五子衍宗丸预防神经管畸形的机制研究

Mechanism of Wuzi Yanzong Pill in Preventing Neural Tube Defects

目的:观察五子衍宗丸(WYP)与叶酸(FA)对神经管畸形(NTDs)的预防作用并进行比较,研究五子衍宗丸防控神经管畸形的氧化应激机制。方法:C57BL/6小鼠36只,按雌雄=2∶1合笼,检测到阴栓者为孕鼠,并将其随机分为正常组、模型组、叶酸组和五子衍宗丸组每组各6只,同时给予药物干预,孕7.5 d采用全反式维甲酸(atRA)建立NTDs模型(正常组除外),孕11.5 d处死孕鼠取出子宫分离胚胎,体视显微镜下鉴定神经管畸形。氧化应激试剂盒检测胚胎组织还原型谷胱甘肽(GSH)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活力,蛋白质免疫印迹法检测胚胎组织过氧化氢酶(CAT)蛋白表达。结果:镜下观察小鼠胚胎,正常组神经管闭合良好,模型组呈现神经管畸形。与正常组比较模型组GSH、CAT、SOD和GSH-Px表达量显著降低;与模型组比较叶酸组和五子衍宗丸组GSH、CAT、SOD和GSH-Px的表达呈不同程度的增高;与叶酸组比较五子衍宗丸组GSH、CAT、SOD和GSH-Px的表达均增高。结论:五子衍宗丸和叶酸可以明显改善胎鼠神经管的闭合情况,降低NTDs发病率,且五子衍宗丸的预防效果优于叶酸,其作用机制可能为上调GSH、CAT、SOD和GSH-Px的表达水平,抑制氧化应激反应。

Objective:To observe and compare the effects of Wuzi Yanzong pill(WYP)and folic acid(FA)on Neural tube defects(NTDs)as well as the oxidative stress mechanism of WYP in preventing and controlling NTDs.Methods:Thirty six C57BL/6 mice were caged according to the ratio of female to male=2∶1,and vaginal plugs were detected as pregnant mice.Pregnant mice were randomly divided into normal group,model group,FA group and WYP group with 6 mice in each group,and drug intervention was given at the same time.NTDs model was established by all-trans-retinoic acid(atRA)on day 7.5 of gestation(except normal group).On day 11.5 of pregnancy,the embryo was separated in uterus,and NTDs was identified under stereomicroscope.The content of Glutathione(GSH)and the activity of Superoxide Dismutase(SOD)and Glutathione peroxidase(GSH-Px)in embryonic tissues were detected by oxidative stress kit.The protein expression of Catalase(CAT)in embryonic tissues was detected by Western Blot.Results:Under the microscope,the neural tube of the normal group was well closed,and the model group was deformed.Compared with the normal group,the expression of GSH,CAT,SOD and GSH-Px in the model group were significantly reduced;compared with the model group,the expression of GSH,CAT,SOD and GSH-Px in FA group and WYP group were increased in different degrees;compared with FAgroup,the content of GSH,CAT,SOD and GSH-Px in WYP group were increased.Conclusion:WYP and FA can significantly improve the closure of fetal mice neural tube and reduce the incidence of NTDs,and the preventive effects of WYP is better than that of FA.The mechanism may be to up-regulate the expression of GSH,CAT,SOD and GSH-Px and inhibit oxidative stress.