Anti-Müllerian hormone participates in ovarian granulosa cell damage due to cadmium exposure by negatively regulating stem cell factor
Introduction
Cadmium (Cd) a key heavy metal environmental toxicant that ranked among the top seven on the 2017 Agency for Toxic Substances and Disease Registry (ATSDR) Priority List of Hazardous Substances [1]. Since 1950, global cadmium production has been increasing year by year, reaching 23 000 tons in 2015 [2]. The average content of Cd in the soils of the United States, Europe, England and Wales, and China is 0.34 mg/kg, 0.18 mg/kg, 0.33 mg/kg, 0.24 mg/kg, respectively [3]. According to a study in Bangladesh, the highest concentration of Cd was 17 mg/kg in the Turag River [2].
Cadmium enters the body through three primary sources: diet, smoking, and air pollution [4]. Among the various diets worldwide, rice is the most significant contributor to Cd intake in the diet, although the extent of its contribution depends on the country [5]. In various human diets, rice (Oryza sativa) is the largest contributor to Cd intake in the diet of ordinary non-smokers, resulting in higher Cd content in the diet [6,7]. Compared to the United States and European countries, Asian countries generally have larger rice intakes [5]. In China, the rapid industrialization of the past 30 years and insufficient environmental protection have caused the soil to be seriously polluted by heavy metals and metalloids. The accumulation of Cd was continuously increasing during the period from 2005 to 2017 in china [3]. A survey in the southern part of Jiangsu Province, China, showed that about 14.89 % of the cadmium in the soil was transferred to rice, and up to 3.19 % of the rice was transferred from cadmium to the human body [8]. Cadmium has acute and chronic effects on health. Chronic Cd exposure can adversely affect the growth and development of the liver [9], kidneys [10,11], bones [12], cardiovascular system [13], and fetals growth and development [14], and increase the risk of death from various cancers [4]. It is known that Cd exposure can also affect the female reproductive system. The study reported that Cd retention is generally higher in women compared to men [15], and the Cd-induced Itai-Itai ailment was mostly observed in women [16,17]. Cadmium tends to accumulate in ovarian granulosa cells. Cd2+inhibited progesterone synthesis in cultured granulosa cells from both rats and humans. Cd induce endocrine disruption at 5–20 μM concentrations in cultured human trophoblasts were observed [18].
The female reproductive system is an extremely rich environment with many different cell types. The mural granulosa cells lying on the basal membrane of the follicular wall and cumulus granulosa cells (CCs) surrounding oocyte [19]. Granulosa cells (GCs) act as essential mediators in ovarian follicle development and produce necessary growth factors and specific proteins (e,g., AMH, SCF, activin, inhibin, IGF-1). The proliferation of GCs triggers the follicular growth and oocyte maturation [20]. Anti-Müllerian hormone (AMH) is a dimeric glycoprotein member of the transforming growth factor-beta (TGF-β) superfamily [21]. In the ovary, the granulosa cells that develop follicles in early development secrete AMH, which inhibit the initial recruitment of primordial follicles and regulate the growth and development of follicles [22]. Stem cell factor (SCF), also known as a Kit ligand (KITLG), is the ligand for the tyrosine-protein kinase receptor (c-kit) [23]. In the ovary, SCF promotes primordial germ cell migration, proliferation, and differentiation [24,25]. It is evident that even today, there are large gaps in knowledge with regard to the assessment of the reproductive effects caused by Cd. For example, What is the minimum concentration of cadmium that affects the female reproductive system? How does low-dose long-term exposure affect the female reproductive system? Why do high- and low-dose cadmium exposures sometimes cause opposite effects? What is the specific mechanism of cadmium affecting female reproduction? Therefore, although the results of many studies have shown significant effects on reproduction, specific mechanisms of action need to be studied.
The objective of this in vitro study was to examine the changes in the secretion of the Anti-Müllerian hormone and Stem cell factor after the addition of 5–20 μM Cadmium chloride to ovarian granulosa cells. The study also aimed at ascertaining the apoptotic potential of ovarian granulosa cells in relation to the possible involvement of the Anti-Müllerian hormone and Stem cell factor in ovarian functions.
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Animals
The animals were provided with food and water at random, and reared in accordance with the guidelines for the care and use of experimental animals. Sprague Dawley (SD) rats (21 days old) (animal license: SCXK 2012-0002, Shanghai Slark Experimental Animal Co., Ltd., China) were placed in an environment-controlled room under 14 h of light and 10 h of light Inside the dark photoperiod. Keep temperature at the 25 ± 2 °C and humidity at 50 %. All animal experiments were approved by the Animal
The viability of ovarian granulosa cells
The viability of rat ovarian granulosa cells treated with different concentrations of cadmium chloride (5 μM, 10 μM, and 20 μM) decreased significantly when compared with that of the control group (0 μM) at 12 h, 24 h and 36 h (Fig. 1A, P < 0.05). In the 20 μM cadmium chloride-treated group, granulosa cell viability was significantly inhibited in the 24 h and 36 h groups compared with that of the 12 h group, and this effect was time-dependent (Fig. 1A). The results also showed that when the
Discussion
Both AMH and SCF signaling pathways are involved in the reproductive effects caused by Cd. This study demonstrated that, in the mouse ovary, Cd promotes granulosa cell apoptotic and that this effect is at least in part mediated through regulating AMH signaling pathway. This study also demonstrated that AMH signaling is involved in Cd action through SCF activation. The signaling pathways may play a role in the reproductive effects caused by environmental and occupational exposures to Cd.
Cadmium
Declaration of Competing Interest
The authors declare that they have no known competing financialinterestsor personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
This study was supported by the National Natural Science Foundation of China (Grant No. 81373027, 81673212), the Natural Science Foundation of Fujian Province (2016J01359), and the Young Research Foundation of Fujian Provincial Health and Family Planning Commission (2017-1-66).
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These authors contributed equally to the work.