摘要
为研究妊娠合并糖尿病对孕妇及胎儿产生危害的机制,构建妊娠合并糖尿病的昆明小鼠动物模型,检测不同浓度葡萄糖对体外培养胚泡细胞生长的影响.观察不同浓度胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)对体外高糖环境胚泡细胞发育的影响,利用细胞核DNA双染实验观察不同浓度IGF-1作用下胚泡细胞的凋亡.采用实时定量PCR(RT-PCR)分析不同凋亡相关基因在体外培养胚泡中的表达情况.结果显示,随着葡萄糖浓度的增加,胚泡细胞总数减少,高浓度葡萄糖(≥30mmol/L)则能显著性抑制胚泡细胞的生长(P<0.01).RT-PCR检测发现妊娠合并糖尿病小鼠的胚泡igf-1表达下调,且与葡萄糖的浓度成正相关.凋亡相关基因bcl-2和bcl-xl的表达随着体外IGF-1培养浓度的增加而表达上调,而p53基因和凋亡相关基因Bax的表达则下调.细胞凋亡实验显示,随着IGF-1浓度的增加,体外培养胚泡细胞的凋亡逐渐降低,当IGF-1浓度达到100μg/L时,几乎未发现细胞凋亡.因此,高血糖能抑制胚泡细胞的生长发育,导致igf-1的表达下调,而IGF-1能抑制胚泡细胞的凋亡,有利于胚泡细胞的生长发育.
In order to investigate the mechanisms of pregnancy associated with diabetes harm to gravida and fetal, The Kunming mice models for pregnancy associated with diabetes were simulated, The effects of glucose in different concentrations was detected on the growth of embryo cells in vitro culture. The effects of IGF-1 in different concentrations was determined on the development ofblastocyst in hyperglycemic conditions （30.0 mol/L glucose） in vitro, as well as the apoptosis of embyro cells by using nuclear DNA double-dyed assays. Moreover, the expression of igf-1 and other genes associated with apoptosis in vitro embryo was detcted by Real-time quantitative PCR. The results showed that the total cell number of blastocyst fell off along with the increasing of glucose concentration. High concentrations of glucose （≥ 30 mmol/L） could significantly inhibit the growth of embryo cells （P 〈 0.01）. The results of real-time quantitative PCR showed that the expression of igf-1 was down-regulated in mouse blastocyst with pregnancy associated with diabetes, and positively correlated with the concentration of glucose. The expressions of apoptosis-related genes bcl-2 and bcl-xl were up-regulated along with the increasing oflGF-1 concentration, while the p53 gene and the apoptosis-related gene Bax were down-regulated. The embryo cells apoptosis had been gradually reduced with the increasing of IGF-I concentration in vitro, and in the IGF-1 concentration of 100 μg/L, there was almost no apoptosis in embryo cells. These experiments demonstrated that hyperglycemic conditions in vitro can inhibit the growth and development of the embryo cells resulting in the down-regulated expression of igf-1, and IGF-1 could inhibit the apoptosis of blastocyst and be benefit for the growth of blastocyst.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2009年第6期776-780,共5页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金资助项目(30600224,30700438,30771135)
中国博士后科学基金(20060390886)
湖南省自然科学基金(06JJ30014)
湖南省科技计划(2008FJ3138)
湖南省教育厅科研资助项目(07B008)~~
关键词
妊娠合并糖尿病
鼠胚胎
体外培养
IGF-1
凋亡
pregnancy associated with diabetes, mouse embryos, in vitro culture （IVC）, IGF-1, apoptosis
