Shahzad Huma, Giribabu Nelli, Karim Kamarulzaman, Kassim Normadiah M, Muniandy Sekaran, Salleh Naguib
Department of Physiology, Faculty of Medicine, University of Malaya, Lembah Pantai, Kuala Lumpur, Malaysia.
Department of Anatomy, Faculty of Medicine, University of Malaya, Lembah Pantai, Kuala Lumpur, Malaysia.
PLoS One. 2017 Mar 2;12(3):e0172765. doi: 10.1371/journal.pone.0172765. eCollection 2017.
Dysregulation of uterine fluid environment could impair successful reproduction and this could be due to the effect of environmental estrogens. Therefore, in this study, effect of quercetin, an environmental estrogen on uterine fluid and electrolytes concentrations were investigated under sex-steroid influence. Ovariectomised adult female Sprague-Dawley rats were given 10, 50 or 100mg/kg/day quercetin subcutaneously with 17-β estradiol (E) for seven days or three days E, then three days E plus progesterone (P) (E+P) treatment. Uterine fluid secretion rate, Na+, Cl- and HCO3- concentrations were determined by in-vivo perfusion. Following sacrifice, uteri were harvested and levels of the proteins of interest were identified by Western blotting and Realtime PCR. Distribution of these proteins in the uterus was observed by immunofluorescence. Levels of uterine cAMP were measured by enzyme-linked immunoassay (EIA). Administration of quercetin at increasing doses increased uterine fluid secretion rate, Na+, Cl- and HCO3- concentrations, but to the levels lesser than that of E. In concordant, levels of CFTR, SLC4A4, ENaC (α, β and γ), Na+/K+-ATPase, GPα/β, AC and cAMP in the uterus increased following increased in the doses of quercetin. Co-administration of quercetin with E caused uterine fluid secretion rate, Na+, Cl- and HCO3- concentrations to decrease. In concordant, uterine CFTR, SLC26A6, SLC4A4, ENaC (α, β and γ), Na+/K+-ATPase, GPα/β, AC and cAMP decreased. Greatest effects were observed following co-administration of 10mg/kg/day quercetin with E. Co-administration of quercetin with E+P caused uterine fluid Na+ and HCO3- concentrations to increase but no changes in fluid secretion rate and Cl- concentration were observed. Co-administration of high dose quercetin (100 mg/kg/day) with E+P caused uterine CFTR, SLC26A6, AC, GPα/β and ENaC (α, β and γ) to increase. Quercetin-induced changes in the uterine fluid secretion rate and electrolytes concentrations could potentially affect the uterine reproductive functions under female sex-steroid influence.
子宫内液体环境失调可能会损害成功受孕，这可能是环境雌激素作用的结果。因此，在本研究中，我们在性类固醇的影响下，研究了环境雌激素槲皮素对子宫液体和电解质浓度的影响。对成年雌性去卵巢Sprague-Dawley大鼠皮下注射10、50或100mg/kg/天的槲皮素，并联合17-β雌二醇（E），持续7天；或先注射3天E，然后注射3天E加孕酮（P）（E+P）进行处理。通过体内灌注法测定子宫液体分泌率、Na⁺、Cl⁻和HCO₃⁻浓度。处死后，采集子宫，通过蛋白质免疫印迹法和实时定量PCR鉴定相关蛋白水平。通过免疫荧光观察这些蛋白在子宫内的分布。采用酶联免疫吸附测定（EIA）法测量子宫内cAMP水平。随着槲皮素剂量的增加，子宫液体分泌率、Na⁺、Cl⁻和HCO₃⁻浓度升高，但低于E处理组。同样，随着槲皮素剂量增加，子宫内囊性纤维化跨膜传导调节因子（CFTR）、溶质载体家族4成员4（SLC4A4）、上皮钠通道（ENaC，α、β和γ亚基）、钠钾ATP酶、鸟苷酸结合蛋白α/β亚基（GPα/β）、腺苷酸环化酶（AC）和cAMP水平升高。槲皮素与E联合给药导致子宫液体分泌率、Na⁺、Cl⁻和HCO₃⁻浓度降低。同样，子宫内CFTR、溶质载体家族26成员6（SLC26A6）、SLC4A4、ENaC（α、β和γ亚基）、钠钾ATP酶、GPα/β、AC和cAMP水平降低。槲皮素与E联合给药，剂量为10mg/kg/天时影响最大。槲皮素与E+P联合给药导致子宫液体Na⁺和HCO₃⁻浓度升高，但未观察到液体分泌率和Cl⁻浓度的变化。高剂量槲皮素（100mg/kg/天）与E+P联合给药导致子宫内CFTR、SLC26A6、AC、GPα/β和ENaC（α、β和γ亚基）水平升高。在雌性性类固醇的影响下，槲皮素引起的子宫液体分泌率和电解质浓度变化可能会潜在地影响子宫的生殖功能。
