本文選題：外源性內脂素　切入點：誘導排卵　出處：《河北聯(lián)合大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的通過檢測注射外源性內脂素的高齡雌鼠卵巢組織血管內皮生長因子（vascular endothelial growth factor，VEGF）和內分泌腺性血管內皮生長因子（endocrine gland vascular endothelial growth factor，EG-VEGF）的表達來探討內脂素對高齡雌鼠卵巢功能和卵子質量的影響，為臨床促排卵中卵巢低反應患者的用藥提供實驗依據(jù)。 方法將兩組不同周齡段（12～16周，32～36周）的SPF級雌性SD大鼠每組各60只，每組內再隨機分為兩組。在誘導排卵的基礎上，應用外源性內脂素的作為治療組，將僅誘導排卵的一組作為對照組。第一步，誘導排卵，于發(fā)情期給予四組雌性SD大鼠腹腔內注射15IU孕母馬血清促性腺激素，同時，治療組予100ng/ml重組鼠內脂素。第二步，注射48小時后，向治療組與對照組大鼠腹腔內注射50IU人絨毛膜促性腺激素（HCG）。第三步，立即將這些雌鼠和雄鼠進行單獨交配。第四步，交配18小時后，頸椎脫臼處死雌鼠，無菌條件下取出雙側輸卵管中的受精卵進行體外培養(yǎng)；取出雙側卵巢，右側卵巢用甲醛固定，免疫組化檢測微血管密度（MVD），左側卵巢的一部分立即置于Trizol中，用于Realtime-PCR檢測卵巢VEGF、EG-VEGF的mRNA表達，另一部分則用于Western Blotting檢測VEGF、EG-VEGF的蛋白表達。實驗數(shù)據(jù)采用SPSS17.0軟件進行統(tǒng)計學分析，計量資料以平均數(shù)±標準差（x s）表示，采用單因素方差分析判斷組間差異，兩組間數(shù)據(jù)比較采用SNK檢驗，P0.05認為有統(tǒng)計學意義。 結果1年輕對照組平均合子數(shù)、卵裂數(shù)、卵裂率、平均胚胎數(shù)高于大齡對照組，，差異有統(tǒng)計學意義（P＜0.05），大齡治療組高于大齡對照組，差異有統(tǒng)計學意義（P＜0.05）；而年輕治療組、年輕對照組兩組之間比較，差異無統(tǒng)計學意義（P＞0.05）。2年輕對照組卵巢MVD、VEGF及EG-VEGF的表達水平高于大齡對照組，差異有統(tǒng)計學意義（P＜0.05），大齡治療組高于大齡對照組，差異有統(tǒng)計學意義（P＜0.05）；而年輕治療組、年輕對照組兩組之間比較，差異無統(tǒng)計學意義（P＞0.05）。 結論1誘導排卵的高齡雌鼠的平均合子數(shù)、卵裂數(shù)、卵裂率、平均胚胎數(shù)隨周齡的增加而下降，提示高齡雌鼠生殖能力下降。2給予外源性Qg脂素后治療組高齡雌鼠的平均合子數(shù)、卵裂數(shù)、卵裂率、平均胚胎數(shù)明顯增加，提示外源性Qg脂素可能改善高齡雌鼠卵巢功能，提高其生殖能力。3給予外源性Qg脂素后高齡雌鼠卵巢MVD、VEGF、EG-VEGF的表達水平明顯提高，提示外源性Qg脂素可能是通過促進卵巢血管的生成從而改善高齡雌鼠的卵巢功能和生殖能力。
[Abstract]:Objective to investigate the expression of vascular endothelial growth factor (VEGF) and secreted glandular vascular endothelial growth factor (VEGF) in ovarian tissues of elderly female rats injected with exogenous endolipin, and to explore the effect of endocrine gland vascular endothelial growth factor-EG-VEGFF on the oocytes of elderly female rats. The effects of nest function and egg quality, To provide experimental evidence for the treatment of ovarian hyporeactive patients in clinical ovulation promotion. Methods two groups of female SD rats of SPF grade were randomly divided into two groups. On the basis of ovulation induction, exogenous endolipin was used as treatment group. In the first step, ovulation was induced by intraperitoneal injection of serum gonadotropin in female SD rats of 15 IU during estrus. Meanwhile, the treatment group was given 100ng / ml recombinant rat lipopolipin. After 48 hours of injection, 50 IU human chorionic gonadotropin (HCG) was injected intraperitoneally into the treatment group and the control group. In the third step, the female and male mice were mated separately. 4th steps, 18 hours after mating, the female rats were killed by cervical dislocations. Fertilized eggs from bilateral oviducts were removed for culture in vitro under aseptic conditions, bilateral ovaries were removed, the right ovaries were fixed with formaldehyde, microvessel density was detected by immunohistochemistry, and a portion of the left ovary was placed in Trizol immediately. The Realtime-PCR was used to detect the expression of EG-VEGF in ovary and the other part was used to detect the protein expression of EG-VEGF in VEGFU by Western Blotting. The experimental data were analyzed statistically by SPSS17.0 software, and the measured data were expressed as mean 鹵standard deviation (x / s). Single factor analysis of variance was used to judge the difference between the two groups. The data of the two groups were compared by SNK test (P0.05). Results 1the average number of zygotes, cleavage, cleavage rate and the number of embryos in the young control group were higher than those in the older control group (P < 0.05), the difference was statistically significant (P < 0.05), and the difference was statistically significant in the older treatment group than in the older control group (P < 0.05), while in the young treatment group, the difference was significant (P < 0.05). There was no significant difference between the two groups (P > 0.05). 2 the expression of EG-VEGF and VEGF in the ovary of the young control group was higher than that of the older control group (P < 0.05), and the expression of EG-VEGF in the older group was higher than that in the older control group (P < 0.05). The difference was statistically significant (P < 0.05), but there was no significant difference between the young treatment group and the young control group (P > 0.05). Conclusion 1 the average number of zygotes, cleavage, cleavage rate and the average number of embryos decreased with the increase of week age. It was suggested that the average number of zygotes, cleavage, cleavage rate and the average number of embryos were significantly increased in the treatment group, which suggested that exogenous QG lipopolipin might improve the ovarian function of the aged female rats. After treated with exogenous QG lipopolipin, the expression of EG-VEGF in ovaries of aged female rats was significantly increased, suggesting that exogenous QG lipopolipin might improve the ovarian function and reproductive ability of elderly female rats by promoting the formation of ovarian blood vessels.
【學位授予單位】：河北聯(lián)合大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R714.8

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