六味地黄苷糖对高糖诱导的胎盘滋养层细胞增殖侵袭及ERK/MMP-9信号通路的作用机制研究  

Study of Mechanism of LW-AFC on Proliferation and Invasion of Placental Trophoblast Cells Induced by High Glucose as well as on ERK/MMP-9 Signaling Pathway

作　　者：郭洁 王艺霏 张美子 王华莉 GUO Jie;WANG Yifei;ZHANG Meizi;WANG Huali

机构地区：[1]平煤神马医疗集团总医院妇幼体检科,河南平顶山467000 [2]上海复旦大学附属浦东医院康复科,上海201399

出　　处：《新中医》2021年第17期6-11,共6页New Chinese Medicine

基　　金：河南省医学科技项目(LHGJ20191525)。

摘　　要：目的:探究六味地黄苷糖(LW-AFC)通过调控ERK/MMP-9信号通路对高糖诱导的胎盘滋养层细胞增殖侵袭作用机制研究。方法:将在ATCC细胞库购买胎盘滋养层细胞分为空白对照组(空白组):胎盘滋养层细胞正常培养,不做任何处理;高糖诱导组(诱导组):采用高糖诱导的方式制作高糖胎盘滋养层细胞模型;六味地黄苷糖组(苷糖组):在高糖诱导胎盘滋养层细胞模型完成后给予LW-AFC培养。通过MTT法检测胎盘滋养层细胞增殖率,Western blot法检测胎盘滋养层细胞中ERK/MMP-9的蛋白含量,Transwell小室实验检测胎盘滋养层细胞侵袭情况,qRT-PCR法检测胎盘滋养层细胞中ERK/MMP-9mRNA的表达含量,克隆法检测胎盘滋养层细胞迁移情况的差异性,探究LW-AFC对高糖诱导的胎盘滋养层细胞增殖侵袭及ERK/MMP-9通路表达的影响。结果:随着时间的推移,诱导组胎盘滋养层细胞的增殖率降低,与诱导组比较,苷糖组细胞增殖率显著升高(P<0.05)。苷糖组与诱导组比较,苷糖组滋养层细胞侵袭能力显著高于诱导组细胞(P<0.05)。诱导组、苷糖组与空白组比较,空白组细胞克隆数量显著较多(P<0.05);苷糖组与诱导组比较,诱导组胎盘滋养层细胞克隆数量显著低于苷糖组(P<0.05)。诱导组胎盘滋养层细胞中ERK/MMP-9蛋白表达、ERK/MMP-9 mRNA表达量最高,空白组胎盘滋养层细胞蛋白表达、mRNA表达量低于苷糖组、诱导组(P<0.05),苷糖组与诱导组比较,苷糖组胎盘滋养层细胞ERK/MMP-9蛋白表达、ERK/MMP-9 m RNA表达量显著低于诱导组(P<0.05)。结论:LW-AFC能够提升高糖诱导的胎盘滋养层细胞的增殖速率,增加细胞迁移、侵袭的数量,LW-AFC的作用机制可能与下调ERK/MMP-9通路表达含量有关,这一试验结果可逐步应用于临床妊娠糖尿病患者的治疗中。Objective:To discuss the study of the mechanism of Liuwei Dihuang-active fraction combination(LW-AFC)on proliferation and invasion of placental trophoblast cells induced by high glucose by regulating ERK/MMP-9 signaling pathway. Methods:Placental trophoblast cells bought from ATCC cell bank were divided into the blank control group(the blank group),the high-glucose induction group(the induction group),and the Liuwei Dihuang-active fraction combination group(the glycoside group). In the blank group,placental trophoblast cells were normally cultured,without any treatment. In the induction group,placental trophoblast cell models with high glucose were established through high-glucose induction. After modeling,the placental trophoblast cell models with high glucose were cultured with LW-AFC in the glycoside group. The proliferation rate of placental trophoblast cells was detected by MTT method. The content of protein of ERK/MMP-9 in placental trophoblast cells was detected by Western blot method. The invasion of placental trophoblast cells was detected by Transwell chamber test. The expression level of ERK/MMP-9 mRNA in placental trophoblast cells was detected by qRT-PCR method. The differences in migration of placental trophoblast cells were detected by cloning method,so as to discuss the effect of LW-AFC on proliferation and invasion of placental trophoblast cells induced by high glucose as well as on expression of ERK/MMP-9 pathway. Results: With the passage of time, the proliferation rate of placental trophoblast cells in the induction group was decreased,and the rate in the glycoside group was significantly increased when compared with that in the induction group(P<0.05). The invasion ability of trophoblast cells in the glycoside group was significantly stronger than that in the induction group(P<0.05). The number of clones of placental trophoblast cells in the blank group was significantly larger than those in the induction group and the glycoside group(P<0.05),and the number in the induction group was signific

关 键 词：妊娠糖尿病 六味地黄苷糖 ERK MMP-9 胎盘滋养层细胞 

分 类 号：R285.5[医药卫生—中药学]