分泌型晚期糖基化终产物受体对妊娠糖尿病胚胎保护的实验研究

摘要

目的 研究分泌型晚期糖基化终产物受体(sRAGE)在实验动物罹患妊娠糖尿病(GDM)时的表达差异及对子鼠发育的影响.方法 采用给SD孕鼠空腹一次性腹腔注射链脲佐菌素(STZ) 25mg/kg的方法建立宫内高血糖孕鼠模型.STZ注射后对小鼠随机分为实验组和对照组,每间隔24小时,实验组孕鼠行尾静脉注射重组sRAGE蛋白(5mg/kg,0.2 ml PBS)即为sRAGE组,对照组注射相同剂量的白蛋白溶液.分别于第3、13、19天检测孕鼠血糖、血清AGEs及脑、心脏组织中的RAGE蛋白水平.最后一日剖宫取胎,剥离胎盘,检测胎盘RAGE、NOX2、MCP-1、VCAM-1的mRNA表达水平,探讨血清中AGEs、RAGE表达水平与胚胎发育的关系.结果 实验组孕鼠血清中血糖和AGEs的浓度显著高于对照组(P＜0.05),相关性分析显示血糖与血清中AGEs水平呈显著正相关(r=0.693,P＜0.05);与正常对照组相比,实验组孕鼠的胎盘、脑及心脏组织中活性氧产生,炎症相关基因的mRNA和RAGE蛋白水平显著升高,而sRAGE处理可以部分降低这些活性氧产生、炎症相关基因的表达和RAGE的表达.对照组孕鼠胎盘、脑以及心脏组织中的RAGE蛋白表达极弱,实验组在3种组织中的RAGE表达则明显增强.在sRAGE注射干预后,RAGE蛋白表达与实验组相比显著降低,仍高于对照组.结论 sRAGE静脉注射可显著降低孕鼠子代组织中NF-κB的活性,通过调控相关细胞因子发挥对胚胎的保护作用.%Objective To study the differential expression of the receptor for advanced glycation end products in the experimental animals with gestational diabetes mellitus and the effect on the development of their offspring.Methods The method of abdominal cavity injection of SD (Streptozotocin,STZ) 25mg/Kg was used to establish the rat model of intrauterine hyperglycemia.The mice after STZ injection were randomly divided into experimental group of pregnant rats treated with tail vein injection of recombinant sRAGE protein (5 mg/kg,0.2 ml PBS) and control group injected with the same dose of albumin solution.Blood glucose,serum AGEs,and RAGE protein levels in brain and heart tissue were detected at 3,13 and 19 days,respectively.On the last day of cesarean section;stripping the placenta,the placental RAGE,NOX2,VCAM-1,mRNA MCP-1 expression levels,to explore the relationship between serum AGEs,RAGE expression levels and the development of the embryo were observed.Results The concentration of blood glucose and AGEs in serum of GDM rats were significantly higher than that of control group (P＜0.05).The correlation analysis showed significantly positive correlation between AGEs and serum glucose level (r=0.693,P＜0.05).The heart tissue oxidative stress,the expression level of mRNA and RAGE proteins of inflammation related genes of GDM group significantly increased compared with those of control group.sRAGE could reduce the expression of RAGE and part of the production of reactive oxygen species and the inflammation related genes.The placenta of pregnant rats,brain and heart tissue and the expression of RAGE protein expression in GDM group were very weak and RAGE significantly increased.With the combined injection of sRAGE intervention,the expression of RAGE protein was significantly decreased compared with GDM group,but still higher than the normal control group.Conclusion SRAGE intravenous injection can significantly reduce the pregnant offspring tissue NF-kappa B activity and exert a protective effect on embryos by regulating cytokines.