全反式维甲酸调节小鼠胚胎腭间质（MEPM）细胞的衰老。

IF 2.8 4区医学 Q2 REPRODUCTIVE BIOLOGY

Reproductive toxicology Pub Date : 2025-08-13 DOI:10.1016/j.reprotox.2025.109030

Yang Ma , Shiyi Huang , Yingshi Liang , Yuebin Zhu , Haofeng Huang , Bingna Zhang , Lungang Shi

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引用次数: 0

摘要

背景：非综合征性唇腭裂是最常见的先天性颅面畸形之一，其发病机制尚不清楚。多项相关研究证实，唇腭裂是环境和遗传因素共同作用的结果。视黄酸是维生素A的代谢产物之一，参与机体的多种生理功能，对调节细胞生长分化、维持人体正常发育至关重要。然而，全反式维甲酸（All-trans Retinoic Acid, atRA）的过量摄入是导致腭裂的病因之一。细胞衰老是近年来研究的热点和新方向，它与各种肿瘤和疾病的发生有关。本研究旨在从细胞衰老的角度，在小鼠胚胎腭间质（MEPM）细胞模型中验证atra诱导腭裂的作用机制。本研究旨在通过53/p21信号通路验证atra诱导腭裂的发病机制是否由于MEPM细胞的细胞衰老而发生。方法：取atra诱导腭裂腭组织进行切片染色和western blot检测。从腭组织中获得MEPM细胞，体外培养，然后用atRA处理MEPM细胞。验证atRA可通过p53/p21通路影响MEPM细胞的细胞增殖能力和细胞衰老，从而导致胚胎小鼠腭融合失败，导致腭裂。采用衰老相关β-半乳糖苷酶染色、CCK-8活性测定、细胞周期分析和western blot检测。结果：atra诱导腭裂组腭裂组织切片TUNEL细胞凋亡荧光染色无明显变化。然而，经atRA处理的MEPM细胞衰老增加，其特征是衰老相关β-半乳糖苷酶（SA-β-Gal）活性增强，细胞增殖减少，MEPM细胞周期停滞在G1期，衰老标志物p53和p21的表达增加。P53 /p21信号通路上调，可诱导细胞衰老，导致细胞增殖能力下降。结论：我们的实验结果表明，atRA可通过MEPM细胞的p53/p21信号通路增加细胞衰老，降低细胞活性和增殖能力，诱导细胞衰老的发生，导致胚胎小鼠腭裂。本研究旨在从细胞衰老的角度拓展atra诱发腭裂的作用机制，并为腭裂病因的研究提供新的思路。

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All-trans Retinoic Acid regulates cellular senescence of mouse embryonic palatal mesenchyme (MEPM) cells in developing cleft palates

Background

Non-syndromic cleft lip and palate is one of the most common congenital craniofacial malformations, however, its mechanism is not well understood. A number of relevant studies have confirmed that the cleft lip and palate is caused by the interaction of environmental and genetic factors. Retinoic acid is one of the metabolites of vitamin A and is involved in various physiological functions in the body, which is essential for the regulation of cell growth and differentiation and the maintenance of normal human development. However, excessive intake of All-trans Retinoic Acid (atRA) is one of the etiological factors contributing to the development of cleft palate. Cell senescence is a hot topic and a new direction in recent years, which is related to the occurrence of various tumors and diseases. From the perspective of cellular senescence, this study aimed to verify the mechanism of action of atRA-induced cleft palate in model of mouse embryonic palatal mesenchyme (MEPM) cells. This study aims to verify whether the pathogenesis of the atRA-induced cleft palate occurs because the cellular senescence of MEPM cells via 53/p21 signaling pathway.

Methods

The palate tissues of atRA-induced cleft palate were obtained for section staining and western blotting test. MEPM cells were obtained from palate tissues and cultured in vitro, and then MEPM cells in vitro were treated with atRA. To verify that the atRA could affect the cell proliferative ability and cellular senescence of MEPM cells through the p53/p21 pathway, which resulted in the failure of palatal fusion in embryonic mice, leading to cleft palate. The senescence-associated β-Galactosidase staining, CCK-8 activity assay, cell cycle analysis and western blotting test were used.

Results

In atRA-induced cleft palate group, tissue sections of the palate showed that there was no change in TUNEL apoptosis fluorescence staining. However, there was increased cellular senescence in MEPM cells treated with atRA as characterized by enhancing senescence-associated β-galactosidase (SA-β-Gal) activity, reducing cell proliferation, inducing MEPM cells cell cycle arrest at G1 phase and increasing expression of the senescence markers p53 and p21. p53/p21 signaling pathway was up-regulated, which could induce the cells to undergo senescence, resulting in a decrease of cell proliferative ability.

Conclusions

Our experimental results have shown that atRA could increase cell senescence through the p53/p21 signaling pathway in MEPM cells and diminish cell activity and proliferation ability, inducing the occurrence of cellular senescence and resulting in cleft palate in the embryonic mouse. From the viewpoint of cellular senescence, this study is intended to expand the mechanism of action of atRA-induced cleft palate as well as to provide new ideas for the study of the etiology of cleft palate.

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来源期刊

Reproductive toxicology 生物-毒理学

CiteScore

6.50

自引率

3.00%

发文量

131

审稿时长

45 days

期刊介绍： Drawing from a large number of disciplines, Reproductive Toxicology publishes timely, original research on the influence of chemical and physical agents on reproduction. Written by and for obstetricians, pediatricians, embryologists, teratologists, geneticists, toxicologists, andrologists, and others interested in detecting potential reproductive hazards, the journal is a forum for communication among researchers and practitioners. Articles focus on the application of in vitro, animal and clinical research to the practice of clinical medicine. All aspects of reproduction are within the scope of Reproductive Toxicology, including the formation and maturation of male and female gametes, sexual function, the events surrounding the fusion of gametes and the development of the fertilized ovum, nourishment and transport of the conceptus within the genital tract, implantation, embryogenesis, intrauterine growth, placentation and placental function, parturition, lactation and neonatal survival. Adverse reproductive effects in males will be considered as significant as adverse effects occurring in females. To provide a balanced presentation of approaches, equal emphasis will be given to clinical and animal or in vitro work. Typical end points that will be studied by contributors include infertility, sexual dysfunction, spontaneous abortion, malformations, abnormal histogenesis, stillbirth, intrauterine growth retardation, prematurity, behavioral abnormalities, and perinatal mortality.