Integrative Analysis of MicroRNAs and mRNAs in LPS-Induced Macrophage Inflammation Based on Adipose Tissue Stem Cell The
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ORIGINAL ARTICLE
Integrative Analysis of MicroRNAs and mRNAs in LPS-Induced Macrophage Inflammation Based on Adipose Tissue Stem Cell Therapy Xiaozhi Bai,1 Ting He,1 Mingchuan Liu,2 Lincheng Li,2 Jie Chen,1 Mengyuan Cao,3 Yang Liu,1 Chen Yang,1 Wenbin Jia,1 Ke Tao,1 Juntao Han,1,4 and Dahai Hu 1,4 (Received March 13, 2020; accepted September 10, 2020)
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 RTPCR, 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. KEY WORDS: inflammation; miRNAs; macrophages; adipose tissue stem cells; RNA sequencing.
Xiaozhi Bai, Ting He and Mingchuan Liu contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10753-020-01345-3) contains supplementary material, which is available to authorized users. 1
Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, No. 127 West Changle Road, Xi’an, 710032, Shaanxi, China
2
Brigade 4, College of Basic Medicine, Fourth Military Medical University, No. 169 West Changle Road, Xi’an, 710032, Shaanxi, China 3 Chinese People’s Liberation Army Hospital 961, No. 71 Youzheng Road, Qiqihar, 161000, Heilongjiang, China 4 To whom correspondence should be addressed at Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, No. 127 West Changle Road, Xi’an, 710032, Shaanxi, China. E-mails: [email protected]; [email protected]
0360-3997/20/0000-0001/0 # 2020 Springer Science+Business Media, LLC, part of Springer Nature
Bai, He, Liu, Li, Chen, Cao, Liu, Yang, Jia, Tao, Han, and Hu INTRODUCTION Inflammation is an essential immune response to infection and tissue injury. However, excessive inflammation may lead to undesirable conseque
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