摘要
目的研究青藤碱对复发性自然流产模型小鼠妊娠结局的影响及相关机制。方法雌性CBA/J小鼠60只,雄性DBA/2小鼠20只、Balb/c小鼠10只,按雌雄比例2∶1合笼,检出阴栓者计为妊娠第1天。分为正常组：雌性CBA/J小鼠×雄性Balb/c小鼠,正常饮食,9只孕鼠;模型组：雌性CBA/J小鼠×雄性DBA/2小鼠,正常饮食,10只孕鼠;治疗组：雌性CBA/J小鼠×雄性DBA/2小鼠,自检出阴栓日开始改用含有0.5%青藤碱的饲料,10只孕鼠。计数存活胚胎数和丢失胚胎数,并计算胚胎丢失率。使用ELISA法检测血清中细胞因子IFN-γ和IL-4的水平;real-time PCR检测IL-4、INF-γ、GATA-3、T-bet基因的表达水平;Western blot测IL-4、INF-γ、GATA-3、T-bet蛋白的表达水平。结果胚胎丢失率结果表明：与模型组比较,正常组和治疗组的胚胎丢失率显著降低（P〈0.01）。ELISA结果显示：与正常组相比较,模型组和治疗组中血清IFN-γ和IFN-γ/IL-4的水平均显著升高（P〈0.01）,IL-4水平均显著降低（P〈0.01）;与模型组相比较,治疗组中血清中IFN-γ水平和IFN-γ/IL-4比值均显著降低（P〈0.01）,而IL-4水平变化不大。Real-time PCR和Western blot的结果均显示：与正常组相比较,模型组和治疗组中IL-4、GATA-3基因的表达水平均显著降低（P〈0.01）;IFN-γ、T-bet基因的表达水平以及IFN-γ/IL-4比值、T-bet/GATA-3比值均显著升高（P〈0.01）。与模型组相比,治疗组中IFN-γ、T-bet基因的表达水平以及IFN-γ/IL-4比值、T-bet/GATA-3比值显著降低（P〈0.01）。结论青藤碱可以减少复发性自然流产的发生,其中RSA与Th1/Th2平衡漂移至Th1型有关;青藤碱治疗复发性自然流产的可能机制为降低体内Th1型细胞因子IFN-γ及转录因子T-bet的水平,从而调节Th1/Th2平衡。
Objective To investigate the influences and the mechanism of sinomenine on pregnancy outcome of mice with recurrent spontaneous abortion. Methods According to the proportion of female and male 2∶1 cage,60 female CBA / J mice,20 male DBA /2 mice,10 Balb / c mice,it was detection of pessary which counted as first day of pregnancy. It was divided into three group,the normal group： female CBA / J mice × male Balb / c mice,normal diet,9 pregnant mice; Model group： female CBA / J mice × male DBA /2 mice,normal diet,10 pregnant mice; treatment group： female CBA / J mice × male DBA /2 mice,when self checking out the vaginal plug day,the mice were fed with containing 0. 5% of sinomenine,10 pregnant mice. Counting the number of embryos and embryos survive loss,and calculated the rate of embryo loss. ELISA method was used for the detection of serum cytokines,IFN-γ and IL-4 level; the expression level of IL-4,INF-γ,GATA-3,T-bet gene were detected by real-time PCR; the level of IL-4,INF-γ,GATA-3,T-bet protein were measured by Western Blot method. Results Rate of embryo loss results showed that： compared with the model group,the rate of embryo loss of the normal group and the treatment group was decreased significantly（ P〈0. 01）,the difference was statistically significant. ELISA results showed that： compared with the normal group,model group and treatment group in serum IFN-γ and IFN-γ / IL-4 levels were significantly increased（ P〈0. 01）,the levels of IL-4 were significantly decreased（ P〈0. 01）,the difference was statistically significant; compared with the model group,the treatment group in serum IFN-rlevel and the ratio of IFN-γ / IL-4 were significantly decreased（ P〈0. 01）,but the level of IL-4changed little,no significant difference. The results of real-time PCR and Western blot indicated that compared with the normal group,the expression level of IL-4 and GATA-3 genes in model group and treatment group were significantly decreased（ P〈0. 01）; the expression level of IFN-�
出处
《中国生化药物杂志》
CAS
2015年第1期63-67,共5页
Chinese Journal of Biochemical Pharmaceutics
基金
山东省自然科学基金课题项目（ZR2009CL027）
济南市科技局科技明星计划课题项目（20100118）
山东省医学科学院医学基金课题项目（201023）
