BDE-209對(duì)小鼠卵母細(xì)胞體外成熟和孤雌激活后孤雌胚發(fā)育的影響
發(fā)布時(shí)間:2018-02-25 00:12
本文關(guān)鍵詞: 十溴聯(lián)苯醚 小鼠 卵母細(xì)胞 體外成熟 體外培養(yǎng) 孤雌生殖 出處:《暨南大學(xué)》2009年碩士論文 論文類型:學(xué)位論文
【摘要】: 目的: 研究十溴聯(lián)苯醚(brominated diphenyl ethers-209,BDE-209)對(duì)小鼠卵母細(xì)胞體外成熟過程中生發(fā)泡破裂率、第一極體釋放率和卵母細(xì)胞存活率的影響,并對(duì)培養(yǎng)至成熟卵母細(xì)胞行體外孤雌激活處理,研究孤雌激活后BDE-209對(duì)胚胎發(fā)育的影響。 方法: 通過對(duì)性成熟未孕健康母鼠進(jìn)行PMSG+HCG超排卵處理,取得成熟卵母細(xì)胞。將0μg/ml BDE-209作為對(duì)照組A,10、20、40μg/ml作為實(shí)驗(yàn)組B、C、D,用含A、B、C、D不同濃度BDE-209的DMEM(低糖)培養(yǎng)液行體外培養(yǎng),觀察各組在8小時(shí)和16小時(shí)的第一極體釋放率和卵母細(xì)胞存活率,并比較其差異。用PMSG促排卵處理從性未成熟小鼠體內(nèi)獲得未成熟卵母細(xì)胞行體外培養(yǎng),于12小時(shí)觀察卵母細(xì)胞生發(fā)泡破裂率,于24小時(shí)觀察第一極體釋放率。再將MⅡ期卵母細(xì)胞行乙醇孤雌激活處理,比較各組孤雌激活率是否有差異,將孤雌激活后的卵母細(xì)胞置于含不同濃度BDE-209的CZB培養(yǎng)液中行胚胎培養(yǎng),于培養(yǎng)的24小時(shí)觀察2-細(xì)胞形成率,于48小時(shí)觀察4-細(xì)胞形成率,并比較其組間差異。 結(jié)果: 對(duì)成熟卵母細(xì)胞行體外培養(yǎng),分別比較A、B、C、D各組8小時(shí)和16小時(shí)第一極體釋放率,各組差別無統(tǒng)計(jì)學(xué)意義(P>0.05)。于8小時(shí)和16小時(shí)兩時(shí)段分別比較四組間其第一極體釋放率差別均無統(tǒng)計(jì)學(xué)意義(P>0.05);分別比較A、B、C、D各組8小時(shí)和16小時(shí)的卵母細(xì)胞存活率,差別無統(tǒng)計(jì)學(xué)意義(P>0.05)。B、C、D各組16小時(shí)組比8小時(shí)組卵母細(xì)胞存活率均有所降低,P<0.05,差別統(tǒng)計(jì)學(xué)意義。于8小時(shí),16小時(shí)時(shí)段分別比較四組間卵母細(xì)胞存活率,P>0.05,差別無統(tǒng)計(jì)學(xué)意義。 對(duì)未成熟卵母細(xì)胞行體外培養(yǎng),比較各組12小時(shí)生發(fā)泡破裂率,P>0.05,差別無統(tǒng)計(jì)學(xué)意義。比較24小時(shí)各組第一極體釋放率,實(shí)驗(yàn)組比對(duì)照組均有明顯降低,P<0.05,差別有統(tǒng)計(jì)學(xué)意義。將各組培養(yǎng)所得成熟卵母細(xì)胞行孤雌激活處理,P>0.05,各組的激活率無明顯差異。 將孤雌激活后的卵母細(xì)胞行體外胚胎培養(yǎng),于24小時(shí)觀察2-細(xì)胞形成率,48小時(shí)觀察4-細(xì)胞形成率,實(shí)驗(yàn)組較對(duì)照組有所降低,P<0.05,差別有統(tǒng)計(jì)學(xué)意義。 結(jié)論: BDE-209對(duì)成熟卵母細(xì)胞的培養(yǎng)過程中,不抑制小鼠卵母細(xì)胞第一極體釋放率,但降低卵母細(xì)胞的存活率。對(duì)未成熟卵母細(xì)胞的體外培養(yǎng)過程中,其不抑制生發(fā)泡破裂,但是使得第一極體釋放率降低。對(duì)培養(yǎng)成熟的卵母細(xì)胞行孤雌激活處理,各組間激活率無顯著性差異,但其對(duì)孤雌激活后胚胎發(fā)育有明顯的抑制作用。
[Abstract]:Objective:. To study the effects of decabrominated diphenyl ethers-209 (BDE-209) on blastocyst rupture rate, first polar body release rate and oocyte survival rate during mouse oocytes maturation in vitro, and parthenogenetic activation of cultured mature oocytes in vitro. To study the effect of parthenogenetic activated BDE-209 on embryonic development. Methods:. The mature oocytes were obtained by superovulation of PMSG HCG on unpregnant and sexual mature healthy mice. 0 渭 g / ml BDE-209 was used as the control group (AH1010, 2040 渭 g / ml) as the experimental group, and DMEM (low glucose) culture medium containing different concentrations of BDE-209 was cultured in vitro. The first polar body release rate and oocyte survival rate at 8 and 16 hours in each group were observed and the difference was compared. Immature oocytes were obtained from immature mice by PMSG stimulation in vitro. The blastocyst rupture rate of oocytes was observed at 12 hours and the release rate of the first polar body was observed at 24 hours. Then the M 鈪,
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