【摘要】：【研究背景】全球范圍內(nèi)女性吸煙趨向年輕化，吸煙會導致女性妊娠率下降、自然流產(chǎn)率升高、活產(chǎn)率下降等。但香煙煙霧暴露對卵母細胞的影響及其機制仍然未明了。本研究通過觀察香煙煙霧提取物（CSE）暴露對小鼠卵母細胞和卵巢抗氧化酶表達的影響及是否激活NRF2-ARE(核因子E2相關(guān)因子2-抗氧化反應序列元件)通路，嘗試探索煙草暴露影響卵母細胞質(zhì)量的機制，能為香煙煙霧暴露婦女的不孕癥治療提供基礎(chǔ)依據(jù)。 【研究方法】 本研究采用C57BL/6小鼠測試CSE對獲卵數(shù)、卵母細胞成熟度、形態(tài)和功能以及卵巢內(nèi)抗氧化酶表達的影響。CSE組小鼠每日飲用CSE溶液（2mg/ml）；對照組飲用二甲基亞砜（DMSO）；陽性對照組每日腹腔注射15mg/kg的甲萘醌油溶液。分別處理4周。超排卵后取卵，觀察獲卵數(shù)、卵母細胞未成熟率、卵母細胞形態(tài)各參數(shù)和第一極體（PB）形態(tài)，取卵巢行qRT-PCR檢測NRF2-ARE調(diào)控的抗氧化酶mRNA表達水平。 【結(jié)果】 （1）與對照組相比，，CSE組的去透明帶卵母細胞直徑（ZP-free OD）顯著減�。▽φ战M76.56±0.12，CSE組76.10±0.13，P 0.05），大小適中、圓形且表面光滑的第一極體（ARS-PB）率顯著降低（對照組15.2%，CSE組6.6%，P 0.05），直徑小、條狀且表面粗糙的第一極體（SSR-PB）率顯著增高（對照組7.6%，CSE組24.3%，P0.01）；（2）與陽性對照組相比，CSE組卵母細胞未成熟率顯著降低（陽性對照組9.8%，CSE組3.3%，P 0.05），卵周間隙大�。≒VS）顯著減小（陽性對照組10.05μm，CSE組9.28μm，P 0.05）；SSR-PB率顯著升高（CSE組24.3%，陽性對照組13.3%，P 0.05）。（3）與對照組相比，CSE組所檢測NRF2-ARE通路中轉(zhuǎn)錄因子NRF2及其調(diào)控的8個抗氧化酶mRNA水平均無顯著差異（P0.05）；（4）與CSE組對比，陽性對照組GSTM1（谷胱甘肽-硫-轉(zhuǎn)移酶mu亞基1基因）表達水平顯著降低（陽性對照組0.12±0.02，CSE組0.34±0.06，P 0.05）。 【結(jié)論】 結(jié)果提示：（1）CSE對卵母細胞有毒害作用；（2）甲萘醌生殖毒性表型與CSE不一致；（3）CSE、甲萘醌暴露，NRF2-ARE通路均無激活；（4）GSTM1在甲萘醌氧化應激模型中可能有其他通路抑制其表達。本研究為香煙煙霧暴露婦女的不孕癥治療提供了一些基礎(chǔ)依據(jù)。
[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 鹵0.12) (76.10 鹵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 鹵0.02 鹵0.34 鹵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.
【學位授予單位】：廣州醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R711.6

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