1.河北中医药大学中西医结合学院 石家庄 050091
2.河北中医药大学基础医学院
3.河北省中西医结合生殖疾病协同创新中心
4.河北省中西医结合肝肾病证研究重点实验室
唐思玲,女,在读硕士生
# 段彦苍,女,博士,教授,博士生导师,主要研究方向:中西医结合生殖内分泌,E-mail:duanyancang@163.com
纸质出版日期:2023-08-30,
网络出版日期:2023-05-12,
收稿日期:2022-09-15,
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唐思玲, 梁潇, 李荣霞, 等. 寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响[J]. 北京中医药大学学报, 2023,46(8):1139-1149.
TANG Siling, LIANG Xiao, LI Rongxia, et al. Effects of
唐思玲, 梁潇, 李荣霞, 等. 寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响[J]. 北京中医药大学学报, 2023,46(8):1139-1149. DOI: 10.3969/j.issn.1006-2157.2023.08.012.
TANG Siling, LIANG Xiao, LI Rongxia, et al. Effects of
目的
2
探讨寿胎丸对复发性流产(RSA)小鼠胚胎滋养细胞上皮间质转化(EMT)及胚胎丢失率的影响。
方法
2
将90只6~8周龄CBA/J雌性小鼠按随机数字表法分为正常组、模型组、地屈孕酮组和寿胎丸低、中、高剂量组,每组15只;正常组与BALB/c雄鼠合笼建立正常妊娠小鼠,其他组雌鼠与DBA/2雄鼠合笼建立RSA小鼠模型。合笼前14 d开始每日给药,寿胎丸低、中、高剂量组分别以寿胎丸7.58、15.17、30.33 g/kg灌胃,地屈孕酮组以地屈孕酮0.003 g/kg灌胃,正常组和模型组以等体积蒸馏水灌胃。连续给药至妊娠第6天(PD 6),各组随意选取9只孕鼠麻醉处死后剥离胚胎,HE染色观察PD 6小鼠母胎界面滋养细胞病理形态学改变;免疫荧光染色、实时荧光定量PCR检测各组胚胎滋养细胞EMT标志物上皮钙黏素(E-cadherin)、波形蛋白(Vimentin)及β-连环蛋白(β-catenin)的蛋白及mRNA表达。各组其余孕鼠继续给药至妊娠第14天(PD 14),麻醉处死后观察胚胎丢失情况。
结果
2
与正常组比较,模型组孕鼠胚胎滋养细胞中上皮细胞标记物E-cadherin蛋白及mRNA表达升高(
P
<
0.05),间质细胞标志物Vimentin蛋白及mRNA表达下降(
P
<
0.05)、β-catenin蛋白及mRNA表达下降(
P
<
0.05),胚胎丢失率升高(
P
<
0.05);与模型组比较,各剂量寿胎丸组及地屈孕酮组胚胎滋养细胞中E-cadherin蛋白及mRNA表达下调(
P
<
0.05),Vimentin蛋白及mRNA表达升高、β-catenin蛋白及mRNA表达升高(
P
<
0.05),其中寿胎丸中、高剂量组及地屈孕酮组更为明显,寿胎丸低剂量组胚胎丢失率有所降低(
P
>
0.05),寿胎丸中、高剂量组及地屈孕酮组胚胎丢失率下降(
P
<
0.05)。
结论
2
寿胎丸能降低RSA小鼠胚胎丢失率,其机制可能是通过激活滋养细胞内β-catenin信号分子,促进EMT改善胚胎滋养细胞迁移侵袭能力,以利于胚胎在宫腔内着床而发挥保胎作用。
Objective
2
We aimed to explore the effect of
Shoutai
Pills on the epithelial-mesenchymal transition (EMT) of trophoblasts and the embryo loss rate in embryos of mice with recurrent spontaneous abortion (RSA).
Methods
2
Ninety CBA/J female mice aged 6-8 weeks old were randomly divided into the normal group
the model group
the dydrogesterone group
the
Shoutai
Pills low-dose group
the
Shoutai
Pills medium-dose group and the
Shoutai
Pills high-dose group with 15 mice in each group. The normal pregnancy mouse was established with the mice in normal group × BALB/c male mice
and the RSA mice model was established with the mice in other groups × DBA/2 male mice. Female mice were administered doses 14 days before mating. The
Shoutai
Pills low-dose
medium-dose and high-dose groups were given 7.58
15.17
30.33 g/kg intragastric administration respectively
and dydrogesterone group was given dydrogesterone 0.003 g/kg intragastric administration
the normal group and the model group were given the same volume of distilled water intragastric administration. After continuous dosing to the 6th day of pregnancy
nine pregnant mice were selected from each group randomly and sacrificed under anesthesia
and then the embryos were harvested. Pathological changes of trophoblast cells at the maternal-fetal interface were observed by hematoxylin-eosin staining. Immunofluorescence staining was used to determine the protein expression levels
and real-time fluorescence quantitative PCR was used to determine the mRNA expression levels of EMT markers (E-cadherin
Vimentin) and β-catenin of embryonic trophoblasts in each group. The remaining pregnant mice in each group were administered doses until the 14th day of pregnancy
and after mice were sacrificed under anesthesia
embryo loss in each group was observed.
Results
2
Compared with the normal group
the protein and mRNA expression levels of the epithelial cell marker E-cadherin in the embryonic trophoblasts of the model group were increased (
P
<
0.05)
the expression levels of the mesenchymal cell marker Vimentin in the embryonic trophoblasts of the model group were decreased(
P
<
0.05)
the expression levels of β-catenin in the embryonic trophoblasts were decreased (
P
<
0.05)
and the embryo loss rate of the model group was increased (
P
<
0.05). Compared with the model group
the expression levels of E-cadherin in the embryonic trophoblasts were decreased (
P
<
0.05)
the expression levels of Vimentin in the embryonic trophoblasts were increased
and the expression levels of β-catenin in the embryonic trophoblasts were increased of each dose
Shoutai
Pills group and dydrogesterone group(
P
<
0.05)
all of which were most significant in the
Shoutai
Pills medium-dose and high-dose groups and the dydrogesterone group
the embryo loss rate in the
Shoutai
Pills low-dose group was decreased (
P
>
0.05)
and the embryo loss rate in the
Shoutai
Pills medium-dose and high-dose groups and the dydrogesterone group was decreased significantly (
P
<
0.05) .
Conclusion
2
Shoutai
Pills can reduce the rate of embryo loss in RSA mice
possibly by promoting EMT by activating β-catenin signaling molecules in trophoblasts and improving the migration and invasion ability of embryonic trophoblasts. These effects are conducive to the implantation of the embryo in the uterine cavity and play a role in protecting the fetus.
复发性流产寿胎丸滋养细胞上皮间质转化小鼠
recurrent spontaneous abortionShoutai Pillstrophoblastepithelial-mesenchymal transitionmice
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