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Integrated miRNA-seq analysis reveals the molecular mechanism underlying the effect of acupuncture on endometrial receptivity in patients undergoing fertilization: embryo transplantation

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Abstract

Endometrial receptivity (ER) is the main factor affecting in vitro fertilization (IVF) and embryo transplantation. Previous studies have shown that acupuncture might be useful for increasing ER. However, the underlying microRNA (miRNA) molecular mechanisms deserve to be further elucidated. In this study, we performed small RNA sequencing of endometrial samples from infertile women who either underwent acupuncture therapy or did not. Differentially expressed microRNAs (DEmiRNAs) were identified and used to predict target genes. Then, the functional and pathway were analyzed for the target genes. Moreover, quantitative reverse transcription PCR (qRT-PCR) was performed to validate the RNA-seq results. Finally, the miRNA-gene network was conducted by Cytoscape. A total of 39 DEmiRNAs were identified between the acupuncture group and the control group. The functional enrichment analysis suggested that the target genes of the DEmiRNAs were significantly enriched in GO biological process (BP) terms associated with transcription, such as regulation of DNA-templated transcription and positive regulation of DNA-templated transcription. The pathway analysis showed that DEmiRNAs might be involved in acupuncture therapy via Endocytosis, Axon guidance, Oxytocin signaling, the Hippo pathway, and Estrogen signaling pathways. Significant downregulation of hsa-miR-449a and hsa-miR-449b-3p, and significant upregulation of hsa-miR-3135b and hsa-miR-345-3p in the RNA-seq results were validated by qRT-PCR. Besides, these four DEmiRNAs and their 34 target genes conducted a miRNA–gene network. Our results predict that hsa-miR-449a, hsa-miR-3135b and hsa-miR-345-3p may underly mechanisms by which acupuncture therapy help increase ER and promote endometrium receptivity in preparation for in vitro fertilization and embryo transplantation.

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Data availability

The raw data were uploaded to Sequence Read Archive (SRA) (http://www.ncbi.nlm.nih.gov/) with accession PRJNA587020.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China [Grant numbers 81473767, 81873371, 81403477, 81603674, 81403481, 81804179] and by the Jiangsu Provincial Science and Technology Project of the Traditional Chinese Medicine Bureau [Grant number YB2017005].

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Author notes

  1. Yanyun Mu and Qian Li contributed equally to this work and should be considered co-first authors.

Authors and Affiliations

  1. College of Acupuncture and Tuina, Nanjing University of Chinese Medicine, Nanjing, 210023, Jiangsu, China

    Yanyun Mu, Qian Li, Jie Cheng, Jie Shen, Xun Jin, Youling Gao & Youbing Xia

  2. Preventive Treatment Center, Nanjing Hospital of Traditional Chinese Medicine, Nanjing, 210002, Jiangsu, China

    Zhengyun Xie

  3. Pain Management, Zhongda Hospital Southest University, Nanjing, 210009, Jiangsu, China

    Zhao Gao

  4. Beijing Haidian Garrison District Fifth Retired Cadre Sanatorium, Beijing, 100091, China

    Wenjing Zhang

  5. Acupuncture and Moxibustion Department, Nanjing Hospital of Traditional Chinese Medicine, Preventive Treatment Center, Nanjing, 210002, Jiangsu, China

    Qixin Hua

  6. College of Medicine and Life Sciences, Nanjing University of Chinese Medicine, Nanjing, 210023, Jiangsu, China

    Liangjun Xia

  7. The Affiliated Hospital, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, China

    Youbing Xia

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  1. Yanyun Mu
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Contributions

Conceptualization: Youbing Xia; Methodology: Yanyun Mu, Qian Li, Jie Cheng; Formal analysis and investigation: Yanyun Mu, Qian Li, Jie Shen, Zhengyun Xie, Zhao Gao, Wenjing Zhang, Qixin Hua; Writing—original draft preparation: Yanyun Mu, Jie Shen, Youling Gao; Writing—review and editing: Qian Li, Xun Jin, Liangjun Xia; Funding acquisition: Youbing Xia; Resources: Jie Cheng, …; Supervision: Youbing Xia. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Youbing Xia.

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The authors declare that there is no conflict of interest regarding the publication of this paper.

Ethical approval

This study was approved by the ethics committee of Nanjing Medical University (No. 2014, 204).

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All women provided written informed consent.

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Mu, Y., Li, Q., Cheng, J. et al. Integrated miRNA-seq analysis reveals the molecular mechanism underlying the effect of acupuncture on endometrial receptivity in patients undergoing fertilization: embryo transplantation. 3 Biotech 10, 6 (2020). https://doi.org/10.1007/s13205-019-1990-3

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