GPER-ERK通路在DES影响小鼠睾丸引带发育中的作用

发布时间：2019-04-15 11:57

【摘要】：目的:检测G蛋白偶联雌激素受体(GPER)在小鼠睾丸引带上的存在情况,并进一步探讨已烯雌酚(DES)影响小鼠睾丸引带细胞的非基因效应,从睾丸外因素初步了解外源性雌激素影响雄性生殖系统的可能机制。方法:应用免疫组织化学方法了解GPER在不同发育时期(孕17、19 d,生后0、3、7、14和21 d)小鼠睾丸引带上的表达分布情况,并利用免疫电镜进行亚细胞水平的定位。将培养传代的小鼠睾丸引带细胞随机分为3组:实验组(DES组),经典雌激素核受体阻断剂亦是GPER激动剂ICI 182780干预组(ICI 182780-DES组)和GPER阻断剂G15干预组(G15-DES组)。利用Western Blot方法检测不同组别中DES对其ERK通路的影响。结果:GPER存在于不同发育时期的小鼠睾丸引带内层疏松间叶组织区,主要表达于引带细胞的细胞膜及细胞浆上;免疫电镜显示GPER颗粒主要锚定在小鼠睾丸引带细胞的粗面内质网上,表达量少。Western blot方法发现DES对ERK通路上p-ERK1/2的抑制作用可被GPER阻断剂G15阻断,但预先加入ER阻断剂ICI 182780后没有得到类似的反转现象。结论:GPER持续存在小鼠睾丸引带的各个发育阶段,表达于引带细胞的膜性结构上;DES对小鼠睾丸引带的快速非基因效应至少部分是通过GPER介导的。以睾丸引带为研究对象,探讨DES影响雄性生殖系统发育的可能机制,可为其他迫切需要进行效应评价研究的环境雌激素提供一个新的实验参考系统。
[Abstract]:Objective: to detect the presence of G-protein-coupled estrogen receptor (GPER) in mouse testicular banding, and to further explore the non-genetic effect of diethylstilbestrol (DES) on mouse testicular banding cells. The possible mechanism of the effect of exogenous estrogen on the male reproductive system was preliminarily understood from the extratesticular factors. Methods: immunohistochemical method was used to study the expression and distribution of GPER in testis of mice at different developmental stages (17, 19 days, 0, 3, 7, 14 and 21 days after birth), and the localization of subcellular level was carried out by immunoelectron microscope (IEM) in different stages of development (17 and 19 days of gestation, 0, 3, 7, 14 and 21 days after birth). Three groups were randomly divided into three groups: experimental group (DES group), classical estrogen receptor blocker ICI 182780 intervention group (ICI 182780-DES group) and GPER blocker G15 intervention group (G15-DES group). Western Blot method was used to detect the effect of DES on ERK pathway in different groups. Results: GPER was present in the loose interlobar tissue in the inner layer of the testis of mice at different developmental stages, and was mainly expressed on the cell membrane and cytoplasm of the striated cells. Immunoelectron microscopy showed that GPER granules were mainly anchored in the rough endoplasmic reticulum of mouse testicular lead cells, and the inhibitory effect of DES on p-ERK1/2 in ERK pathway could be blocked by GPER blocker G15 by the method of less. Western blot expression. However, there was no similar reversal phenomenon after adding ER blocker ICI 182780 in advance. Conclusion: GPER continues to exist at all developmental stages of mouse testicular banding, which is expressed in the membrane structure of mouse testicular banding cells, and the rapid non-gene effect of DES on mouse testicular banding is mediated at least in part by GPER. In order to explore the possible mechanism of DES affecting the development of male reproductive system, a new experimental reference system was provided for other environmental estrogens which urgently need to evaluate the effect of testicular crest.
【作者单位】：汕头大学医学院第二附属医院小儿外科;深圳大学附属妇女儿童医院小儿外科;汕头大学医学院第二附属医院儿科;
【基金】：国家自然科学基金(81341099,81170555)
【分类号】：R339.21

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