Document Type : Original Article
Authors
1
Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
2
Infertility Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
3
Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
4
Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
5
Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
6
Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
7
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
8
General Surgery, West Kazakhstan Marat Ospanov Medical University, Aktobe, Kazakhstan
9
Department of Oncology, West Kazakhstan Marat Ospanov Medical University, Aktobe, Kazakhstan
10
Department of Anatomy, School of Medical Sciences, Medicine, UNSW Sydney, PO Box 2052, Sydney, Australia
11
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
12
Reproductive Development, Murdoch Children's Research Institute, Melbourne, Victoria, Australia
13
Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
14
PerciaVista R&D Co. Shiraz, Iran
15
Department for Scientific Work, West Kazakhstan Marat Ospanov Medical University, Aktobe, Kazakhstan
10.22074/ijfs.2023.2007228.1496
Abstract
Background: This study aimed to investigate the effects of carob (Ceratonia siliqua L.) pod extract (CPE) on the viability of human endometrial mesenchymal stromal/stem cells (EnMSCs) and examine its impact on the mRNA and protein expression of DNA-methyltransferases (DNMT-1, DNMT-3A, and DNMT-3B), histone deacetylase-1 (HDAC-1), matrix metalloproteinase-2 (MMP-2), and cyclooxygenase-2 (COX-2) in endometriotic patients.
Materials and Methods: In an experimantal study, EnMSCs were derived from the endometrium of patients with endometrioma (OMA-EnMSCs) and deep infiltrative endometriosis samples of 10 women with endometriosis-associated infertility (E-EnMSCs), and were compared to EnMSCs derived from the endometrium of endometriosis-free, normal women as the control group (C-EnMSCs). The metabolic activity of the control and case groups was evaluated by treating them with different concentrations of CPE. Cell viability were analyzed using the MTT method. Real time RT-PCR and western blot assay were used to evaluate the expression of specific genes in mRNA and protein level, respectively.
Results: In the E-EnMSCs, treatment with 0.8 and 2 μg/mL of CPE resulted in the downregulation of COX-2 and HDAC-1 compared to the C-EnMSCs. Treatment with 0.8 μg/mL of CPE also decreased MMP-2 and DNMT-3B gene expression. Furthermore, COX-2 and DNMT-3A genes were significantly upregulated after treatment with 2 μg/mL of CPE. The expression of COX-2, HDAC-1, DNMT-1, DNMT-3A, and DNMT-3B peptides decreased in E-EnMSCs, OMA-EnMSCs, and C-EnMSCs after treatment with 0.8 and 2 μg/mL concentrations of CPE. GC analysis of CPE led to the identification of 14 bioactive compounds. Molecular docking showed the best position of each bioactive compounds on the different target proteins which are involved in the process of apoptosis in EnMSCs.
Conclusion: In vitro and in silico analysis of CPE bioactive compounds showed that they may downregulated the cell inflammatory pathway involved in the pathophysiology of endometriosis.
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