Abstract：
OBJECTIVE: Aromatase plays an important role in ovarian development, the normal progress of the menstrual cycle, and fertility status. Elevated aromatase activity is linked to obesity. There is a bidirectional relationship between obesity and thyroid function. Few studies have investigated the relationship between TSH and ovarian aromatase in obesity. Our aim was to investigate the effect of TSH on aromatase expression of ovarian granulosa cells in obese mice.
METHODS: Female mice pups were divided into an obesity group and a control group. Obese parameters and the time of pubertal onset were recorded. At the age of 5 weeks, blood and tissues were obtained. Serum aromatase and hormone concentrations were measured using ELISA. The granulosa cells were isolated and exposed to variable concentrations (0 μM, 1 μM, 10 μM, 100 μM) of TSH. The expression of CYP19A1 mRNA and protein were assessed via RT-qPCR and western blot.
RESULTS: In female mice, body weight, Lee\'s obesity index, and serum levels of E2, aromatase, and TSH were significantly higher in the obesity group compared to the control group, whereas the time of pubertal onset and serum T3 and T4 concentrations were significantly lower (all P < 0.001). In granulosa cells, the expression of CYP19A1 mRNA in the obesity group was lower than that in the control group at 1 μM and 100 μM concentrations of TSH (both P < 0.001). The expression of CYP19A1 protein in the obesity group was higher than that in the control group after TSH stimulation (P = 0.014, P < 0.001, and P = 0.004, respectively). With the increase of TSH concentrations, the expression of CYP19A1 mRNA and protein in the two groups significantly increased (all P < 0.001).
CONCLUSIONS: Early puberty and elevated serum aromatase and TSH levels were found in obese female mice. In the granulosa cells of obese mice, TSH directly regulates aromatase expression in a dose-dependent manner.
METHODS: Female mice pups were divided into an obesity group and a control group. Obese parameters and the time of pubertal onset were recorded. At the age of 5 weeks, blood and tissues were obtained. Serum aromatase and hormone concentrations were measured using ELISA. The granulosa cells were isolated and exposed to variable concentrations (0 μM, 1 μM, 10 μM, 100 μM) of TSH. The expression of CYP19A1 mRNA and protein were assessed via RT-qPCR and western blot.
RESULTS: In female mice, body weight, Lee\'s obesity index, and serum levels of E2, aromatase, and TSH were significantly higher in the obesity group compared to the control group, whereas the time of pubertal onset and serum T3 and T4 concentrations were significantly lower (all P < 0.001). In granulosa cells, the expression of CYP19A1 mRNA in the obesity group was lower than that in the control group at 1 μM and 100 μM concentrations of TSH (both P < 0.001). The expression of CYP19A1 protein in the obesity group was higher than that in the control group after TSH stimulation (P = 0.014, P < 0.001, and P = 0.004, respectively). With the increase of TSH concentrations, the expression of CYP19A1 mRNA and protein in the two groups significantly increased (all P < 0.001).
CONCLUSIONS: Early puberty and elevated serum aromatase and TSH levels were found in obese female mice. In the granulosa cells of obese mice, TSH directly regulates aromatase expression in a dose-dependent manner.
摘要：
目的：芳香化酶在卵巢发育中起重要作用，月经周期的正常进展，和生育状况。芳香酶活性升高与肥胖有关。肥胖与甲状腺功能之间存在双向关系。很少有研究探讨TSH与肥胖患者卵巢芳香化酶的关系。我们的目的是研究TSH对肥胖小鼠卵巢颗粒细胞芳香化酶表达的影响。
方法：将雌性小鼠分为肥胖组和对照组。记录肥胖参数和青春期发作时间。在5周龄时，获得血液和组织。使用ELISA测量血清芳香化酶和激素浓度。分离颗粒细胞并暴露于可变浓度（0μM，1μM,10μM,100μM)的TSH。通过RT-qPCR和Westernblot评估CYP19A1mRNA和蛋白的表达。
结果：在雌性小鼠中，体重,Lee的肥胖指数,血清E2、芳香化酶、与对照组相比，肥胖组的TSH明显更高，而青春期发病时间和血清T3和T4浓度显著降低(均P<0.001)。在颗粒细胞中，在TSH浓度为1μM和100μM时，肥胖组CYP19A1mRNA的表达低于对照组（均P<0.001）。TSH刺激后肥胖组CYP19A1蛋白表达高于对照组(P=0.014,P<0.001,P=0.004)。随着TSH浓度的增加,两组CYP19A1mRNA和蛋白表达均显著升高(均P<0.001)。
结论：在肥胖的雌性小鼠中发现青春期早期和血清芳香化酶和TSH水平升高。在肥胖小鼠的颗粒细胞中，TSH以剂量依赖性方式直接调节芳香化酶表达。
方法：将雌性小鼠分为肥胖组和对照组。记录肥胖参数和青春期发作时间。在5周龄时，获得血液和组织。使用ELISA测量血清芳香化酶和激素浓度。分离颗粒细胞并暴露于可变浓度（0μM，1μM,10μM,100μM)的TSH。通过RT-qPCR和Westernblot评估CYP19A1mRNA和蛋白的表达。
结果：在雌性小鼠中，体重,Lee的肥胖指数,血清E2、芳香化酶、与对照组相比，肥胖组的TSH明显更高，而青春期发病时间和血清T3和T4浓度显著降低(均P<0.001)。在颗粒细胞中，在TSH浓度为1μM和100μM时，肥胖组CYP19A1mRNA的表达低于对照组（均P<0.001）。TSH刺激后肥胖组CYP19A1蛋白表达高于对照组(P=0.014,P<0.001,P=0.004)。随着TSH浓度的增加,两组CYP19A1mRNA和蛋白表达均显著升高(均P<0.001)。
结论：在肥胖的雌性小鼠中发现青春期早期和血清芳香化酶和TSH水平升高。在肥胖小鼠的颗粒细胞中，TSH以剂量依赖性方式直接调节芳香化酶表达。
