香烟烟雾提取物对小鼠卵母细胞和卵巢抗氧化酶表达的影响
发布时间：2019-01-02 07:08
【摘要】：【研究背景】全球范围内女性吸烟趋向年轻化，吸烟会导致女性妊娠率下降、自然流产率升高、活产率下降等。但香烟烟雾暴露对卵母细胞的影响及其机制仍然未明了。本研究通过观察香烟烟雾提取物（CSE）暴露对小鼠卵母细胞和卵巢抗氧化酶表达的影响及是否激活NRF2-ARE(核因子E2相关因子2-抗氧化反应序列元件)通路，尝试探索烟草暴露影响卵母细胞质量的机制，能为香烟烟雾暴露妇女的不孕症治疗提供基础依据。 【研究方法】 本研究采用C57BL/6小鼠测试CSE对获卵数、卵母细胞成熟度、形态和功能以及卵巢内抗氧化酶表达的影响。CSE组小鼠每日饮用CSE溶液（2mg/ml）；对照组饮用二甲基亚砜（DMSO）；阳性对照组每日腹腔注射15mg/kg的甲萘醌油溶液。分别处理4周。超排卵后取卵，观察获卵数、卵母细胞未成熟率、卵母细胞形态各参数和第一极体（PB）形态，取卵巢行qRT-PCR检测NRF2-ARE调控的抗氧化酶mRNA表达水平。 【结果】 （1）与对照组相比，，CSE组的去透明带卵母细胞直径（ZP-free OD）显著减小（对照组76.56±0.12，CSE组76.10±0.13，P 0.05），大小适中、圆形且表面光滑的第一极体（ARS-PB）率显著降低（对照组15.2%，CSE组6.6%，P 0.05），直径小、条状且表面粗糙的第一极体（SSR-PB）率显著增高（对照组7.6%，CSE组24.3%，P0.01）；（2）与阳性对照组相比，CSE组卵母细胞未成熟率显著降低（阳性对照组9.8%，CSE组3.3%，P 0.05），卵周间隙大小（PVS）显著减小（阳性对照组10.05μm，CSE组9.28μm，P 0.05）；SSR-PB率显著升高（CSE组24.3%，阳性对照组13.3%，P 0.05）。（3）与对照组相比，CSE组所检测NRF2-ARE通路中转录因子NRF2及其调控的8个抗氧化酶mRNA水平均无显著差异（P0.05）；（4）与CSE组对比，阳性对照组GSTM1（谷胱甘肽-硫-转移酶mu亚基1基因）表达水平显著降低（阳性对照组0.12±0.02，CSE组0.34±0.06，P 0.05）。 【结论】 结果提示：（1）CSE对卵母细胞有毒害作用；（2）甲萘醌生殖毒性表型与CSE不一致；（3）CSE、甲萘醌暴露，NRF2-ARE通路均无激活；（4）GSTM1在甲萘醌氧化应激模型中可能有其他通路抑制其表达。本研究为香烟烟雾暴露妇女的不孕症治疗提供了一些基础依据。
[Abstract]:Background: female smoking tends to be younger in the world. Smoking will lead to the decrease of pregnancy rate, natural abortion rate and live birth rate. However, the effect of cigarette smoke exposure on oocytes and its mechanism are still unclear. The aim of this study was to investigate the effects of cigarette smoke extract (CSE) on the expression of antioxidant enzymes in mouse oocytes and ovaries and whether to activate the NRF2-ARE (nuclear factor E2 related factor 2-antioxidant response sequence element) pathway. This paper attempts to explore the mechanism of tobacco exposure affecting oocyte quality, which can provide the basis for the treatment of infertility in women exposed to cigarette smoke. [methods] in this study, C57BL/6 mice were used to test the effects of CSE on the number of oocytes, oocyte maturity, morphology and function, and the expression of antioxidant enzymes in ovary. The CSE group mice drank CSE solution (2mg/ml) daily. The control group drank dimethyl sulfoxide (DMSO); positive control group daily intraperitoneal injection of 15mg/kg naphthoquinone oil solution. They were treated for 4 weeks respectively. After superovulation, the number of oocytes, the immature rate of oocytes, the parameters of oocyte morphology and the (PB) morphology of the first polar body were observed. The expression of antioxidant enzyme mRNA regulated by NRF2-ARE was detected by qRT-PCR in the ovary. [results] (1) compared with the control group, the diameters of the pellucida oocytes (ZP-free OD) in the CSE group were significantly decreased (76.56 卤0.12) (76.10 卤0.13), and the size was moderate. The rate of the first polar body (ARS-PB) with circular and smooth surface was significantly lower than that in the control group (control group 15.2cm), and the diameter of the first polar body was smaller than that of the control group (P 0.05). The rate of the first polar body (SSR-PB) with coarse surface was significantly higher than that of the control group (control group 7.6C: 24.3P0.01). (2) compared with the positive control group, the immature rate of oocytes in the CSE group was significantly lower than that in the positive control group (9.8% vs 3.3 渭 m, P 0.05), and the (PVS) in the peri-egg space was significantly decreased (10.05 渭 m in the positive control group). CSE group (9.28 渭 m); The SSR-PB rate was significantly higher in the CSE group than in the control group (24. 3% vs 13. 3%, P 0. 05). (3). There was no significant difference in the levels of transcription factor NRF2 and 8 antioxidant enzymes mRNA in NRF2-ARE pathway detected in CSE group (P0.05). (4) compared with CSE group, the expression of GSTM1 (glutathione-S-transferase mu subunit 1 gene) in positive control group was significantly decreased (0.12 卤0.02 卤0.34 卤0.06). [conclusion] the results suggest that: (1) CSE has toxic effect on oocytes, (2) the reproductive toxicity of menthoquinone is inconsistent with that of CSE, (3) CSE, exposure to menaphthoquinone does not activate the NRF2-ARE pathway; (4) the expression of GSTM1 may be inhibited by other pathways in the oxidative stress model of menthoquinone. This study provides some basis for the treatment of infertility in women exposed to cigarette smoke.
【学位授予单位】：广州医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.6
本文编号：2398162
[Abstract]:Background: female smoking tends to be younger in the world. Smoking will lead to the decrease of pregnancy rate, natural abortion rate and live birth rate. However, the effect of cigarette smoke exposure on oocytes and its mechanism are still unclear. The aim of this study was to investigate the effects of cigarette smoke extract (CSE) on the expression of antioxidant enzymes in mouse oocytes and ovaries and whether to activate the NRF2-ARE (nuclear factor E2 related factor 2-antioxidant response sequence element) pathway. This paper attempts to explore the mechanism of tobacco exposure affecting oocyte quality, which can provide the basis for the treatment of infertility in women exposed to cigarette smoke. [methods] in this study, C57BL/6 mice were used to test the effects of CSE on the number of oocytes, oocyte maturity, morphology and function, and the expression of antioxidant enzymes in ovary. The CSE group mice drank CSE solution (2mg/ml) daily. The control group drank dimethyl sulfoxide (DMSO); positive control group daily intraperitoneal injection of 15mg/kg naphthoquinone oil solution. They were treated for 4 weeks respectively. After superovulation, the number of oocytes, the immature rate of oocytes, the parameters of oocyte morphology and the (PB) morphology of the first polar body were observed. The expression of antioxidant enzyme mRNA regulated by NRF2-ARE was detected by qRT-PCR in the ovary. [results] (1) compared with the control group, the diameters of the pellucida oocytes (ZP-free OD) in the CSE group were significantly decreased (76.56 卤0.12) (76.10 卤0.13), and the size was moderate. The rate of the first polar body (ARS-PB) with circular and smooth surface was significantly lower than that in the control group (control group 15.2cm), and the diameter of the first polar body was smaller than that of the control group (P 0.05). The rate of the first polar body (SSR-PB) with coarse surface was significantly higher than that of the control group (control group 7.6C: 24.3P0.01). (2) compared with the positive control group, the immature rate of oocytes in the CSE group was significantly lower than that in the positive control group (9.8% vs 3.3 渭 m, P 0.05), and the (PVS) in the peri-egg space was significantly decreased (10.05 渭 m in the positive control group). CSE group (9.28 渭 m); The SSR-PB rate was significantly higher in the CSE group than in the control group (24. 3% vs 13. 3%, P 0. 05). (3). There was no significant difference in the levels of transcription factor NRF2 and 8 antioxidant enzymes mRNA in NRF2-ARE pathway detected in CSE group (P0.05). (4) compared with CSE group, the expression of GSTM1 (glutathione-S-transferase mu subunit 1 gene) in positive control group was significantly decreased (0.12 卤0.02 卤0.34 卤0.06). [conclusion] the results suggest that: (1) CSE has toxic effect on oocytes, (2) the reproductive toxicity of menthoquinone is inconsistent with that of CSE, (3) CSE, exposure to menaphthoquinone does not activate the NRF2-ARE pathway; (4) the expression of GSTM1 may be inhibited by other pathways in the oxidative stress model of menthoquinone. This study provides some basis for the treatment of infertility in women exposed to cigarette smoke.
【学位授予单位】：广州医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.6
【参考文献】
相关期刊论文 前1条
1 吴际,张丽珠;不同年龄妇女卵母细胞质量差异的分析[J];中华妇产科杂志;1998年07期
本文编号：2398162
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