寿胎丸对复发性流产模型小鼠胚胎有氧糖酵解关键酶及相关蛋白表达的影响  

Effects of Shoutai Pills(寿胎丸)on the Expression of Key Enzymes and Related Proteins of Aerobic Glycolysis in Embryos of Recurrent Spontaneous Abortion Model Mice

作　　者：梁潇 唐思玲 宋亚静 李丹丹 段彦苍 杜惠兰 LIANG Xiao;TANG Siling;SONG Yajing;LI Dandan;DUAN Yancang;DU Huilan(College of Integrated Traditional Chinese and Western Medicine,Hebei University of Chinese Medicine,Shijiazhuang,050091;Hebei Collaborative Innovation Center of Integrated Chinese and Western Medicine on Reproductive Disease)

机构地区：[1]河北中医学院中西医结合学院,050091 [2]河北省中西医结合生殖疾病协同创新中心

出　　处：《中医杂志》2023年第10期1044-1050,共7页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(U21A20403);河北省创新能力提升计划(20567624H);河北中医学院研究生创新资助项目(CXZZBS2022094)。

摘　　要：目的 探讨寿胎丸治疗复发性流产(RSA)的可能作用机制。方法 雌性CBA/J小鼠随机分为正常组、模型组、地屈孕酮组和寿胎丸组,每组14只。正常组雌性CBA/J小鼠与雄性BALB/c小鼠合笼交配;其余3组为雌性CBA/J小鼠与雄性DBA/2小鼠合笼交配建立RSA小鼠模型。寿胎丸组雌鼠从合笼前14天到妊娠第6天或第14天给予寿胎丸1.66 g/(kg·d)灌胃;地屈孕酮组妊娠第1天到妊娠第6天或第14天给予地屈孕酮片0.33 g/(kg·d)灌胃,正常组和模型组雌鼠给予0.4 ml蒸馏水灌胃。各组在妊娠第6天时随机选取8只雌鼠取胚胎组织,检测胚胎组织糖酵解关键酶己糖激酶2 (HK2)、M2-型丙酮酸激酶(PKM2)、乳酸脱氢酶A (LDHA) mRNA和蛋白表达,检测葡萄糖转运体1 (GLUT1)、单羧酸转运体4 (MCT4)和G蛋白偶联受体81 (GPR81)蛋白表达水平。剩余6只雌鼠在妊娠第14天注射台盼蓝染色剂,计算胚胎丢失率。结果 妊娠第14天,正常组雌鼠胚胎丢失率为6.12%,模型组小鼠胚胎丢失率为33.33%,差异有统计学意义(P<0.05);地屈孕酮组和寿胎丸组胚胎丢失率较模型组显著降低,分别为12.24%和14.29%(P<0.05)。妊娠第6天,与正常组比较,模型组小鼠胚胎HK2、PKM2和LDHA mRNA及蛋白表达水平降低(P<0.05),GLUT1、MCT4和GPR81蛋白表达水平降低(P<0.05);与模型组比较,地屈孕酮组除LDHA蛋白表达外其余各指标表达水平均显著升高(P<0.05),寿胎丸组除PKM2 mRNA表达外其余各指标亦均升高(P<0.05);与地屈孕酮组比较,寿胎丸组小鼠胚胎MCT4蛋白表达水平升高,GPR81蛋白表达水平降低(P<0.05)。结论 寿胎丸可能通过促进胚胎有氧糖酵解关键酶及相关蛋白的表达,从而提高有氧糖酵解水平,降低胚胎丢失率,进而发挥其保胎作用。Objective To explore the possible mechanism of Shoutai Pill(寿胎丸)in the treatment of recurrent spontaneous abortion(RSA).Methods Female CBA/J mice were randomly divided into normal group,model group,dydrogesterone group and Shoutai Pill group,with 14 mice in each group.Female CBA/J mice were co-mated with male BALB/c mice in the normal group,while those were co-mated with male DBA/2 mice to establish an RSA model group in the other three groups.Female mice in Shoutai Pill group were given 1.66 g/(kg·d)of Shoutai Pill by gavage from 14 days before mating until the 6th or 14th day of pregnancy.The dydrogesterone group was given 0.33 g/(kg·d)of dydrogesterone by gavage from the 1st day to the 6th or 14th day of pregnancy,while the normal group and the model group were given 0.4 ml of distilled water by gavage.On the 6th day of pregnancy,eight female rats were randomly selected from each group to take embryonic tissues,and the mRNA and protein expressions of key enzymes of aerobic glycolysis in embryos including hexokinase 2(HK2),pyruvate kinase M2(PKM2),and lactate dehydrogenase A(LDHA)were detected,as well as the protein expression of glucose transporter 1(GLUT1),monocarboxylic acid transporter 4(MCT4)and G protein-coupled receptors 81(GPR81).The remaining six female mice in each group were anesthetized on the 14th day of pregnancy,injected with trypan blue dye,and the embryo loss rate(ELR)was calculated.Results On the 14th day of pregnancy,the ELR of female mice in the normal group was 6.12%,and that in the model group was 33.33%,with statistically significant difference between the groups(P<0.05).The ELR of female mice in the dydrogesterone group and Shoutai Pill group was 12.24%and 14.29%,respectively,significantly lower than that in the model group(P<0.05).On the 6th day of pregnancy,compared to those in the normal group,the mRNA and protein expression levels of HK2,PKM2 and LDHA in the model group decreased(P<0.05),as well as the protein expression levels of GLUT1,MCT4 and GPR81(P<0.05).Compared to thos

关 键 词：复发性流产 寿胎丸 有氧糖酵解 胚胎丢失率 

分 类 号：R285.5[医药卫生—中药学]