高糖培养环境对小鼠胚胎发育潜能的影响  

Effects of exposure to high glucose concentration on embryonic development potential of mouse

作　　者：文亮 陈书强 郭翔宇 董杰 雷晖 马媛 王晓红 Wen Liang;Chen Shuqiang;Guo Xiangyu;Dong Jie;Lei Hid;Ma Yuan;Wang Xiaohong(Reproductive Medical Centre, Tangdu Hospital, the Fourth Military Medical University, Xi'an 710038, China)

机构地区：[1]空军军医大学唐都医院生殖医学中心,西安710038

出　　处：《中华生殖与避孕杂志》2019年第5期370-375,共6页Chinese Journal of Reproduction and Contraception

基　　金：国家自然科学基金-面上项目(81671463);陕西省重点研发计划重点项目(2017ZDCXLSF-02-03).

摘　　要：目的评估植入前高糖培养环境对小鼠胚胎发育潜能的影响。方法选择SPF级雌性ICR小鼠,促排卵后与正常雄鼠交配获取原核胚,分别在含0.2 mmol/L葡萄糖的KSOM培养液(对照组)、含10 mmol/L葡萄糖的KSOM培养液(中剂量组)、含20 mmol/L葡萄糖的KSOM培养液(高剂量组)培养至囊胚,每组15只雌性小鼠,记录3组桑葚胚率及囊胚率;3组囊胚移植入第3.5日(D3.5)代孕母鼠子宫,于D18.5处死母鼠,计算胚胎植入率、成活率、胚胎停育率;提取3组小鼠囊胚mRNA,采用实时荧光定量聚合酶链式反应(RT-qPCR)检测细胞命运决定因子以及植入和早期血管发生相关基因的mRNA表达量。结果①与对照组相比,高剂量组囊胚形成率显著降低(P=0.016)。②3组胚胎植入率差异无统计学意义(P>0.05),但与对照组相比,中剂量组和高剂量组植入后胚胎成活率显著降低(P=0.004),胚胎停育率显著增高(P=0.004)。③与对照组相比,中剂量组和高剂量组早期细胞命运决定因子Cdx2、Tead4、Fgfr2相对表达水平均显著下调(P<0.001,P=0.005,P=0.003)。④与对照组相比,中剂量组和高剂量组囊胚期胚胎植入及早期血管发生相关基因Adm、Plau、Timp1相对表达水平均显著下调(P<0.001,P=0.001,P=0.041),Mmp2、Timp3、Troap相对表达水平均显著上调(P=0.012,P=0.013,P=0.002)。结论植入前胚胎处于高糖培养环境,降低了胚胎的发育潜能,并导致了不良妊娠结局发生。Objective To evaluate the effect of exposure to high glucose concentration on the developmental potential of preimplantation embryos. Methods Female ICR mice mated with males after ovulation induction to acquire zygotes, which were incubated to blastocysts in KSOM medium (0.2 mmol/L glucose, control group), KSOM medium supplemented with 10 mmol/L glucose (middle dose group) and KSOM medium supplemented with 20 mmol/L glucose (high dose group). The blastocyst rate of each group was recorded. The blastocysts were transplanted into day 3.5 (D3.5) pseudopregnant mice. The surrogate female mice were sacrificed in D18.5. Then, the embryo implantation rate, the live fetal rate after implantation and the embryo abortion rate were calculated. The blastocysts mRNA were extracted and the mRNA expression levels of relevant genes (Cdx2, Tead4, Fgfr2, Adm, Plau, Timp1) were detected by real-time quantitative polymerase chain reaction. Results 1) Compared with control group, the blastocyst rate of the high dose group was significantly decreased (P=0.016). 2) There was no statistical difference in the implantation rate of the three groups (P>0.05), but the live fetal rate after implantation of the middle dose group and high dose group was significantly decreased compared with control group (P=0.004). 3) Compared with control group, the mRNA relative expression levels of Cdx2, Tead4 and Fgfr2 were significantly down-regulated (P<0.001, P=0.005, P=0.003) in the blastocysts of middle and high dose groups. 4) Compared with control group, the mRNA relative expression levels of Adm, Plau and Timp1 were significantly down-regulated (P<0.001, P=0.001, P=0.041), and Mmp2, Timp3 and Troap were significantly up-regulated in the blastocysts of middle and high dose groups (P=0.012, P=0.013, P=0.002). Conclusion Before implantation, exposure to high glucose concentration decreases the developmental potential of the embryos, and leads to adverse pregnant outcomes.

关 键 词：高糖 囊胚 植入率 胚胎停育率 葡萄糖代谢 

分 类 号：R714.8[医药卫生—妇产科学]