EGCG對水牛卵母細胞體外成熟和體外受精的影響
發(fā)布時間：2018-07-17 16:58
【摘要】：體外胚胎生產(chǎn)中,各種抗氧化劑常被添加于不同的培養(yǎng)液中以提高體外胚胎生產(chǎn)效率。實驗表明,表沒食子兒茶素沒食子酸酯(Epigallocatechin Gallate, EGCG)能夠清除健康細胞中ROS,提高配子的抗氧化能力,有利于動物卵母細胞體外成熟和精子受精。本研究主要探討了EGCG對水牛卵子體外成熟、精子受精能力的影響以及可能的作用機理,旨在使水牛體外胚胎生產(chǎn)高效化。本研究包括三個試驗：試驗一,EGCG對水牛卵母細胞體外成熟的影響。旨在探討EGCG對水牛卵母細胞卵丘擴展、成熟率的影響,并初步試驗EGCG對水牛配子發(fā)育潛能的影響。結(jié)果發(fā)現(xiàn)：(1)10、20 μmol/L處理組與對照組卵丘擴展指數(shù)差異顯著(2.49、2.42 vs.2.22, P0.05);5、30 μmol/L處理組卵丘擴展指數(shù)也高于對照組,但無統(tǒng)計學(xué)意義(2.36、2.28 vs.2.22,P0.05)；(2)10、20μmol/L處理組成熟率顯著高于對照組(59.31%、50.98% vs.45.26%,P0.05)；但兩組之間差異不顯著；5 μmol/L和30 μmol/L處理組的成熟率與對照組差異不顯著(47.04%、42.81% vs.45.26%,P0.05)；(3)在成熟液中添加EGCG,各處理組的卵裂率、囊胚率均高于對照組,其中10 μmol/L處理組效果最佳。孤雌激活后,卵裂率和囊胚率均顯著高于對照組(67.07%vs.57.53%、31.69% vs.20.66%, p0.05);體外受精后,卵裂率和囊胚率也顯著高于對照組(59.64% vs.44.65%、24.68% vs.15.02%, p0.05)。以上結(jié)果表明,EGCG能促進水牛卵母細胞體外成熟,并提高了卵母細胞的發(fā)育潛能。試驗二,EGCG (0、5、10.20、30 μmol/L)對水牛卵母細胞體外成熟階段抗氧化能力的影響。結(jié)果顯示：(1)添加20.30 μmol/L的處理組MⅡ期ROS DCF熒光強度與對照組差異顯著(18.77、25.39 vs.38.04,P0.05)；(2)各處理組MⅡ期水牛卵母細胞H202含量與對照組差異不顯著(4.68、4.33、4.16、4.51 vs. 4.90, P0.05); (3) 5、10、20 μmol/L處理組GSH含量與對照組差異顯著(3.0、3.21、3.25 vs.2.59, P0.05); 30 μmol/L處理組GSH含量也高于對照組,但無統(tǒng)計學(xué)意義(2.99 vs.2.59,P0.05)。以上結(jié)果表明：在水牛卵母細胞體外成熟過程中,EGCG能夠降低卵母細胞內(nèi)活性氧水平,提高GSH的合成,但不能降低H202的含量；(4)綜上所述,在卵母細胞體外成熟過程中,EGCG影響氧化反應(yīng)。試驗三,探討在受精液中添加10 μmol/L的EGCG對水牛精子受精能力的影響以及在成熟液和受精液中同時添加EGCG是否有協(xié)同作用。結(jié)果顯示：在成熟液、受精液、成熟液和受精液中分別添加10μmol/L的EGCG后,各組卵裂率顯著高于對照組(61.31%、59.14%、58.57% vs.45.71%,P0.05)；各組的囊胚率顯著高于對照組(23.61%%、22.77%、23.56% vs.16.74%,P0.05)。以上結(jié)果表明,EGCG能提高水牛精子受精能力和促進早期胚胎發(fā)育,但在成熟液和受精液中同時添加EGCG沒有協(xié)同作用。為了高效方便,本試驗中,最佳方案為在成熟液或者受精液中添加10 μmol/L的EGCG�？傊�,EGCG作為一種高效的抗氧化劑,能夠促進水牛卵母細胞體外成熟、體外受精,并提高了胚胎體外發(fā)育率。EGCG對提高水牛體外胚胎生產(chǎn)有積極影響,可提高水牛卵子的利用率。本實驗室條件中,單獨在成熟液或者受精液中添加10 μmol/L EGCG效果最好。
[Abstract]:In vitro embryo production, all kinds of antioxidants are often added to different cultures to improve the efficiency of in vitro embryo production. The experiment shows that epigallocatechin gallate (Epigallocatechin Gallate, EGCG) can remove ROS in healthy cells, improve the anti oxygen ability of gametes, and be beneficial to the maturation and sperm of animal oocytes in vitro. This study mainly discussed the effect of EGCG on the maturation of buffalo ovum in vitro, the effect of sperm fertilization and the possible mechanism of action to make the production of buffalo in vitro embryo efficient. This study includes three experiments: Experiment 1, the effect of EGCG on the maturation of buffalo oocytes in vitro. The purpose of this study is to explore the cumulus expansion of the oocytes of the buffalo oocytes. The effect of EGCG on the development potential of buffalo gamete was preliminarily tested. The results were as follows: (1) the cumulus expansion index of 10,20 mol/L treatment group was significantly different from that of control group (2.49,2.42 vs.2.22, P0.05), and the cumulus expansion index in 5,30 mu mol/L treatment group was also higher than that in the control group, but there was no statistical significance (2.36,2.28 vs.2.22, P0.05); (2) 1 The maturity rate of 0,20 mu mol/L treatment group was significantly higher than that of the control group (59.31%, 50.98% vs.45.26%, P0.05), but there was no significant difference between the two groups. The maturity rate of 5 and 30 micron mol/L treatment groups was not significantly different from the control group (47.04%, 42.81% vs.45.26%, P0.05), and (3) adding EGCG in the mature liquid, the cleavage rate of each treatment group and the blastocyst rate were higher than those of the control group. After the parthenogenetic activation, the cleavage rate and blastocyst rate were significantly higher than those in the control group (67.07%vs.57.53%, 31.69% vs.20.66%, P0.05), and the cleavage rate and blastocyst rate were significantly higher than those of the control group (59.64% vs.44.65%, 24.68% vs.15.02%, P0.05) after in vitro fertilization. The results showed that EGCG could promote buffalo oocyte. The effects of two, EGCG (0,5,10.20,30 mol/L) on the antioxidant capacity of buffalo oocytes in vitro were tested. The results showed that: (1) the difference in the fluorescence intensity of M II ROS DCF in the treatment group with 20.30 micron mol/L was significantly different from that of the control group (18.77,25.39 vs.38.04, P0.05); (2) each treatment group The H202 content of M II buffalo oocyte was not significantly different from that of the control group (4.68,4.33,4.16,4.51 vs. 4.90, P0.05), and (3) the GSH content in the 5,10,20 mu mol/L treatment group was significantly different from the control group (3.0,3.21,3.25 vs.2.59, P0.05), and the 30 mu mol/L treatment group was also higher than the control group, but there was no statistical significance (2.99). The above result table In the process of buffalo oocyte maturation in vitro, EGCG can reduce the level of active oxygen in oocyte and improve the synthesis of GSH, but can not reduce the content of H202. (4) in summary, EGCG affects oxidation reaction in the process of oocyte maturation in vitro. Experiment three, to study the fertilization of sperm in semen by adding 10 mu mol/L of EGCG to sperm fertilization. The results showed that the rate of cleavage in each group was significantly higher than that of the control group (61.31%, 59.14%, 58.57% vs.45.71%, P0.05), and the blastocyst rate in each group was significantly higher than that of the control group (23). The results showed that in the mature solution, the mature solution, the mature solution and the semen were added to the EGCG, the percentage of cleavage was significantly higher than that of the control group (61.31%, 59.14%, 58.57% vs.45.71%, P0.05) in the mature solution, and the mature solution and the semen respectively (23). (23 .61%%, 22.77%, 23.56% vs.16.74%, P0.05). The above results show that EGCG can improve the fertilization ability of buffalo sperm and promote the development of early embryo, but there is no synergy between the addition of EGCG in the mature liquid and the semen. In order to be efficient and convenient, the best solution for this experiment is to add 10 u mol/L EGCG. to the mature solution or the semen, EGCG, EGCG. As a highly effective antioxidant, it can promote the maturation of buffalo oocyte in vitro, in vitro fertilization, and increase the development rate of the embryo in vitro,.EGCG has a positive effect on improving the production of buffalo embryos in vitro, and can improve the utilization of buffalo eggs. In the laboratory conditions, the effect of adding 10 mu mol/L EGCG to the mature solution or the semen is the most effective. OK.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S823.83
本文編號：2130306
[Abstract]:In vitro embryo production, all kinds of antioxidants are often added to different cultures to improve the efficiency of in vitro embryo production. The experiment shows that epigallocatechin gallate (Epigallocatechin Gallate, EGCG) can remove ROS in healthy cells, improve the anti oxygen ability of gametes, and be beneficial to the maturation and sperm of animal oocytes in vitro. This study mainly discussed the effect of EGCG on the maturation of buffalo ovum in vitro, the effect of sperm fertilization and the possible mechanism of action to make the production of buffalo in vitro embryo efficient. This study includes three experiments: Experiment 1, the effect of EGCG on the maturation of buffalo oocytes in vitro. The purpose of this study is to explore the cumulus expansion of the oocytes of the buffalo oocytes. The effect of EGCG on the development potential of buffalo gamete was preliminarily tested. The results were as follows: (1) the cumulus expansion index of 10,20 mol/L treatment group was significantly different from that of control group (2.49,2.42 vs.2.22, P0.05), and the cumulus expansion index in 5,30 mu mol/L treatment group was also higher than that in the control group, but there was no statistical significance (2.36,2.28 vs.2.22, P0.05); (2) 1 The maturity rate of 0,20 mu mol/L treatment group was significantly higher than that of the control group (59.31%, 50.98% vs.45.26%, P0.05), but there was no significant difference between the two groups. The maturity rate of 5 and 30 micron mol/L treatment groups was not significantly different from the control group (47.04%, 42.81% vs.45.26%, P0.05), and (3) adding EGCG in the mature liquid, the cleavage rate of each treatment group and the blastocyst rate were higher than those of the control group. After the parthenogenetic activation, the cleavage rate and blastocyst rate were significantly higher than those in the control group (67.07%vs.57.53%, 31.69% vs.20.66%, P0.05), and the cleavage rate and blastocyst rate were significantly higher than those of the control group (59.64% vs.44.65%, 24.68% vs.15.02%, P0.05) after in vitro fertilization. The results showed that EGCG could promote buffalo oocyte. The effects of two, EGCG (0,5,10.20,30 mol/L) on the antioxidant capacity of buffalo oocytes in vitro were tested. The results showed that: (1) the difference in the fluorescence intensity of M II ROS DCF in the treatment group with 20.30 micron mol/L was significantly different from that of the control group (18.77,25.39 vs.38.04, P0.05); (2) each treatment group The H202 content of M II buffalo oocyte was not significantly different from that of the control group (4.68,4.33,4.16,4.51 vs. 4.90, P0.05), and (3) the GSH content in the 5,10,20 mu mol/L treatment group was significantly different from the control group (3.0,3.21,3.25 vs.2.59, P0.05), and the 30 mu mol/L treatment group was also higher than the control group, but there was no statistical significance (2.99). The above result table In the process of buffalo oocyte maturation in vitro, EGCG can reduce the level of active oxygen in oocyte and improve the synthesis of GSH, but can not reduce the content of H202. (4) in summary, EGCG affects oxidation reaction in the process of oocyte maturation in vitro. Experiment three, to study the fertilization of sperm in semen by adding 10 mu mol/L of EGCG to sperm fertilization. The results showed that the rate of cleavage in each group was significantly higher than that of the control group (61.31%, 59.14%, 58.57% vs.45.71%, P0.05), and the blastocyst rate in each group was significantly higher than that of the control group (23). The results showed that in the mature solution, the mature solution, the mature solution and the semen were added to the EGCG, the percentage of cleavage was significantly higher than that of the control group (61.31%, 59.14%, 58.57% vs.45.71%, P0.05) in the mature solution, and the mature solution and the semen respectively (23). (23 .61%%, 22.77%, 23.56% vs.16.74%, P0.05). The above results show that EGCG can improve the fertilization ability of buffalo sperm and promote the development of early embryo, but there is no synergy between the addition of EGCG in the mature liquid and the semen. In order to be efficient and convenient, the best solution for this experiment is to add 10 u mol/L EGCG. to the mature solution or the semen, EGCG, EGCG. As a highly effective antioxidant, it can promote the maturation of buffalo oocyte in vitro, in vitro fertilization, and increase the development rate of the embryo in vitro,.EGCG has a positive effect on improving the production of buffalo embryos in vitro, and can improve the utilization of buffalo eggs. In the laboratory conditions, the effect of adding 10 mu mol/L EGCG to the mature solution or the semen is the most effective. OK.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S823.83
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