hsa-miR-1 and hsa-miR-1229 expression levels in recurrent miscarriage patients before and after lymphocyte therapy in comparison to control groups
Abstract
Recurrent pregnancy loss (RPL) occurs in 2-5% of women of childbearing age and is defined as 2 or more recurrent pregnancy failures before the 20th week. Immunological disorders, including an increase in the ratio of Th1/Th2 towards Th1 responses, an increase in the number of Th17 cells, a decrease in the number of Treg cells, and an imbalance between oxidative stress factors and antioxidants are known as the main causes of recurrent miscarriage. In the meantime, we will examine the amount of changes in the expression of two microRNAs, which has received less attention among the studies.
Methods: 25 patients with recurrent miscarriage and 25 healthy people with successful pregnancy were considered as the control group for this study. To prepare peripheral blood mononuclear cells (PBMCs) from peripheral blood, first 10 ml of heparinized blood is taken from patients and peripheral blood mononuclear cells are isolated using Ficol 1.077. According to the standard protocol, the number of 2 × 107 was injected subcutaneously to patients with recurrent miscarriage. The number of injections is three times with one-month intervals and Real-time PCR technique was used to check the expression level of hsa-miR-1 and hsa-miR-1229 molecules.
Findings: In this study, the gene expression level of micro-RNA 1 in RPL women before three cycles of subcutaneous injection of lymphocytes was 2.030±1.445 and the gene expression level of the same micro-RNA after the lymphocyte therapy of the mother was 1.101±0.4780. Also, the expression level of micro-RNA 1229 gene in these women was reported to be 2.100±0.6296 before treatment and 1.247±0.9631 after three cycles of treatment with paternal lymphocytes. This level of gene expression in women with a history of successful pregnancy as a control group, for micro-RNA 1 and micro-RNA 1229 genes is 1.000±0.08334 and 1.000±0.08091, respectively.