Study of Mechanism of LW- AFC on Proliferation and Invasion of Placental Tropho⁃ blast Cells Induced by High Glucose as well as on ERK/MMP-9 Signaling Pathway
Abstract: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 significantly smaller than that in the glycoside group(P<0.05). The expression levels of mRNA and protein of ERK/MMP- 9 of placental trophoblast cells in the induction group was the highest among the three groups,and the expression levels in the blank group were lower than those in the induction group and the glycoside group(P<0.05). The expression levels of mRNA and protein of ERK/MMP-9 of placental trophoblast cells in the glycoside group were significantly lower than those in the induction group(P< 0.05). Conclusion:LW- AFC can promote the proliferation rate of placental trophoblast cells induced by high glucose,and increase the number of migration and invasion of cells. Its mechanism is probably related to down- regulation of expression levels of ERK/MMP- 9 pathway. The experimental results can be gradually applied in the clinical treatment of patients with gestational diabetes mellitus.