本文選題：子宮液　切入點(diǎn)：外泌體　出處：《西北農(nóng)林科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：外泌體是由細(xì)胞通過(guò)胞吞胞吐的方式向外環(huán)境中釋放的一種納米級(jí)囊泡,其內(nèi)包含重要的生物信息物質(zhì),如蛋白質(zhì)、脂質(zhì)、mRNA和mi RNA等。這種小囊泡作為“信使”廣泛參與著機(jī)體生理或病理的生物學(xué)活動(dòng)。越來(lái)越多的研究表明,外泌體幾乎存在于所有的體液當(dāng)中,暗示其對(duì)生物生命活動(dòng)的重要性以及必要性。有證據(jù)表明從哺乳動(dòng)物生命的最開(kāi)始——胚胎,外泌體就扮演了很重要的角色。在本研究中,首先對(duì)發(fā)情期牛的子宮液中的外泌體進(jìn)行提取和鑒定,之后將其添加至IVF培養(yǎng)液中,來(lái)觀察其對(duì)IVF胚胎發(fā)育的影響,主要研究結(jié)果如下:1.利用導(dǎo)管向發(fā)情期的牛子宮注入生理鹽水進(jìn)行沖洗,每次2 ml,重復(fù)6-8次,收集到子宮沖洗液;2.通過(guò)改良后的超速離心法,獲得了純度較高的子宮液外泌體;3.在透射電子顯微鏡觀察到外泌體形態(tài):直徑在100 nm左右,大小均一,形態(tài)較為規(guī)則,呈圓形或橢圓形;4.外泌體特異性膜蛋白免疫熒光染色顯示為CD9+,Western Blot曝光出CD9條帶;5.將外泌體樣品添加至IVF培養(yǎng)液中,結(jié)果顯示實(shí)驗(yàn)(exosomes)組與對(duì)照組的卵裂率(81.2%vs79.1%)以及囊胚率(33.3%vs32.5%)均未見(jiàn)明顯差異;6.經(jīng)囊胚的凋亡染色,在囊胚細(xì)胞數(shù)(110.21±4.52 vs 113.11±3.20)無(wú)差異的情況下,實(shí)驗(yàn)(exosome)組囊胚凋亡細(xì)胞數(shù)顯著低于對(duì)照組(2.68±0.30 vs 5.83±0.44,P0.05)。綜上所述,發(fā)情期的牛子宮液外泌體能夠降低體外培養(yǎng)囊胚細(xì)胞凋亡的比例。
[Abstract]:Exosomes are nanometer-sized vesicles released by cells into the outer environment by emesis. They contain important biological information substances, such as proteins. These vesicles, as messengers, are widely involved in the biological activities of the body's physiology or pathology. More and more studies have shown that exocrine bodies are present in almost all body fluids. Suggesting its importance and necessity for biological activities. There is evidence that exosomes play a significant role from the very beginning of mammalian life, the embryo. In order to observe the effect of estrus on the development of IVF embryos, the exocrine in the uterine fluid of estrous cattle was extracted and identified, and then added into the IVF culture medium to observe its effect on the development of IVF embryos. The main results are as follows: 1. Irrigating bovine uterus during estrus with normal saline with catheter, 2 ml each time, repeat 6 to 8 times, collect uterine irrigation fluid 2. Through the improved ultracentrifugation method, The exocrine of uterine fluid with high purity was obtained. The morphology of exocrine was observed by transmission electron microscope (TEM): the diameter was about 100 nm, the size was uniform, and the morphology was regular. The exocrine specific membrane protein immunofluorescence staining showed that the CD9 band was exposed by CD9 Blot. The exocrine sample was added to the culture medium of IVF. The results showed that there was no significant difference in cleavage rate (81.2 vs 79.1) and blastocyst rate (33.3vs32.5) between the experimental group and the control group. There was no significant difference in blastocyst cell number (110.21 鹵4.52 vs 113.11 鹵3.20). The number of blastocyst apoptotic cells in the estrus group was significantly lower than that in the control group (2.68 鹵0.30 vs 5.83 鹵0.44p 0.05). In conclusion, the exocrine of bovine uterus during estrus could reduce the percentage of apoptosis of blastocyst cells cultured in vitro.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S823

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