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miR-21 Exerts Anti-proliferative and Pro-apoptotic Effects in LPS-induced WI-38 Cells via Directly Targeting TIMP3

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Abstract

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease, which was caused by a complex interplay of inflammatory responses and chronic damage. miR-21 is increased in patients with IPF, but its function in the embryonic lung-derived diploid fibroblasts cells subjected to LPS is elusive. miRNA expression profile was obtained from GEO database and target genes of miRNAs were forecasted by TargetScan. To mimic the LPS-induced injury, different concentrations of LPS were applied to treat WI-38 cells. Functional in vitro experiments were conducted to examine the role of miR-21 and TIMP3. Luciferase report assay was performed to verify the relationship between miR-21 and TIMP3. qRT-PCR, western blotting, and ELISA were conducted to detect the levels of the related miRNAs, proteins, and inflammatory factors. miR-21 presented higher levels in interstitial pneumonia patients and LPS-induced WI-38 cells. Overexpression of miR-21 was negatively correlated with the proliferative capability of LPS-treated WI-38 cells. miR-21 directly targets TIMP3. TIMP3 restored the suppressive impact of miR-21 mimic on the proliferation, while TIMP3 alleviated the promoting impact of miR-21 mimic on the apoptosis of WI-38 cells treated by LPS. miR-21 inhibited Bcl-2 but increased Bax, cleaved caspase-3, and cleaved caspase-9. Besides, miR-21 elevated the levels of IL-6 and IL-β but reduced the IL-10, which were weakened by TIMP3. Totally, miR-21 aggravated the LPS-induced lung injury and modulated inflammatory responses by targeting TIMP3.

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Authors and Affiliations

  1. Department of ICU, The Second People’s Hospital of Liaocheng, Linqing, Shandong, China

    Jin-Xiu Li & Feng-Yan Rong

  2. Department of ICU, Shandong First Medical University Affiliated Liaocheng Second Hospital, Linqing, Shandong, China

    Jin-Xiu Li & Feng-Yan Rong

  3. Department of ICU, Linqing People’s Hospital, Linqing, Shandong, China

    You Li

  4. Department of Pharmacy, The Second People’s Hospital of Liaocheng, Linqing, Shandong, China

    Tian Xia

  5. Department of Pharmacy, Shandong First Medical University Affiliated Liaocheng Second Hospital, Linqing, Shandong, China

    Tian Xia

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Li, JX., Li, Y., Xia, T. et al. miR-21 Exerts Anti-proliferative and Pro-apoptotic Effects in LPS-induced WI-38 Cells via Directly Targeting TIMP3. Cell Biochem Biophys 79, 781–790 (2021). https://doi.org/10.1007/s12013-021-00987-w

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