Identification of Steroidogenic Components Derived From Gardenia jasminoides Ellis Potentially Useful for Treating Postmenopausal Syndrome
In addition, the possibility of toxicity of GJE was ruled out and four probable active compounds were predicted.
Among the nine compounds, four of them established by network pharmacology, validating the use of network pharmacology to predict active compounds.
The stimulation of estrogen production by rat ovarian granulosa cells under the influence of the three compounds underwent a decline when the follicle-stimulating hormone receptor( FSHR) was blocked by antibodies against the receptor, indicating the involvement of FSHR in the estradiol-stimulating activity of the three compounds.
Estrogen-stimulating principles have been demonstrated to relieve postmenopausal disorder effectively. Gardenia jasminoides Ellis( GJE) is an herbal medicine possessing multiple pharmacological consequences on human health with low toxicity. However, the therapeutic effects of GJE on the management of postmenopausal syndrome and its mechanism of action have not been fully elucidated. In this study, network pharmacology-based approaches were employed to examine steroidogenesis under the influence of GJE . In addition, the possibility of toxicity of GJE was ruled out and four probable active compounds were predicted. In parallel, a chromatographic fraction of GJE with estrogen-stimulating effect was identified and nine major compounds were isolated from this active fraction. Among the nine compounds, four of them established by network pharmacology, validating the use of network pharmacology to predict active compounds. Then the phenotypic approaches were utilized to verify that rutin, chlorogenic acid( CGA) and geniposidic acid( GA) exerted an estrogen-stimulating consequence on ovarian granulosa cells. Furthermore, the results of target-based approaches indicated that rutin, CGA, and GA could up-regulate the FSHR-aromatase pathway in ovarian granulosa cells. The stimulation of estrogen production by rat ovarian granulosa cells under the influence of the three compounds underwent a deterioration when the follicle-stimulating hormone receptor( FSHR) was blocked by antibodies against the receptor, indicating the involvement of FSHR in the estradiol-stimulating activity of the three compounds. The effects of the three compounds on estrogen biosynthesis- related gene expression level were farther confirmed by Western blot assay. Importantly, the MTT outcomes showed that exposure of breast cancer cells to the three compounds resulted in reduction of cell viability, demonstrating the cytotoxicity of the three compounds. Collectively, rutin, chlorogenic acid and geniposidic acid may contribute to the therapeutic potential of GJE for the treatment of postmenopausal syndrome.
Natural menopause is classified as permanent cessation of menstruation, induced by ovarian follicular failing and ovarian hormone instability( Burger et al ., 2007 ). Due to a global population aging, there will be one billion girls over the age of 60 years by the year 2050( Hoga et al ., 2015 ). Currently, more than 50% of women in the world are afflicted with postmenopausal syndrome at the climacteric stage( Su et al ., 2013; Dalal and Agarwal, 2015 ). It is widely accepted that estrogen deprivation plays an important role in the postmenopausal disorder( World Health Organization, 1996; Greendale et al ., 1998; Constantine and Pickar, 2003 ). In the climacteric period, the decline of estrogen may induce depression( Kaufert et al ., 1992; Avis et al ., 1994; Al-Safi and Santoro, 2014; Citraro et al ., 2015 ), memory loss( Devi et al ., 2005; Weber and Mapstone, 2009 ), bone resorption( Hernandez et al ., 2003; Society, 2006 ), metabolic disorder( Carr, 2003; Janssen et al ., 2008 ), and colorectal cancer( Al-Azzawi and Wahab, 2002; Barzi et al ., 2013 ). The physiological problems caused by estrogen deficiency adversely influence the quality of life of modern people and have become a substantial public health onu. To relieve the reduced level of estrogen, women in climacterium usually opt to undergo hormone replacement therapy( HRT)( Barnabei et al ., 2002 ). However, a WHO analyse has established that HRT can significantly heighten the risk of endometrial cancer, breast cancer and gallbladder cancers in climacteric females( Nelson et al ., 2002; Davey, 2013; Chuffa et al ., 2016 ). In addition, HRT is usually accompanied by considerable untoward side effect, including vaginal bleeding, genital annoyance and headache( Clarke et al ., 2002 ). Therefore, a safe and effective treatment of postmenopausal syndrome is necessitated.
As alternative solutions therapy for postmenopausal disorder, herbal medicine has a long history of a thousand years and a wide range of applications for improving women’s health( Johnston, 1997; Feng and Cao, 2010 ). In the US, over 80% of the physicians suggest that their patients alleviate postmenopausal disorder with herbal medicine ( Meisler, 2003 ). In China and other Asian countries, herbal medication has been extensively and chronically deployed to alleviate postmenopausal syndrome, due to its well-known safety and efficacy( Scheid, 2007; Liu et al ., 2008; Chen et al ., 2010; Scheid et al ., 2010 ). Among the selected herbal medication, Gardenia jasminoides Ellis( GJE) is a potential candidate for the therapy of gynecological disorders( Yang et al ., 2016 ). GJE has a wide range of pharmacological consequences, including anti-allergy( Sung et al ., 2014 ), anti-oxidative, anti-atherosclerotic, anti-platelet aggregating, anti-hypertensive activities, and so on( Liu et al ., 2013 ). Nevertheless, the effects of GJE on the management of climacterium have rarely been reported. Lately, several surveys reported that the fractions of GJE and the major compounds in GJE could display antidepressant activities in rodent models( Cai et al ., 2015; Zhang et al ., 2015; Ren et al ., 2016; Wang et al ., 2016 ). Additionally, previous examines indicated that GJE and its active components could improve memory and learning ability, and protect the neurons in animals with brain trauma( Sheng et al ., 2006; Chen et al ., 2015; Zhang et al ., 2016 ). Genipin, a major phytoconstituent of GJE, is a candidate for the treatment of osteoporosis( Hoon Lee et al ., 2014 ). GJE was also able to attenuate metabolic syndrome with a combination of other herbal drugs in estrogen-deficient rats( Yang et al ., 2016 ). Moreover, the components of GJE exerted suppressive consequences on colon cancer cells and breast cancer cells( Kim et al ., 2012; Oliveira et al ., 2017 ). Gardenia oil also augmented plasma estradiol levels( Li et al ., 2013 ). Collectively, GJE can be regarded as a promising nominee for the treatment of estrogen deprivation. However, neither systematic mechanistic analyzes of GJE related to estrogen deprivation nor the estrogen-stimulating effects of GJE have been reported. Hence, it was hypothesized that there are several active compounds derived from GJE that are endowed with the ability of stimulating estrogen biosynthesis in ovarian granulosa cells.
In this study, network and systemic pharmacological analysis was used to identify the therapeutic role of GJE in the treatment of postmenopausal syndrome and in the prevention of health risks of HRT. Firstly, the possible bioactive compounds of GJE were predicted by network pharmacology. Afterwards, the fractions of GJE were isolated by HPLC and the bioactive fraction of GJE that could be adopted to treat postmenopausal syndrome was investigated by using the estradiol assay. The major components were then removed from the bioactive fraction and converged with the probable active ingredients predicted by network pharmacology. Among these components, phenotypic( on the basis of the estradiol assay) and target-based( on the basis of the molecular docking analysis, Western blot assay and FSHR inhibition assay) drug-screening principles were applied to screen bioactive compounds with estrogen-stimulating impacts( Hughes et al ., 2011; Lu et al ., 2016 ). Breast cancer cells were also are applied to pre-evaluate the cancer danger of GJE active ingredients. The workflow to explore the bioactive compounds is illustrated in Figure 1. Results obtained from this study disclose the estrogen-stimulating action of GJE, which further supports the clinical utilize of GJE to attenuate postmenopausal syndrome.