类固醇激素对犬子宫内膜基质细胞中PR和ERβ表达量的影响

发布时间：2018-06-28 07:32

本文选题：犬 + 子宫内膜基质细胞　；参考：《天津农学院》2015年硕士论文

【摘要】：犬子宫蓄积类疾病多由子宫内膜功能异常引起，而子宫内膜的调控又与性激素受体数量密切相关。目前，不同类固醇激素对犬子宫内膜的作用机理尚不清楚，因此，本研究探索了在体外培养的犬子宫内膜基质细胞（ESCs）中，类固醇激素对PR（孕酮受体）和ERβ（雌激素受体β）的表达调控作用。通过分离子宫蓄脓犬与正常犬ESCs，观察其生长特性并对其进行细胞免疫荧光鉴定，分析比较该细胞在病犬中是否异常。结果显示，轻度子宫蓄脓犬和正常犬ESCs形态和特性基本一致，前者的细胞活性更强，增殖速度较快，可在接种后4天融合成片。经细胞免疫荧光鉴定，该细胞表达波形蛋白，阳性率达98%以上，由此表明本试验所分离的细胞为犬子宫内膜基质细胞。在试验中，将72孔子宫内膜基质细胞随机分成8组，每组9孔，每组细胞都用不同的类固醇激素（17β雌二醇、孕酮、米非司酮）进行处理。在含激素培养基中维持培养48h后，，通过qPCR法和免疫荧光法对细胞中的PR和ERβ进行检测，得到的结果基本一致。结果显示，雌二醇能上调子宫内膜基质细胞中PR和ERβ的表达，孕酮能下调PR和ERβ的表达，雌二醇和孕酮联用时则使PR和ERβ的表达量达到最高。米非司酮能增加PR的表达，减少ERβ的表达，其同雌二醇、孕酮联合应用时，能维持PR和ERβ基本处于正常水平。本研究建立了犬子宫内膜基质细胞的体外细胞模型，并检测了类固醇激素对该细胞中PR和ERβ表达量的影响。该成果不但可以减少相关研究中的活体实验，还为阐明犬子宫蓄积类疾病的发病机制提供了一定的理论基础。
[Abstract]:The accumulation diseases in dogs are mostly caused by abnormal endometrial function, and the regulation of endometrium is closely related to the number of sex hormone receptors. At present, the mechanism of different steroid hormones on canine endometrium is unclear. Therefore, in vitro cultured canine endometrial stromal cells (ESCs) were investigated in this study. Steroid hormones regulate the expression of PR (progesterone receptor) and ER 尾 (estrogen receptor 尾). By separating suppurative canine and normal dog ESCs, the growth characteristics of ESCs were observed, and the cell immunofluorescence was used to analyze and compare whether the cells were abnormal in the diseased dogs. The results showed that the morphology and characteristics of ESCs in mild purulent dogs and normal dogs were basically the same. The former had stronger cell activity and faster proliferation, and could be fused into slices 4 days after inoculation. The positive rate of vimentin was over 98%, which indicated that the cells isolated in this experiment were canine endometrial stromal cells. In the experiment, 72 well endometrial stromal cells were randomly divided into 8 groups with 9 holes in each group. Each group was treated with different steroid hormones (17 尾 -estradiol, progesterone, mifepristone). After 48 hours of culture in steroid medium, PR and ER 尾 were detected by qPCR and immunofluorescence, and the results were basically the same. The results showed that estradiol could up-regulate the expression of PR and ER 尾 in endometrial stromal cells, progesterone could down-regulate the expression of PR and ER 尾, and the expression of PR and ER 尾 was the highest when combined with estradiol and progesterone. Mifepristone could increase the expression of PR and decrease the expression of ER 尾. When combined with estradiol and progesterone, mifepristone could maintain PR and ER 尾 at normal level. In this study, a canine endometrial stromal cell model was established in vitro, and the effects of steroid hormones on the expression of PR and ER 尾 were detected. The results can not only reduce the living experiments in related studies, but also provide a theoretical basis for elucidating the pathogenesis of canine uterine accumulative diseases.
【学位授予单位】：天津农学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.292

【参考文献】