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Integrative Analysis of MicroRNAs and mRNAs in LPS-Induced Macrophage Inflammation Based on Adipose Tissue Stem Cell Therapy

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Abstract

Severe inflammation can lead to multiple organ dysfunction syndrome, which has high mortality. Adipose-derived stem cells have been shown to affect the inflammatory response of macrophages. However, the molecular mechanism of the anti-inflammatory capacity of adipose-derived stem cells (ADSCs) remains to be understood. In the present study, a macrophage inflammation model was established by LPS, and treated with different volumes of ADSC supernatant. Then, we investigated the key genes in the LPS group and treatment group by RT-PCR, RNA sequencing technology, and bioinformatics analysis. A total of 26 miRNAs and 11,882 mRNAs were differentially expressed between them. The expression of 15 of the miRNAs (9 upregulated and 6 downregulated) was confirmed by RT-PCR. GO and KEGG pathway analyses of the targets of the 9 significantly upregulated miRNAs showed that they were related to immune system process, inflammatory response, lipopolysaccharide, and TNF-α, NF-κB, Toll-like receptor, and MAPK signaling pathways. Moreover, a miRNA-mRNA network also revealed 8 important genes (Mapkapk2, Sepp1, Cers6, Snn, ZfP568, Ccdc93, Pofut1, Pik3cd). We finally confirmed the expression of these 8 targeted genes by performing the RT-PCR analysis. This study may provide a new understanding of the molecular mechanism of ADSCs in the inflammatory response related to multiple miRNAs and mRNAs.

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Data Availability

All data generated and/or analyzed during this study are included in this published article.

Abbreviations

ADSCs:

adipose tissue–derived stem cells

ADSC-sup:

adipose tissue–derived stem cell supernatant

BMDM:

bone marrow–derived macrophage

BSA:

albumin bovine serum

DE:

differentially expressed

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

fetal bovine serum

IL:

interleukin

LPS:

lipopolysaccharide

MSC:

mesenchymal stem cell

PBS:

phosphate-buffered saline

RNA-seq:

RNA sequencing

RT-PCR:

reverse transcription polymerase chain reaction

TNF:

tumor necrosis factor

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Funding

This work was supported by the National Natural Science Foundation of China (81501663 and 81530064).

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Author notes

  1. Xiaozhi Bai, Ting He and Mingchuan Liu contributed equally to this work.

Authors and Affiliations

  1. Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, No. 127 West Changle Road, Xi’an, 710032, Shaanxi, China

    Xiaozhi Bai, Ting He, Jie Chen, Yang Liu, Chen Yang, Wenbin Jia, Ke Tao, Juntao Han & Dahai Hu

  2. Brigade 4, College of Basic Medicine, Fourth Military Medical University, No. 169 West Changle Road, Xi’an, 710032, Shaanxi, China

    Mingchuan Liu & Lincheng Li

  3. Chinese People’s Liberation Army Hospital 961, No. 71 Youzheng Road, Qiqihar, 161000, Heilongjiang, China

    Mengyuan Cao

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  1. Xiaozhi Bai
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Contributions

XB, TH, and ML contribute equally as first authors. XB and DH designed the project. ML, LL, MC, and YL performed most of the experiments. JC and ML analyzed data. CY verified cells and built the macrophage inflammation model. CY, TK, and WJ contributed to results’ analysis. XB, JH, and DH revised the manuscript and approved the final submission.

Corresponding authors

Correspondence to Juntao Han or Dahai Hu.

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This study was carried out in accordance with the principles of ARRIVE approved by the Ethics Committee of Xijing Hospital, affiliated with the Fourth Military Medical University (No.: XJYYLL-2015206).

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Bai, X., He, T., Liu, M. et al. Integrative Analysis of MicroRNAs and mRNAs in LPS-Induced Macrophage Inflammation Based on Adipose Tissue Stem Cell Therapy. Inflammation 44, 407–420 (2021). https://doi.org/10.1007/s10753-020-01345-3

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