Total flavonoids from Semen Cuscutae target MMP9 and promote invasion of EVT cells via Notch/AKT/MAPK signaling pathways

Abstract

Miscarriage is a common condition during pregnancy and its mechanisms remain largely unknown. Extravillous trophoblast (EVT) cell invasion is required to maintain normal pregnancy and its malfunction has been proposed as a major cause for miscarriage. Homeostasis of matrix metalloproteinase 9 (MMP9) is a key to regulate EVT cell invasion. Total flavonoids from Semen Cuscutae (TFSC) have been applied clinically used for preventing or treating miscarriage in the past. Given its potential clinical benefit on preventing miscarriage, this study aims at examining the therapeutic effect of TFSC in the prevention of premature birth by upregulating MMP9 and promote EVT cell invasion. HTR-8 cells migration and invasion functions were analyzed using wound healing and transwell assays. The regulatory effect of TFSC on MMP9 expression and relevant signaling pathways were analyzed by Western Blot. The results show compared to control group, TFSC significantly promoted the migration of EVT cells in a dose and time-dependent manner. The migration and invasion of EVT cells were maximized at the highest dosage of 5 μg/ml of TFSC. The expression of MMP9 in EVT cells was significantly increased after TFSC treatment. Furthermore, cells treated with TFSC significantly upregulated protein expressions in Notch, AKT and p38/MAPK signaling pathways. We believe TFSC can promote the migration and invasion of EVT cells by increasing MMP9 expression, and prevent miscarriage by activating Notch, AKT, and MAPK signaling pathways.

MNSFβ Promotes the Proliferation and Migration of Human Extravillous Trophoblast Cells and the Villus Expression Level of MNSFβ Is Decreased in Recurrent Miscarriage Patients

“…In this study, the positive correlations among the expression levels of MNSFβ, P53, and RhoA, as well as the proliferation and migration abilities of HTR8/SVneo cells were observed, indicating that the P53-RhoA pathway might be involved in the human trophoblast cell proliferation and migration and MNSFβ might promote EVT cell proliferation and migration by activating the P53-RhoA pathway. MMP-9 plays critical roles in regulation of EVT cell invasion, and an increased MMP-9 expression was found to be accompanied with the enhanced EVT cell migration [26]. More interestingly, it was reported that simultaneously decreased expression levels of MMP9 and RhoA were detected in trophoblast cells with reduced invasion ability and MMP9 could regulate trophoblast invasion ability through Rho-family signaling [27].…”

“…MMP-9 plays critical roles in regulation of EVT cell invasion, and an increased MMP-9 expression was found to be accompanied with the enhanced EVT cell migration [26]. More interestingly, it was reported that simultaneously decreased expression levels of MMP9 and RhoA were detected in trophoblast cells with reduced invasion ability and MMP9 could regulate trophoblast invasion ability through Rho-family signaling [27].…”

Chinese Herbal Medicines Have Potentially Beneficial Effects on the Perinatal Outcomes of Pregnant Women

“…(Tu Si Zi) are flavonoids, which can increase estrogen receptor expression in the hippocampus, hypothalamus, and pituitary as well as luteinizing hormone receptor expression in the ovaries ( Ke and Duan, 2013 ). Recent studies have reported that these flavonoids can promote the migration and invasion of extravillous trophoblast cells by increasing matrix metalloproteinase 9 (MMP9) expression and prevent miscarriage by activating Notch, AKT, and mitogen-activated protein kinase (MAPK) signaling pathways ( Gao et al, 2018 ). Previous research has shown that these herbs also nourish the kidneys and have similar effects to those of endocrine hormones by acting directly on the ovaries and regulating the hypothalamic-pituitary-ovarian axis to prevent miscarriage ( Zhu et al, 2014 ).…”

Ultra‐performance liquid chromatography quadrupole time‐of‐flight mass spectrometry–based plasma metabolomics study of hepatoprotective effect of cuscutae semen on CCl4‐induced liver injury model of rats

“…Shennong's Classic of Materia Medica recorded it as an upper-grade drug. Modern studies have indicated that CS contains flavonoids, polysaccharides, volatile oils, alkaloids, steroids, lignans, and so on; moreover, flavonoids and polysaccharides are the major bioactive components among them (Gao et al, 2018(Gao et al, , 2020Yang et al, 2017;Zhang et al, 2019). Currently, CS has demonstrated many pharmacological functions and biological activities, for instance, antioxidation effect (Lin et al, 2018;Szeto et al, 2012), anti-inflammatory effect (Liao et al, 2014), hepatoprotective activity (Yen et al, 2007(Yen et al, , 2008, and reversal of hormone levels (Yang et al, 2008).…”