Role of ALKBH5-mediated m6A modification in endometrial decidualization of mice in early pregnancy

He Sihui; Li Weike; Gao Rufei; Chen Xuemei; Long Jing; Chen Chenxi; Dai Dexian; Wang Yingxiong; Li Na

doi:10.13406/j.cnki.cyxb.003949

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Role of ALKBH5-mediated m6A modification in endometrial decidualization of mice in early pregnancy

Basic research | 更新时间：2025-11-20

- Role of ALKBH5-mediated m6A modification in endometrial decidualization of mice in early pregnancy
- Journal of Chongqing Medical University Vol. 50, Issue 11, Pages: 1541-1549(2025)
- 作者机构：
  
  重庆医科大学公共卫生学院生殖与发育教育部国际合作联合实验室，重庆 400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.003949
  CLC： R114
- Received：24 June 2025，
  
  Revised：2025-08-02，
  
  Accepted：15 September 2025，
  
  Published：28 November 2025
- 稿件说明：
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贺思慧,李伟轲,高茹菲等.ALKBH5介导的m6A修饰在早孕小鼠子宫内膜蜕膜化中的作用研究[J].重庆医科大学学报,2025,50(11):1541-1549.

He Sihui,Li Weike,Gao Rufei,et al.Role of ALKBH5-mediated m6A modification in endometrial decidualization of mice in early pregnancy[J].Journal of Chongqing Medical University,2025,50(11):1541-1549.
贺思慧,李伟轲,高茹菲等.ALKBH5介导的m6A修饰在早孕小鼠子宫内膜蜕膜化中的作用研究[J].重庆医科大学学报,2025,50(11):1541-1549. DOI： 10.13406/j.cnki.cyxb.003949.

He Sihui,Li Weike,Gao Rufei,et al.Role of ALKBH5-mediated m6A modification in endometrial decidualization of mice in early pregnancy[J].Journal of Chongqing Medical University,2025,50(11):1541-1549. DOI： 10.13406/j.cnki.cyxb.003949.

摘要

目的

探讨烷基化修复同源物5（alkylation repair homolog 5，ALKBH5）介导的N6-甲基腺苷（N6-methyladenosine，m6A）修饰在子宫内膜蜕膜化过程中的作用。

方法

构建妊娠小鼠模型和假孕小鼠模型，通过实时荧光定量PCR、蛋白质印迹法检测ALKBH5在子宫内膜中的表达规律。构建人工诱导蜕膜化的小鼠模型及细胞模型，采用实时荧光定量PCR、蛋白质印迹法、免疫组织化学法检测蜕膜化相关标志物表达。采用EpiQuik m6A RNA甲基化定量试剂盒检测m6A水平。构建干扰ALKBH5表达的人工诱导蜕膜化小鼠模型及细胞模型，利用实时荧光定量PCR、蛋白质印迹法、免疫组织化学法检测蜕膜化相关标志物、细胞增殖标志分子和凋亡分子的表达。采用流式细胞分析术检测细胞凋亡率。

结果

在妊娠小鼠模型中，子宫胚胎着床点ALKBH5的表达明显高于着床旁；在人工诱导蜕膜化的小鼠和细胞模型中，与对照组相比，诱导组ALKBH5的表达水平明显升高，同时m6A水平明显降低。若干扰ALKBH5表达，则可导致m6A水平升高，进而抑制基质细胞增殖，诱导细胞凋亡，最终损害子宫内膜蜕膜化的正常进程。

结论

ALKBH5缺失可导致子宫内膜m6A水平升高，从而导致蜕膜化损伤，为m6A修饰在蜕膜化的研究奠定了基础。

Abstract

Objective

To investigate the role of alkylation repair homolog 5（ALKBH5）-mediated N6-methyladenosine （m6A） modification in endometrial decidualization.

Methods

The mouse models of pregnancy and pseudopregnancy were established，and quantitative real-time PCR and Western blot were used to measure the expression pattern of ALKBH5 in the endometrium. The mouse and cell models of artificially induced decidualization were established，and quantitative real-time PCR，Western blot，and immunohistochemistry were used to measure the expression levels of decidualization-related markers. The EpiQuik m6A RNA methylation quantification kit was used to measure the level of m6A. The mouse and cell models of artificially induced decidualization with interference of ALKBH5 expression were established，and quantitative real-time PCR，Western blot，and immunohistochemistry were used to measure the expression levels of decidualization-related markers，cell proliferation marker molecules，and apoptosis molecules. Flow cytometry was used to measure cell apoptosis rate.

Results

In the mouse model of pregnancy，the expression level of ALKBH5 at the uterine embryo implantation site was significantly higher than that adjacent to the implantation site，and in the mouse and cell models of artificially induced decidualization，compared with the control group，the induction group had a significant increase in the expression level of ALKBH5 and a significant reduction in the level of m6A. Inhibiting the expression of ALKBH5 led to an increase in the level of m6A，which in turn inhibited the proliferation of stromal cells，induced cell apoptosis，and ultimately impaired the normal process of endometrial decidualization.

Conclusion

ALKBH5 deficiency leads to an increase in the level of m6A and decidualization injury in the endometrium，which lays a foundation for the research on m6A modification in decidualization.

关键词

Keywords

references

Makrigiannakis A ， Minas V . Mechanisms of implantation . Reprod Biomed Online . 2007 ， 14 （ 1 ）： 102 - 109

Cha J ， Sun XF ， Dey SK . Mechanisms of implantation：strategies for successful pregnancy [J ] . Nat Med ， 2012 ， 18 （ 12 ）： 1754 - 1767 ．

Wang J ， Gao FC ， Zhao XH ， et al . Integrated analysis of the transcriptome-wide m6A methylome in preeclampsia and healthy control placentas [J ] . PeerJ ， 2020 ， 8 ： e9880 ．

Long J ， Li WK ， Chen MY ， et al . Uterine deficiency of Dnmt3b impairs decidualization and causes consequent embryo implantation defects [J ] . Cell Biol Toxicol ， 2023 ， 39 （ 3 ）： 1077 - 1098 ．

Liang XH ， Deng WB ， Liu YF ， et al . Non-coding RNA LINC00473 mediates decidualization of human endometrial stromal cells in response to cAMP signaling [J ] . Sci Rep ， 2016 ， 6 ： 22744 ．

Zhao BS ， Roundtree IA ， He C . Post-transcriptional gene regulation by mRNA modifications [J ] . Nat Rev Mol Cell Biol ， 2017 ， 18 （ 1 ）： 31 - 42 ．

Zhou YJ ， Kong Y ， Fan WG ， et al . Principles of RNA methylation and their implications for biology and medicine [J ] . Biomed Pharmacother ， 2020 ， 131 ： 110731 ．

He PC ， He C . M（6） A RNA methylation：from mechanisms to therapeutic potential [J ] . EMBO J ， 2021 ， 40 （ 3 ）： e105977 ．

Tan XS ， Zheng CH ， Zhuang YH ， et al . The m6A reader PRRC2A is essential for meiosis I completion during spermatogenesis [J ] . Nat Commun ， 2023 ， 14 （ 1 ）： 1636 ．

Ivanova I ， Much C ， Di Giacomo M ， et al . The RNA m6A reader YTHDF2 is essential for the post-transcriptional regulation of the maternal transcriptome and oocyte competence [J ] . Mol Cell ， 2017 ， 67 （ 6 ）： 1059 - 1067 .

Hua WF ， Zhao Y ， Jin XH ， et al . METTL3 promotes ovarian carcinoma growth and invasion through the regulation of AXL translation and epithelial to mesenchymal transition [J ] . Gynecol Oncol ， 2018 ， 151 （ 2 ）： 356 - 365 ．

Garrido-Gómez T ， Castillo-Marco N ， Cordero T ， et al . Decidualization resistance in the origin of preeclampsia [J ] . Am J Obstet Gynecol ， 2022 ， 226 （ 2S ）： S886 - S894 ．

Liu L ， Li H ， Hu DY ， et al . Insights into N6-methyladenosine and programmed cell death in cancer [J ] . Mol Cancer ， 2022 ， 21 （ 1 ）： 32 ．

Chen YS ， Ouyang XP ， Yu XH ， et al . N6-adenosine methylation （m（6）A） RNA modification：an emerging role in cardiovascular diseases [J ] . J Cardiovasc Transl Res ， 2021 ， 14 （ 5 ）： 857 - 872 ．

Liao ZW ， Wang J ， Xu MR ， et al . The role of RNA m6A demethylase ALKBH5 in the mechanisms of fibrosis [J ] . Front Cell Dev Biol ， 2024 ， 12 ： 1447135 ．

Ning YC ， Chen J ， Shi YQ ， et al . Genistein ameliorates renal fibrosis through regulation snail via m6A RNA demethylase ALKBH5 [J ] . Front Pharmacol ， 2020 ， 11 ： 579265 ．

Li Y ， Li MF ， Liu J ， et al . Altered m6A modification is involved YAP-mediated apoptosis response in 4-vinylcyclohexene diepoxide induced ovotoxicity [J ] . Ecotoxicol Environ Saf ， 2023 ， 262 ： 115192 ．

Bai L ， Xiang Y ， Tang MY ， et al . ALKBH5 controls the meiosis-coupled mRNA clearance in oocytes by removing the N （6）-methyladenosine methylation [J ] . Nat Commun ， 2023 ， 14 （ 1 ）： 6532 ．

Zhang XS ， Deng SB ， Peng Y ， et al . ALKBH5 inhibits TNF-α-induced apoptosis of HUVECs through Bcl-2 pathway [J ] . Open Med ， 2022 ， 17 （ 1 ）： 1092 - 1099 ．

Tao M ， Li XY ， He L ， et al . Decreased RNA m（6）a methylation enhances the process of the epithelial mesenchymal transition and vasculogenic mimicry in glioblastoma [J ] . Am J Cancer Res ， 2022 ， 12 （ 2 ）： 893 - 906 ．

Han ZB ， Wang XX ， Xu ZH ， et al . ALKBH5 regulates cardiomyocyte proliferation and heart regeneration by demethylating the mRNA of YTHDF1 [J ] . Theranostics ， 2021 ， 11 （ 6 ）： 3000 - 3016 ．

Zhu ZY ， Qian Q ， Zhao X ， et al . N（6）-methyladenosine ALKBH5 promotes non-small cell lung cancer progress by regulating TIMP3 stability [J ] . Gene ， 2020 ， 731 ： 144348 ．

Fonseca BM ， Correia-da-Silva G ， Teixeira NA . The endocannabinoid anandamide induces apoptosis of rat decidual cells through a mechanism involving ceramide synthesis and p38 MAPK activation [J ] . Apoptosis ， 2013 ， 18 （ 12 ）： 1526 - 1535 ．

Zhao SS ， Lu JY ， Chen YX ， et al . Exploration of the potential roles of m6A regulators in the uterus in pregnancy and infertility [J ] . J Reprod Immunol ， 2021 ， 146 ： 103341 ．

Helm M ， Motorin Y . Detecting RNA modifications in the epitranscriptome：predict and validate [J ] . Nat Rev Genet ， 2017 ， 18 （ 5 ）： 275 - 291 .

Ng SW ， Norwitz GA ， Pavlicev M ， et al . Endometrial decidualization：the primary driver of pregnancy health [J ] . Int J Mol Sci ， 2020 ， 21 （ 11 ）： 4092 .

Li XC ， Jin F ， Wang BY ， et al . The m6A demethylase ALKBH5 controls trophoblast invasion at the maternal-fetal interface by regulating the stability of CYR61 mRNA [J ] . Theranostics ， 2019 ， 9 （ 13 ）： 3853 - 3865 ．

Shi HL ， Wei JB ， He C . Where，when，and how：context-dependent functions of RNA methylation writers，readers，and erasers [J ] . Mol Cell ， 2019 ， 74 （ 4 ）： 640 - 650 ．

Qu JW ， Hou YF ， Chen QX ， et al . RNA demethylase ALKBH5 promotes tumorigenesis in multiple myeloma via TRAF1-mediated activation of NF-κB and MAPK signaling pathways [J ] . Oncogene ， 2022 ， 41 （ 3 ）： 400 - 413 ．

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