Effect of Vitex agnus-castus ethanolic extract on hypothalamic KISS-1 gene expression in a rat model of polycystic ovary syndrome

Document Type : Original Research Article


Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran


Objective: Polycystic ovary syndrome (PCOS) is an endocrine system disruption that affects 6-10% of women. Some studies have reported the effect of Vitex agnus-castus (Vitagnus) on the hypothalamic-pituitary-gonad axis (HPG). This study was conducted to investigate Vitagnus effect on the expression of kisspeptin gene in a rat model of PCOS.
Materials and Methods: Thirty-two female rats were distributed into: control, Vitagnus-treatment (365 mg/kg for 30 days), PCOS (Letrozole for 28 days) and PCOS animals treated with Vitagnus (30 days of Vitagnus after PCOS induction). At the end of the treatments, serum and ovaries were collected for analysis. Expression level of KISS-1 gene in the hypothalamus was investigated, using Real-Time-PCR.
Results: In the PCOS group compared to control, FSH, progesterone and estradiol levels were decreased, whereas testosterone and LH levels were significantly increased. No significant changes were observed in the Vitagnus-treated animals in compare to control.  However, Vitagnus treatment in the PCOS group, resulted in a raise in progesterone, estrogen and FSH levels and a reduction in the levels of testosterone and LH. Quantitative gene expression analysis showed that PCOS induction resulted in over-expression of KISS-1 gene, however, Vitagnus treatment reduced this up-regulated expression to normal level.
Conclusion: In conclusion, our results indicated that Vitagnus extract inhibited downregulation of KISS-1 gene in the hypothalamus of PCOS rats. Because of the master role of kisspeptin in adjusting the HPG axis, Vitagnus is likely to show beneficial effects in the treatment of PCOS via regulation of kisspeptin expression. This finding indicates a new aspect of Vitagnus effect and may be considered in its clinical applications.


Aliabadi E, Namavar MR, Mortezaee K, Toolee
H, Keshtgar S, Mirkhani H, Akbari M,
Rastegar T, Solhjoo S. 2017. Kisspeptin
expression features in the arcuate and
anteroventral periventricular nuclei of
hypothalamus of letrozole-induced
polycystic ovarian syndrome in rats. Arch
Gynecol Obstet, 296: 957-963.
Allahtavakoli M, Honari N, Pourabolli I,
Kazemi Arababadi M, Ghafarian H,
Roohbakhsh A, Esmaeili Nadimi A,
Shamsizadeh A. 2015. Vitex Agnus Castus
Extract Improves Learning and Memory and
Increases the Transcription of Estrogen
Receptor alpha in Hippocampus of Ovarie
ctomized Rats. Basic Clin Neurosci, 6: 185-
Arentz S, Abbott JA, Smith CA, Bensoussan A.
2014a. Herbal medicine for the management
of polycystic ovary syndrome (PCOS) and
associated oligo/amenorrhoea and
hyperandrogenism; a review of the
laboratory evidence for effects with
corroborative clinical findings. BMC
Complementary and Alternative Medicine,
14: 511-580.
Badawy A, Elnashar A. 2011. Treatment
options for polycystic ovary syndrome. Int J
Womens Health, 3: 25-35.
Casper RF, Mitwally MF. 2011. Use of the
aromatase inhibitor letrozole for ovulation
induction in women with polycystic ovarian
syndrome. Clin Obstet Gynecol, 54: 685-95.
Chen SN, Friesen JB, Webster D, Nikolic D,
Van Breemen RB, Wang ZJ, Fong HH,
Farnsworth NR, Pauli GF. 2011.
Phytoconstituents from Vitex agnus-castus
fruits. Fitoterapia, 82: 528-33.
Feyzollahi et al.
AJP, Vol. 11, No. 3, May-Jun 2021 300
Clarkson J, D'anglemont De Tassigny X,
Colledge WH, Caraty A, Herbison AE.
2009. Distribution of kisspeptin neurones in
the adult female mouse brain. J
Neuroendocrinol, 21: 673-82.
Dubois SL, Acosta-Martinez M, Dejoseph MR,
Wolfe A, Radovick S, Boehm U, Urban JH,
Levine JE. 2015. Positive, but not negative
feedback actions of estradiol in adult female
mice require estrogen receptor alpha in
kisspeptin neurons. Endocrinol, 156: 1111-
Gorkem U, Togrul C, Arslan E, Sargin Oruc A,
Buyukkayaci Duman N. 2018. Is there a role
for kisspeptin in pathogenesis of polycystic
ovary syndrome? Gynecol Endocrinol, 34:
Gottsch M, Cunningham MJ, Smith JT, Popa
SM, Acohido BV, Crowley WF, Seminara
S, Clifton DK, Steiner RA. 2004. A role for
kisspeptins in the regulation of gonadotropin
secretion in the mouse. Endocrinol, 145:
Heskes AM, Sundram TC, Boughton BA,
Jensen NB, Hansen NL, Crocoll C, Cozzi F,
Rasmussen S, Hamberger B. Hamberger B.
2018. Biosynthesis of bioactive diterpenoids
in the medicinal plant Vitex agnus‐castus.
The Plant Journal, 93: 943-958.
Hoeger KM, Legro RS, Welt CK. 2014. A
patient's guide: polycystic ovary syndrome
(PCOS). J Clin Endocrinol Metab, 99: 35A36A.
Kafali H, Iriadam M, Ozardalı I, Demir N.
2004. Letrozole-induced polycystic ovaries
in the rat: a new model for cystic ovarian
disease. Archives of Medical Research, 35:
Kakadia N, Patel P, Deshpande S, Shah G.
2019. Effect of Vitex negundo L. seeds in
letrozole induced polycystic ovarian
syndrome. J Tradit Complement Med, 9:
Katulski K, Podfigurna A, Czyzyk A,
Meczekalski B, Genazzani AD. 2018.
Kisspeptin and LH pulsatile temporal
coupling in PCOS patients. Endocrine, 61:
Kauffman AS, Clifton DK, Steiner RA. 2007a.
Emerging ideas about kisspeptin- GPR54
signaling in the neuroendocrine regulation
of reproduction. Trends Neurosci, 30: 504-
Kauffman AS, Clifton DK, Steiner R.A. 2007b.
Emerging ideas about kisspeptin–GPR54
signaling in the neuroendocrine regulation
of reproduction. Trends in Neurosciences,
30: 504-511.
Kondo M, Osuka S, Iwase A, Nakahara T, Saito
A, Nakamura T, Goto M, Kotani T,
Kikkawa F. 2016. Increase of kisspeptinpositive cells in the hypothalamus of a rat
model of polycystic ovary syndrome.
Metabolic Brain Disease, 31: 673-681.
Liu J, Burdette J, Sun Y, Deng S, Schlecht S,
Zheng W, Nikolic D, Mahady G, Van
Breemen R, Fong H. 2004a. Isolation of
linoleic acid as an estrogenic compound
from the fruits of Vitex agnus-castus
L.(chaste-berry). Phytomedicine, 11: 18-23.
Liu J, Burdette JE, Sun Y, Deng S, Schlecht
SM, Zheng W, Nikolic D, Mahady G, Van
Breemen RB, Fong HH, Pezzuto JM, Bolton
JL, Farnsworth NR. 2004b. Isolation of
linoleic acid as an estrogenic compound
from the fruits of Vitex agnus-castus L.
(chaste-berry). Phytomedicine, 11: 18-23.
Liu Z. 2008. Preparation of botanical samples
for biomedical research. Endocr Metab
Immune Disord Drug Targets, 8: 112-21.
Lizneva D, Suturina L, Walker W, Brakta S,
Gavrilova-Jordan L, Azziz R. 2016. Criteria,
prevalence, and phenotypes of polycystic
ovary syndrome. Fertil Steril, 106: 6-15.
Mccartney Ch R, Marshall JC. 2016. Polycystic
Ovary Syndrome. N Engl J Med, 375: 1398-
Nasri S, Oryan S, Rohani AH, Amin GR. 2007.
The effects of Vitex agnus castus extract and
its interaction with dopaminergic system on
LH and testosterone in male mice. Pak J Biol
Sci, 10: 2300-7.
Radosh L. 2009. Drug treatments for polycystic
ovary syndrome. Am Fam Physician, 79:
Romero-Ruiz A, Skorupskaite K, Gaytan F,
Torres E, Perdices-Lopez C, Mannaerts BM,
Qi S, Leon S, Manfredi-Lozano M, LopezRodriguez C, Avendano MS, SanchezGarrido MA,Vazquez MJ, Pinilla L, Van
Duin M, Kohout TA, Anderson RA, TenaSempere M. 2019. Kisspeptin treatment
induces gonadotropic responses and rescues
ovulation in a subset of preclinical models
and women with polycystic ovary
syndrome. Hum Reprod, 34: 2495-2512.
Russo M, Galletti GC. 1995. Medicinal
properties and chemical composition of
Vitex agnus-castus L.: a review.
International Symposium on Medicinal and
Effect of Vitagnus on hypothalamic KISS-1 gene
AJP, Vol. 11, No. 3, May-Jun 2021 301
Aromatic Plants 426: 105-112.
Sun J, Jin C, Wu H, Zhao J, Cui Y, Liu H, Wu
L, Shi Y, Zhu B. 2013. Effects of electroacupuncture on ovarian P450arom,
P450c17α and mRNA expression induced
by letrozole in PCOS rats. PLoS One, 8,
Tang R, Ding X, Zhu J. 2019. Kisspeptin and
Polycystic Ovary Syndrome. Front
Endocrinol (Lausanne), 10: 298-305.
Tena-Sempere M. 2006. The roles of
kisspeptins and G protein-coupled receptor54 in pubertal development. Current
Opinion in Pediatrics, 18: 442-447.
Trevisan CM, Montagna E, De Oliveira R,
Christofolini DM, Barbosa CP, Crandall
KA, Bianco B. 2018. Kisspeptin/GPR54
System: What Do We Know About Its Role
in Human Reproduction? Cell Physiol
Biochem, 49: 1259-1276.
Umayal B, Jayakody SN, Chandrasekharan
NV, Wijesundera WS, Wijeyaratne CN.
2019. Polycystic ovary syndrome (PCOS)
and kisspeptin - A Sri Lankan study. J
Postgrad Med, 65: 18-23.
Van Die MD, Burger HG, Teede HJ, Bone KM.
2013. Vitex agnus-castus extracts for female
reproductive disorders: a systematic review
of clinical trials. Planta Medica, 79: 562-
Witchel SF, Tena-Sempere M. 2013. The Kiss1
system and polycystic ovary syndrome:
lessons from physiology and putative
pathophysiologic implications. Fertil Steril,
100: 12-22.
Yang H, Kim HJ, Pyun BJ, Lee HW. 2018.
Licorice ethanol extract improves symptoms
of polycytic ovary syndrome in Letrozoleinduced female rats.