本文選題：支持細胞 + FSH��； 參考：《東北農(nóng)業(yè)大學》2015年碩士論文

【摘要】：睪丸支持細胞位于曲精細管中,是精子發(fā)生的微環(huán)境的主要構(gòu)成。許多激素和影響因子都參與了睪丸支持細胞增殖調(diào)控,其中FSH是調(diào)節(jié)睪丸支持細胞增殖進程的重要激素之一。因此,研究FSH對犢牛睪丸支持細胞增殖的調(diào)節(jié)機制不僅能為人工調(diào)控支持細胞增殖奠定一定基礎(chǔ),而且對提高雄性動物的生殖能力也具有十分重要的意義。本實驗以新生犢牛睪丸為實驗材料,利用胰酶、膠原酶兩步消化法對支持細胞進行分離,差速貼壁法純化支持細胞,進行孚爾根染色法、HE染色法以及油紅O染色法鑒定支持細胞;以培養(yǎng)的犢牛支持細胞為實驗材料,用不同濃度的FSH(0、0.01、0.02、0.04、0.08 IU/ml)對支持細胞進行處理,利用CCK8檢測五組濃度6個時間點支持細胞增殖情況并用RealtimePCR檢測一定濃度FSH處理后CDC25等基因表達情況。兩步酶法可以對犢牛睪丸支持細胞進行分離純化,經(jīng)孚爾根染色等方法鑒定純度大于95%;體外培養(yǎng)條件下的支持細胞能夠傳至第20代,并且隨著傳代次數(shù)的增加,支持細胞的增殖速度越來越慢。FSH可以顯著提高睪丸支持細胞體外增殖的效率,0.04 IU/ml的FSH能顯著地促進支持細胞體外增殖。FSH顯著促進WNT/β-catenin信號通路相關(guān)基因表達量的提高,可以通過該信號通路促進支持細胞的增殖。FSH對CDC25A基因表達影響不顯著,在一定濃度范圍內(nèi),FSH顯著促進CDC25B的表達。CDC25B可能參與促進支持細胞增殖作用,其可能決定支持細胞的增殖速度。在一定濃度范圍內(nèi),FSH顯著促進P53的表達量,P53可能在FSH刺激性下對支持增殖起到控制作用;FSH顯著抑制FAS和CDC25C的表達量,同時顯著提高FASL的表達量。
[Abstract]:Testicular Sertoli cells, located in the tubules, are the main components of the microenvironment of spermatogenesis. Many hormones and influencing factors are involved in the regulation of testicular Sertoli cell proliferation, and FSH is one of the important hormones to regulate the proliferation of testicular Sertoli cells. Therefore, the study of the regulatory mechanism of FSH on the proliferation of testicular Sertoli cells in calves can not only lay a foundation for artificial regulation of Sertoli cell proliferation, but also play an important role in improving the reproductive ability of male animals. Sertoli cells were isolated from newborn calf testis by trypsin and collagenase digestion, and Sertoli cells were purified by differential adhesion method. Sertoli cells were identified by HE staining and oil red O staining, and Sertoli cells were treated with different concentrations of FSHO 0. 01 and 0. 02O 0. 04U / ml, respectively, and cultured calf Sertoli cells were used as experimental materials. CCK8 was used to detect the proliferation of Sertoli cells at 6 time points in five groups and RealtimePCR was used to detect the expression of CDC25 and other genes after treatment with certain concentration of FSH. Two-step enzymatic method was used to isolate and purify testicular Sertoli cells from calves. The purity of Sertoli cells in vitro was higher than 95. The Sertoli cells cultured in vitro could be transmitted to the 20th passage, and with the increase of passage times, the purity of Sertoli cells was higher than 95%. The proliferation of Sertoli cells was slower and slower. FSH could significantly improve the proliferation efficiency of testicular Sertoli cells in vitro. FSH of 0.04 IU/ml could significantly promote the proliferation of Sertoli cells in vitro. FSH significantly promoted the expression of genes related to WNT/ 尾 -catenin signaling pathway. FSH can promote the proliferation of Sertoli cells through this signaling pathway. FSH has no significant effect on the expression of CDC25A gene. In a certain concentration range, FSH can significantly promote the expression of CDC25B. CDC25B may play a role in promoting the proliferation of Sertoli cells. It may determine the rate of proliferation of Sertoli cells. In a certain concentration range, FSH significantly promoted the expression of p53. The expression of p53 may play a controlling role in supporting proliferation under the stimulation of FSH. FSH significantly inhibited the expression of FAS and CDC25C, and significantly increased the expression of FASL.
【學位授予單位】：東北農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S814

【參考文獻】

相關(guān)期刊論文 前2條

1 Li Yu-long;Wu Qiong;Zhao Xun-wu;Zeng Yue;Elkanah Adegoke;Zhang Gui-xue;;Influence of FSH Treatment on Expression of CDC25A, TSSK3 and P53 in Vitro Cultured Sertoli Cells of Calf[J];Journal of Northeast Agricultural University(English Edition);2015年01期

2 時宇;于娜;劉欣;AHMED KHALID;林旭;李玉龍;張貴學;;犢牛睪丸支持細胞的培養(yǎng)及不同濃度pEGFP-N3載體瞬時轉(zhuǎn)染[J];黑龍江畜牧獸醫(yī);2014年13期

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本文編號：1928388