The Effect of Endometrial Cell Culture on alpha 3 and beta 1 integrin Genes and Protein Expression in Type 2 Diabetic Rats at The Time of Implantation

Abstract

Objective: Given the prevalence of fertility problems in couples and the defect in embryo implantation as well as the low success rate of assisted reproductive techniques, it is necessary to investigate the causes of this phenomenon. Type 2 diabetes mellitus (T2DM) is a metabolic disease with multiple effects on various organs as well as the endometrium. In this study, the effects of endometrial cell culture on the expression of alpha 3 and beta 1 integrin genes and protein in type 2 diabetic rats were investigated. Materials and Methods: In this experimental study, 35 female rats were divided into five groups: control, sham, diabetic, Pioglitazone-treated and Metformin-treated groups. First, rats were maintained in diabetic condition for 4 weeks. Then, treatment was performed for the next four weeks. Four weeks after induction of diabetes, rats were sacrificed at the time of embryo implantation. The uterus was removed. Endometrial cells were isolated and cultured for 7 days. Immunocytochemistry staining was used to confirm endometrial cells. Expression of alpha 3 and beta 1 integrin genes was determined by real-time polymerase chain reaction (PCR) technique and the alpha 3 beta 1 protein content measured using Western blot both before and after endometrial cell culture. Result: The expression level of alpha 3 integrin gene in the Pioglitazone-treated group compared with metformin-treated group was significantly decreased (P<0.001). The same result was observed in beta 1 integrin gene expression (P=0.004). Also, the alpha 3 beta 1 protein level increased in all diabetic groups, but its reduction was significantly greater in pioglitazone-treated group (P=0.004) Conclusion: T2DM altered the expression of alpha 3 and beta 1 integrin genes and related proteins, which endometrial cell culture regulated this disorder. According to these results, may be the endometrial cell culture can reduce the adverse effects of diabetes on alpha 3 and beta 1 integrin expression at the level of gene and protein, in endometrial cells.