本文選題：犬 + 子宮內(nèi)膜基質(zhì)細(xì)胞　； 參考：《天津農(nóng)學(xué)院》2015年碩士論文

【摘要】：犬子宮蓄積類疾病多由子宮內(nèi)膜功能異常引起，而子宮內(nèi)膜的調(diào)控又與性激素受體數(shù)量密切相關(guān)。目前，不同類固醇激素對犬子宮內(nèi)膜的作用機(jī)理尚不清楚，因此，本研究探索了在體外培養(yǎng)的犬子宮內(nèi)膜基質(zhì)細(xì)胞（ESCs）中，類固醇激素對PR（孕酮受體）和ERβ（雌激素受體β）的表達(dá)調(diào)控作用。 通過分離子宮蓄膿犬與正常犬ESCs，觀察其生長特性并對其進(jìn)行細(xì)胞免疫熒光鑒定，分析比較該細(xì)胞在病犬中是否異常。結(jié)果顯示，輕度子宮蓄膿犬和正常犬ESCs形態(tài)和特性基本一致，前者的細(xì)胞活性更強(qiáng)，增殖速度較快，可在接種后4天融合成片。經(jīng)細(xì)胞免疫熒光鑒定，該細(xì)胞表達(dá)波形蛋白，陽性率達(dá)98%以上，由此表明本試驗(yàn)所分離的細(xì)胞為犬子宮內(nèi)膜基質(zhì)細(xì)胞。 在試驗(yàn)中，將72孔子宮內(nèi)膜基質(zhì)細(xì)胞隨機(jī)分成8組，每組9孔，每組細(xì)胞都用不同的類固醇激素（17β雌二醇、孕酮、米非司酮）進(jìn)行處理。在含激素培養(yǎng)基中維持培養(yǎng)48h后，，通過qPCR法和免疫熒光法對細(xì)胞中的PR和ERβ進(jìn)行檢測，得到的結(jié)果基本一致。結(jié)果顯示，雌二醇能上調(diào)子宮內(nèi)膜基質(zhì)細(xì)胞中PR和ERβ的表達(dá)，孕酮能下調(diào)PR和ERβ的表達(dá)，雌二醇和孕酮聯(lián)用時(shí)則使PR和ERβ的表達(dá)量達(dá)到最高。米非司酮能增加PR的表達(dá)，減少ERβ的表達(dá)，其同雌二醇、孕酮聯(lián)合應(yīng)用時(shí)，能維持PR和ERβ基本處于正常水平。 本研究建立了犬子宮內(nèi)膜基質(zhì)細(xì)胞的體外細(xì)胞模型，并檢測了類固醇激素對該細(xì)胞中PR和ERβ表達(dá)量的影響。該成果不但可以減少相關(guān)研究中的活體實(shí)驗(yàn)，還為闡明犬子宮蓄積類疾病的發(fā)病機(jī)制提供了一定的理論基礎(chǔ)。
[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.
【學(xué)位授予單位】：天津農(nóng)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.292

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