOS symptoms by reducing nerve growth factor (NGF) expression in ovaries and brain tissues.[15]
As no study was conducted on the B. dioica extract in PCOS, in this study the effect of methanolic extract to of B. dioica root on testosterone enanthate-induced PCOS rats was examined.
| Materials and Methods | | |
Preparation of Bryonia dioica extract
First, the plant was collected in spring from Kermanshah province and then identified by a botanist at Razi University and compared with herbarium specimens (voucher number: 128). The plant was thoroughly washed and placed on a wire rack in a room away from direct sunlight to dry. The dried plant was powdered with a special mill and then 100 g powder are infused in 100-mL methanol 70% and the extract was prepared by soaking. After 3 days, the extract was filtered using filter paper and dried by a rotary operator.[16] The dried extract was stored in the refrigerator until use (yield 15%).
Animals
Immature female Wistar rats (21 days old) (40–70 g) were procured from the central animal house, Kermanshah University of Medical Sciences and maintained at standard conditions. All the experimental procedures were performed according to the Ethical Committee of Kermanshah University of Medical Sciences (Protocol no. IR.KUMS.REC.1399.349).
Experimental design
Androgenized Rodent Model of PCOS is used to induce PCOS.[17],[18] Twenty Immature female rats (21 days old) received 1 mg/100g of testosterone enanthate (obtained from Iran Hormone Company, Tehran, Iran) subcutaneously daily for 35 days. After PCOS induction, the animals are divided into four groups containing five animals each group.
- Negative control group: normal saline recipient, for 28 days, orally.
- Positive control group: metformin 20 mg/kg/day for 28 days, by oral.
- Treatment group: Bryonia extract 30 mg/kg/day for 28 days, by oral (B1)
- Treatment group: Bryonia extract 60 mg/kg/day for 28 days, by oral (B2)
After treatment, the animals were subjected to biochemical, hormonal, and histopathological examination.
Measurement of follicle-stimulating hormone, glucose, low-density lipoprotein/ high-density lipoprotein, luteinizing hormone, and testosterone levels
On the last day of study, euthanasia was performed with an overdose of sodium pentobarbital. The blood samples were collected, and FSH, glucose, low-density lipoprotein/high-density lipoprotein (LDL/HDL), LH, and testosterone levels were measured. The serum levels of testosterone, FSH, and LH were measured by using the mouse enzyme-linked immunosorbent assay (ELISA) kit.[19]
Histological examination of ovaries
Histopathological studies were performed in ovarian tissues. Hematoxylin and eosin (H&E) staining following the method of Earl L. Parr[20] was used for slide preparation. The samples were then viewed and evaluated under a light microscope at ×40 magnification.
Data analysis
Data were statistically analyzed using Statistical Package for the Social Sciences (SPSS) software program, version 21.0. One-way analysis of the variance (ANOVA) and Tukey’s post hoc test were used to statistically analyze the data and compare the means of the groups. Results were reported as mean ± standard error of the mean (SEM). A value of P < 0.05 was considered significant.
| Results | | |
Effects of treatment on serum levels of hormones
The effect of B. dioica on hormones is shown in [Figure 1][Figure 2][Figure 3]. Testosterone levels were significantly decreased in metformin group (P < 0.001) as well as in B1 (P < 0.05) and B2 groups (P < 0.001) as compared with PCOS group. LH and FSH levels were significantly higher in the metformin group than in the PCOS group (P < 0.01). Bryonia dioica at 30 mg/kg showed a significantly greater level of FSH and LH than the PCOS group (P < 0.05). | Figure 1: Serum levels of testosterone in PCOS rats. *P < 0.05 and ***P < 0.001 vs. PCOS group. PCOS = polycystic ovary syndrome group, MET = metformin group, B1 = Bryonia extract group (30 mg/kg/day), B2 = Bryonia extract group (60 mg/kg/day)
Click here to view |
 | Figure 2: Serum levels of LH in PCOS rats. *P < 0.05 and **P < 0.01 vs. PCOS group. LH = luteinizing hormone, PCOS = polycystic ovary syndrome group, MET = metformin group, B1 = Bryonia extract group (30 mg/kg/day), B2 = Bryonia extract group (60 mg/kg/day)
Click here to view |
 | Figure 3: Serum levels of FSH in PCOS rats. *P < 0.05 and **P < 0.01 vs. PCOS group. FSH = follicle-stimulating hormone, PCOS = polycystic ovary syndrome group, MET = metformin group, B1 = Bryonia extract group (30 mg/kg/day), B2 = Bryonia extract group (60 mg/kg/day)
Click here to view |
Effect of Bryonia dioica on serum low-density lipoprotein and low-density lipoprotein/high-density lipoprotein ratio
As shown in [Figure 4] and [Figure 5], the LDL level was significantly decreased in B1 (P < 0.05) and B2 (P < 0.01) groups as compared to the PCOS group. In addition, the ratio of LDL/HDL markedly decreased in B1 (P < 0.05) and B2 (P < 0.01) groups. | Figure 4: Serum levels of LDL in PCOS rats. *P < 0.05 and **P < 0.01 vs. PCOS group. LDL = low-density lipoprotein, PCOS = polycystic ovary syndrome group, MET = metformin group, B1 = Bryonia extract group (30 mg/kg/day), B2 = Bryonia extract group (60 mg/kg/day)
Click here to view |
 | Figure 5: Serum levels of LDL/HDL in PCOS rats. *P < 0.05 and **P < 0.01 vs. PCOS group. LDL = low-density lipoprotein, HDL = high-density lipoprotein, PCOS = polycystic ovary syndrome group, MET = metformin group, B1 = Bryonia extract group (30 mg/kg/day), B2 = Bryonia extract group (60 mg/kg/day)
Click here to view |
Effect of Bryonia dioica on serum glucose
The effect of B. dioica on serum glucose is shown in [Figure 6]. The glucose levels were significantly decreased in B1 group as compared with the PCOS group (P < 0.001). Also serum glucose was markedly lower in B2 group than in the PCOS group (P < 0.0001). | Figure 6: Serum levels of glucose in PCOS rats. ***P < 0.001 and ****P < 0.0001 vs. PCOS group. PCOS = polycystic ovary syndrome
Click here to view |
Bryonia dioica restricted abnormal ovarian morphology in polycystic ovary syndrome rats
[Figure 7]A (normal group without induction of PCOS) shows too many corpus luteum (CL) cells––mature follicles (Graafian follicles) seen with normal granulosa cells. Numerous primary and secondary (antral) follicles. All cell’s structures were normal. In the PCOS group [Figure 7]B, no CL was seen. Follicular cyst with a thin layer of granulosa cells was also seen. Absence of primary follicles and mature follicles was seen. In metformin-treated group [Figure 7]C, pre-antral and primary follicles were present. The CL was seen. Granulosa cells went back to normal. | Figure 7: Morphological alterations in rat ovarian tissues. Rat ovarian tissues were stained with H&E, as explained in the “Materials and Methods” section, to observe morphological changes (×40). (A) Normal ovary. (B) PCOS group. (C) Metformin-treated group. (D) B1 (30 mg/kg/day)-treated group. (E) B2 (60 mg/kg/day)-treated group. H&E = hematoxylin and eosin, PCOS = polycystic ovary syndrome group, B1 = Bryonia extract group, B2 = Bryonia extract group
Click here to view |
In B1-treated group [Figure 7]D, relative improvement in structure of cells was observed as compared to PCOS group. The walls of the follicles and the granulosa cell layer were normal.
Graafian follicles contain ovum, and primary and secondary follicles were present, indicating normalization of the ovary. In the B2-treated group [Figure 7]E, mature follicles with normal granulosa layer were observable. Ovum, primary follicles, and CL were present.
| Discussion | | |
PCOS is an intricate metabolic and endocrine syndrome described by irregular ovulation and hyperandrogenism.[21],[22] In PCOS, increased production of androgens and estradiol is an early sign of abnormal steroidogenesis. Conversely, increased secretion of anti-Müllerian hormones (AMH) and LH, increased Gonadotropin-releasing hormone (GnRH) pulse rate, and decreased FSH levels are signs of the hypothalamic–pituitary–malfunctioning axis.[23] Treatment for this syndrome includes lifestyle changes, surgery, and taking some medicines, including clomiphene citrate and metformin.[24] Due to the unwanted effects of these drugs, the use of herbal medicines and their derivatives, which are less expensive, less invasive, and more valuable than other methods, has been considered.[25] As the identification of side effects of these medicines is momentous in the control of PCOS, in the various studies, the focus has been on the study of herbal medicines.[26] Having biologically active compounds and no significant side effects has caused special attention to be paid to the consumption of medicinal plants. For example, plants such as Red clover (Trifolium pratense), genistein, and soy have antiandrogenic properties due to the presence of specific phyto-estrogenic compounds such as biochanin A and daidzein, which can be used therapeutically in patients with PCOS.[27],[28],[29] It has been shown that flavonoid compounds in plants with antioxidant activity can reduce oxidative stress in ovarian tissue and reduce the number of cystic follicles in this tissue.[30] Plant antioxidants also reduced the effects of hyperandrogenism following PCOS induction in mice.[31] The antiandrogenic properties of plants also reduce the secretion of androgens by inducing negative feedback on LH. Decreased androgens lead to less LH synthesis and reduce the dominant effect of LH on FSH, which in turn may be a reason for the restart of the natural pathway of sex cycle hormones and the occurrence of ovulation in PCOS.[32],[33]Bryonia dioica is one of the traditional medicines, which is widely used in different parts of the world, especially in Western Europe. The root of B. dioica is enriched in bioactive compounds such as kempferol 3, 7-di-O-rhamnoside, and polyphenols that show pharmacological effects including antioxidant activity, analgesia, hepato-protective, antihypercholesterolemia and hyperglycemia, and improving fertility disorders.[34],[35],[36] Previous studies have shown that aqueous extract of B. dioica in low concentrations (50 µg/mL) in MDA MB-231 cells can cause apoptosis and stop the cell cycle in the G2/M phase.[37] This study intended to establish an animal model of PCOS using DHEA. DHEA was used in some studies to induce PCOS in rats. Histological analysis of ovarian tissues clearly showed the treated cystic follicles.[37],[38] The first androgen to appear in the blood of adult women is DHEA,[39] which also increases in 25% of PCOS cases.[40] DHEA administration during fertility causes complications of PCOS, such as menstrual disorders and polycystic ovaries.[41],[42] The results of our research showed that B. dioica normalized the effect of testosterone on ovary of rats as well as the serum levels of LH and FSH. A study by Chauhan and Dixit[43] found that the ethanolic extract of B. laciniosa Linn. seeds increased FSH and LH levels. Our data showed that the levels of FSH and LH in the treatment group were improved compared with untreated group. In previous studies, the effect of B. laciniosa on glucose homeostasis has been reported in the streptozotocin (STZ) diabetic model of rats. Therefore, the antidiabetic effect of this plant can be used in the treatment of diabetic patients.[13],[44] In our study, glucose levels were significantly reduced in the B. dioica groups (30 and 60 mg/kg) when compared with the PCOS group. Significant reductions in LDL and LDL/HDL ratio were observed in rats treated with the ethanolic extract of B. laciniosa and the saponin component of B. laciniosa.[45] In this study, B. dioica (60 mg/kg) decreased LDL levels in PCOS rats and the LDL/HDL ratio was decreased in all treatment groups compared to the PCOS group.
| Conclusion | | |
According to the study’s data, B. dioica showed the protective effect on PCOS rats and normalized the hormones, glucose, LDL, and LDL/HDL ratio, which are abnormal in PCOS rats. Moreover, B. dioica has an improvement effect on the symptoms and markers of PCOS and fertility but more researches to get precise results are needed.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
| References | | |
| 1. | DuRant E, Leslie NS Polycystic ovary syndrome: A review of current knowledge. J Nurse Pract 2007;3:180-5.  |
| 2. | Toscani MK, Mario FM, Radavelli-Bagatini S, Spritzer PM Insulin resistance is not strictly associated with energy intake or dietary macronutrient composition in women with polycystic ovary syndrome. Nutr Res 2011;31:97-103. |
| 3. | Kriedt KJ, Alchami A, Davies MC PCOS: Diagnosis and management of related infertility. Obstet Gynaecol Reprod Med 2019;29:1-5. |
| 4. | Franks S Polycystic ovary syndrome. N Engl J Med 1995;333:853-61. |
| 5. | Glueck CJ, Phillips H, Cameron D, Sieve-Smith L, Wang P Continuing metformin throughout pregnancy in women with polycystic ovary syndrome appears to safely reduce first-trimester spontaneous abortion: A pilot study. Fertil Steril 2001;75:46-52. |
| 6. | Marshall K Polycystic ovary syndrome: Clinical considerations. Altern Med Rev 2001;6:272-92. |
| 7. | Devi K, Santhi M, Umadevi U Phytochemical analysis of selected wound healing medicinal plants. Int J Pharm Sci Res 2017;8:852-5. |
| 8. | Adlercreutz H, Mazur W Phyto-oestrogens and western diseases. Ann Med 1997;29:95-120. |
| 9. | Hylands PJ, Mansour E-SS New cucurbitacin derivatives from Bryonia dioica Jacq. J Chem Soc Perkin Trans 1983;1:2821-5. |
| 10. | Benarba B, Elmallah A, Pandiella A Bryonia dioica aqueous extract induces apoptosis and G2/M cell cycle arrest in MDA-MB 231 breast cancer cells. Mol Med Rep 2019;20:73-80. |
| 11. | Akihisa T, Kimura Y, Kokke WC, Itoh T, Tamura T. Eight novel sterols from the roots of Bryonia dioica JACQ. Chem Pharm Bull1996;44:1202-7. |
| 12. | Hylands PJ, Kosugi J Bryonoside and bryoside: New triterpene glycosides from Bryonia dioica. Phytochemistry1982;16:1379-84. |
| 13. | Chekroun E, Bechiri A, Azzi R, Adida H, Benariba N, Djaziri R. Antidiabetic activity of two aqueous extracts of two Cucurbitaceae: Citrullus colocynthis and Bryonia dioica. Phytothérapie 2017;15:57-66. |
| 14. | Chitra V, Dhivya DP Role of herbal in management of polycystic ovarian syndrome and its associated symptoms. Int J Herb Med 2017;5:125-31. |
| 15. | Lee J, Lim SC, Lyu AR, Bae JG, Kang SS, Kim JC, et al. Effects of red ginseng total saponin on the polycystic ovaries in rats. KSBB J 2004;19:433-6. |
| 16. | Ben Abdessamad I, Bouhlel I, Chekir-Ghedira L, Krifa M Antitumor effect of Bryonia dioica methanol extract: In vitro and in vivo study. Nutr Cancer 2020;72:747-56. |
| 17. | Osuka S, Nakanishi N, Murase T, Nakamura T, Goto M, Iwase A, et al. Animal models of polycystic ovary syndrome: A review of hormone-induced rodent models focused on hypothalamus-pituitary-ovary axis and neuropeptides. Reprod Med Biol 2019;18:151-60. |
| 18. | Karateke A, Dokuyucu R, Dogan H, Ozgur T, Tas ZA, Tutuk O, et al. Investigation of therapeutic effects of erdosteine on polycystic ovary syndrome in a rat model. Med Princ Pract 2018;27:515-22. |
| 19. | van Casteren JI, Schoonen WG, Kloosterboer HJ Development of time-resolved immunofluorometric assays for rat follicle-stimulating hormone and luteinizing hormone and application on sera of cycling rats. Biol Reprod 2000;62:886-94. |
| 20. | Parr EL Histological examination of the rat ovarian follicle wall prior to ovulation. Biol Reprod 1974;11:483-503. |
| 21. | Sun J, Jin C, Wu H, Zhao J, Cui Y, Liu H, et al. Effects of electro-acupuncture on ovarian p450arom, P450c17α and mRNA expression induced by letrozole in PCOS rats. PLoS One 2013;8:e79382. |
| 22. | Yang H, Lee SY, Lee SR, Pyun BJ, Kim HJ, Lee YH, et al. Therapeutic effect of Ecklonia cava extract in letrozole-induced polycystic ovary syndrome rats. Front Pharmacol 2018;9:1325. |
| 23. | Bednarska S, Siejka A The pathogenesis and treatment of polycystic ovary syndrome: What’s new? Adv Clin Exp Med 2017;26:359-67. |
| 24. | Sirmans SM, Pate KA Epidemiology, diagnosis, and management of polycystic ovary syndrome. Clin Epidemiol 2014;6:1-13. |
| 25. | Beazley KE, Nurminskaya M Effects of dietary quercetin on female fertility in mice: Implication of transglutaminase 2. Reprod Fertil Dev 2016;28:974-81. |
| 26. | Pachiappan S, Matheswaran S, Saravanan PP, Muthusamy G. Medicinal plants for polycystic ovary syndrome: A review of phytomedicine research. Int J Herb Med 2017;5:78-80. |
| 27. | Rajan RK, M SS, Balaji B Soy isoflavones exert beneficial effects on letrozole-induced rat polycystic ovary syndrome (PCOS) model through anti-androgenic mechanism. Pharm Biol 2017;55:242-51. |
| 28. | Kargozar R, Azizi H, Salari R A review of effective herbal medicines in controlling menopausal symptoms. Electron Physician 2017;9:5826-33. |
| 29. | Amanat S, Ashkar F, Eftekhari MH, Tanideh N, Doaei S, Gholamalizadeh M, et al. The effect of genistein on insulin resistance, inflammatory factors, lipid profile, and histopathologic indices in rats with polycystic ovary syndrome. Korean J Fertil Steril 2021;48:236-44. |
| 30. | Yilmaz N, Inal HA, Gorkem U, Yilmaz S, Sargin A, Turkkani A. Follicular fluid total antioxidant capacity in patient with PCOS. Fertil Steril 2013;100:654-7. |
| 31. | Abasian Z, Rostamzadeh A, Mohammadi M, Hosseini M, Rafieian-Kopaei M. A review on role of medicinal plants in polycystic ovarian syndrome: Pathophysiology, neuroendocrine signaling, therapeutic status and future prospects. Middle East Fertil Soc J 2018;23:255-62. |
| 32. | Armanini D, Mattarello MJ, Fiore C, Bonanni G, Scaroni C, Sartorato P, et al. Licorice reduces serum testosterone in healthy women. Steroids 2004;69:763-6. |
| 33. | Ashkar F, Rezaei S, Salahshoornezhad S, Vahid F, Gholamalizadeh M, Dahka SM, et al. The role of medicinal herbs in treatment of insulin resistance in patients with polycystic ovary syndrome: A literature review. Biomol Concepts 2020;11:57-75. |
| 34. | Jasiem TM, Eldalawy R, Alnaqqash ZAE Pharmacological activities and chemical constituents and of Bryonia dioica L.: A review. Indian J Public Health 2020;11:2185-90. |
| 35. | Khamees AH, Kadhim EJ, Sahib HB, Mutlag SH. In vitro analysis of antioxidant and antimicrobial activity of Iraqi Bryonia dioica. Int J Pharm Sci Rev Res 2017;43:248-52. |
| 36. | Benarba B Ethnomedicinal study of Bryonia dioica: A plant used as anti-breast cancer herbal therapy in North West Algeria. J Med Herb Ethnomed 2015;1:113-5. |
| 37. | Wang MX, Yin Q, Xu X A rat model of polycystic ovary syndrome with insulin resistance induced by letrozole combined with high fat diet. Med Sci Monit 2020;26:e922136. |
| 38. | Abhari SM, Khanbabaei R, Roodbari NH, Parivar K, Yaghmaei P. Curcumin-loaded super-paramagnetic iron oxide nanoparticle affects on apoptotic factors expression and histological changes in a prepubertal mouse model of polycystic ovary syndrome-induced by dehydroepiandrosterone: A molecular and stereological study. Life Sci 2020;249:117515-24. |
| 39. | Roy S, Mahesh VB, Greenblatt RB Effect of dehydroepiandrosterone and delta4-androstenedione on the reproductive organs of female rats: Production of cystic changes in the ovary. Nature 1962;196:42-3. |
| 40. | Azziz R, Carmina E, Dewailly D, Diamanti-Kandarakis E, Escobar-Morreale HF, Futterweit W, et al. The androgen excess and PCOS society criteria for the polycystic ovary syndrome: The complete task force report. Fertil Steril2009;91:456-88. |
| 41. | Seow KM, Ting CH, Huang SW, Ho LT, Juan CC The use of dehydroepiandrosterone-treated rats is not a good animal model for the study of metabolic abnormalities in polycystic ovary syndrome. Taiwan J Obstet Gynecol 2018;57:696-704. |
| 42. | Jang M, Lee MJ, Lee JM, Bae CS, Kim SH, Ryu JH, et al. Oriental medicine Kyung-Ok-Ko prevents and alleviates dehydroepiandrosterone-induced polycystic ovarian syndrome in rats. PLoS One 2014;9:e87623. |
| 43. | Chauhan NS, Dixit VK Effects of Bryonia laciniosa seeds on sexual behaviour of male rats. Int J Impot Res 2010;22:190-5. |
| 44. | Patel SB, Santani D, Patel V, Shah M Anti-diabetic effects of ethanol extract of Bryonia laciniosa seeds and its saponins rich fraction in neonatally streptozotocin-induced diabetic rats. Pharmacognosy Res 2015;7:92-9. |
| 45. | Patel SB, Santani D, Shah M, Patel VS Anti-hyperglycemic and anti-hyperlipidemic effects of Bryonia laciniosa seed extract and its saponin fraction in streptozotocin-induced diabetes in rats. J Young Pharm 2012;4:171-6. |
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]
|
Search Pubmed for