Ceratonia siliqua L. pod Effects on Viability Gene Expression of Endometrial Mesenchymal Stromal/Stem Cells Isolated from Women with Endometriosis-Associated Infertility.

IF 2.3 Q2 OBSTETRICS & GYNECOLOGY

International Journal of Fertility & Sterility Pub Date : 2024-10-30 DOI:10.22074/ijfs.2023.2007228.1496

Zahra Khodabandeh, Bahia Namavar Jahromi, Atefe Hashemi, Kamran Hessami, Iman Jamhiri, Shahrokh Zare, Parmis Badr, Aida Iraji, Tahere Poordast, Neda Baghban, Arezoo Khoradmehr, Nadiar Maratovich Mussin, Asset Askerovich Kaliyev, Yerbolat Maratovich Iztleuov, Reza Shirazi, Mahdi Mahdipour, Shabnam Bakhshalizadeh, Farhad Rahmanifar, Nazanin Jafari, Nader Tanideh, Amin Tamadon

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Abstract

Background: This study aims to investigate the effects of carob (Ceratonia siliqua L.) pod extract (CPE) on the viability of human endometrial mesenchymal stromal/stem cells (EnMSCs) and its impact on mRNA and protein expressions of DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B), histone deacetylase 1 (HDAC1), matrix metalloproteinase-2 (MMP2), and cyclooxygenase-2 (COX-2) in endometriotic patients.

Materials and methods: In this experimental study, EnMSCs were derived from endometrium of patients with ovarian endometrioma (OMA-EnMSCs group) and deep infiltrative endometriosis (DIE) samples of 10 endometriosisassociated infertility (EAI) women (E-EnMSCs group) and compared to EnMSCs derived from the endometrium of an endometriosis-free, normal woman as the control group (C-EnMSCs). The metabolic activity of the control and case groups were evaluated by treating them with different concentrations of CPE. Cell viability was analysed by MTT. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to evaluate the expression of specific genes at the mRNA and protein levels, respectively.

Results: Treatment with 0.8 and 2 μg/mL of CPE downregulated COX-2 and HDAC1 in the E-EnMSC group compared to the C-EnMSCs group. Treatment with 0.8 μg/mL of CPE also decreased MMP2 and DNMT3B gene expressions. The COX-2 and DNMT3A genes were significantly upregulated after treatment with 2 μg/mL of CPE. Expressions of the COX-2, HDAC1, DNMT1, DNMT3A, and DNMT3B peptides decreased in the all three groups after treatment with 0.8 and 2 μg/mL of CPE. Gas chromatography-mass spectroscopy (GC-MS) analysis of CPE identified 14 bioactive compounds. Molecular docking showed the best position of each bioactive compound on the different target proteins that are involved in the process of apoptosis in EnMSCs.

Conclusion: In vitro and in silico analyses of CPE bioactive compounds show that they may downregulate the cell inflammatory pathway involved in the pathophysiology of endometriosis.

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Ceratonia siliqua L. pod 对从子宫内膜异位症相关不孕症妇女体内分离的子宫内膜间质基质/干细胞活力基因表达的影响。

背景：本研究旨在探讨角豆树（Ceratonia siliqua L.）荚果提取物（CPE）对子宫内膜异位症患者的人子宫内膜间充质基质/干细胞（EnMSCs）活力的影响及其对 DNA 甲基转移酶（DNMT1、DNMT3A 和 DNMT3B）、组蛋白去乙酰化酶 1（HDAC1）、基质金属蛋白酶-2（MMP2）和环氧化酶-2（COX-2）的 mRNA 和蛋白表达的影响：在这项实验研究中，EnMSCs来源于卵巢子宫内膜瘤患者的子宫内膜（OMA-EnMSCs组）和10名子宫内膜异位症相关不孕症（EAI）妇女的深部浸润性子宫内膜异位症（DIE）样本（E-EnMSCs组），并与来源于无子宫内膜异位症的正常妇女子宫内膜的EnMSCs作为对照组（C-EnMSCs）进行比较。通过使用不同浓度的 CPE 对对照组和病例组的代谢活性进行评估。用 MTT 分析细胞活力。实时逆转录聚合酶链反应（RT-PCR）和 Western 印迹分别用于评估特定基因在 mRNA 和蛋白质水平上的表达：结果：与C-EnMSCs组相比，用0.8和2 μg/mL的CPE处理可下调E-EnMSCs组的COX-2和HDAC1。0.8 μg/mL的CPE还能降低MMP2和DNMT3B基因的表达。经 2 μg/mL CPE 处理后，COX-2 和 DNMT3A 基因表达明显上调。经 0.8 和 2 μg/mL CPE 处理后，三组中 COX-2、HDAC1、DNMT1、DNMT3A 和 DNMT3B 肽的表达均有所下降。CPE 的气相色谱-质谱（GC-MS）分析确定了 14 种生物活性化合物。分子对接显示了每种生物活性化合物在参与 EnMSCs 细胞凋亡过程的不同靶蛋白上的最佳位置：对 CPE 生物活性化合物进行的体外和硅学分析表明，它们可能会降低参与子宫内膜异位症病理生理学的细胞炎症通路。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Fertility & Sterility 医学-妇产科学

CiteScore

4.20

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： International Journal of Fertility & Sterility is a quarterly English publication of Royan Institute . The aim of the journal is to disseminate information through publishing the most recent scientific research studies on Fertility and Sterility and other related topics. Int J Fertil Steril has been certified by Ministry of Culture and Islamic Guidance in 2007 and was accredited as a scientific and research journal by HBI (Health and Biomedical Information) Journal Accreditation Commission in 2008. Int J Fertil Steril is an Open Access journal.