Investigation of the Effect of High Glucose on Programmed Cell Death Mechanisms in Human Trophoblast Cells
Project no. 32225 · Status: Kapanmış · 2024
Summary
<p>Diabetes mellitus is a metabolic disease characterized by hyperglycemia and is becoming increasingly prevalent worldwide, including in women of reproductive age. The disease has 3 subclasses, Type 1, Type 2 and Gestational Type, with a prevalence of 7% in pregnant women, 86% of whom have Gestational Diabetes Mellitus (GDM). As a result of hyperglycemia caused by maternal and gestational diabetes, high glucose is transported from trophoblast cells to the fetus via glucose transporter proteins (GLUT), especially GLUT1 and GLUT3, and affects the development of placental and fetal cells. Diabetes often leads to complications in pregnancy, such as pre-eclampsia (PE) and fetal growth restriction (FGR), and these complications are associated with placental insufficiency. Placental insufficiency is thought to result from trophoblast cell dysfunction and high rates of cell death. While most studies to date have shown that apoptosis is the only programmed cell death mechanism involved in this process, recent studies suggest that necroptosis may also play a role. Apoptosis and necroptosis are programmed cell death mechanisms that share common upstream signaling elements and are initiated by activation of cell death receptors. As a result of activation and inhibition of important molecules such as cIAP and Casp8 in the signaling pathway, cells are directed towards survival, apoptosis or necroptosis. Apoptosis is mediated by caspases. Necroptosis is a process mediated by RIPK1, RIPK3 and MLKL. It is initiated when the RIPK1/RIPK3 complex forms in response to stimuli such as cytokines, pathogens or chemicals. Deacetylation of RIPK1 allows RIPK3 to phosphorylate MLKL, phosphorylated MLKL moves to the cell membrane and triggers necroptosis. There are studies linking necroptosis with neuropathy and retinopathy in diabetes. Although its role in diabetic pregnancy complications is not fully understood, studies in placentas of women with PE and FGR suggest that necroptosis is involved in the pathophysiology of these diseases. Understanding the effects of high glucose levels on programmed cell death mechanisms in trophoblast cells and the development of diabetes-related complications during pregnancy is important to guide future research and identify new possible therapeutic targets. In this study, it was aimed to investigate the effects of different glucose concentrations and high glucose on apoptosis and necroptosis, cell proliferation and migration from programmed cell death mechanisms in HTR8/SVneo human trophoblast cells. Considering human blood glucose levels for glucose concentrations in cell culture, low glucose group (1 mM), control group (5 mM) and high glucose groups (11 mM, 25 mM, 40 mM) were formed. MTT proliferation assay was performed to understand the effect of different glucose concentrations on cell viability. Annexin V/PI flow cytometry was used to examine apoptosis and necrosis rates. Caspase-3, Caspase-8, RIPK1, MLKL immunofluorescence staining (IF) was performed for protein level analysis of high glucose-induced apoptosis and necroptosis in trophoblast cells. Wound healing migration assay was performed to investigate cell migration capacity. Statistical analyses were performed and the results were evaluated. As a result, it was found that high glucose decreased viability and migration in HTR8/SVneo cells and induced apoptosis and necroptosis, which are programmed cell death mechanisms. In line with these findings, it is thought that possible new therapeutic targets may contribute to the treatment of diabetes-related pregnancy complications by targeting further research in placenta and trophoblast cells.</p><p><strong>Keywords:</strong> Apoptosis; Diabetes; Hyperglycemia; Necroptosis; Placenta; Trophoblast</p>