Open Access

CCR3‑shRNA promotes apoptosis and inhibits chemotaxis and degranulation of mouse mast cells

  • Authors:
    • Haisen Peng
    • Bing Liao
    • Xinhua Zhu
    • Yuehui Liu
    • Yinli Jiang
    • Shuhong Wu
  • View Affiliations

  • Published online on: May 12, 2020     https://doi.org/10.3892/etm.2020.8737
  • Pages: 1030-1038
  • Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Mast cells (MCs) are the major effector cells of allergic rhinitis (AR). The present study aimed to investigate the effects of C‑C chemokine receptor type 3 (CCR3) on the proliferation, apoptosis, chemotaxis and activated degranulation of mouse MCs. Mouse bone marrow‑derived MCs were cultured in vitro, purified and identified using toluidine blue staining and flow cytometry. Three different CCR3‑short hairpin (shRNA) lentiviral vectors were constructed and transfected into MCs, and the mRNA and protein expression levels of CCR3 were assessed by reverse transcription‑quantitative PCR and western blotting. Proliferation and apoptosis of the MCs were measured using Cell Counting kit‑8 (CCK‑8) assays and flow cytometry, respectively. MC chemotaxis was assessed by Transwell assay and quantified using flow cytometry. The activation of MC degranulation was examined using ELISAs. The results demonstrated that MCs were appropriately isolated, and identified that CCR3‑shRNA2 presented the higher knockdown effect among the three shRNAs tested. Following 96 h of transfection, the results of CCK‑8 and flow cytometry assays demonstrated that CCR3‑shRNA2 inhibited MC proliferation and promoted MC apoptosis. The results from the Transwell assay indicated that CCR3‑shRNA2 restrained MC chemotaxis, whereas ELISA results demonstrated that CCR3‑shRNA2 suppressed MC degranulation. In conclusion, CCR3‑shRNA2 effectively downregulated CCR3 mRNA and protein expression levels in mouse MCs. In addition, CCR3‑shRNA2 promoted MC apoptosis and suppressed the proliferation, chemotaxis and degranulation of mouse MCs, suggesting that CCR3‑shRNA2 may serve as a therapeutic tool for the treatment of allergic rhinitis.

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August-2020
Volume 20 Issue 2

Print ISSN: 1792-0981
Online ISSN:1792-1015

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APA
Peng, H., Liao, B., Zhu, X., Liu, Y., Jiang, Y., & Wu, S. (2020). CCR3‑shRNA promotes apoptosis and inhibits chemotaxis and degranulation of mouse mast cells. Experimental and Therapeutic Medicine, 20, 1030-1038. https://doi.org/10.3892/etm.2020.8737
MLA
Peng, H., Liao, B., Zhu, X., Liu, Y., Jiang, Y., Wu, S."CCR3‑shRNA promotes apoptosis and inhibits chemotaxis and degranulation of mouse mast cells". Experimental and Therapeutic Medicine 20.2 (2020): 1030-1038.
Chicago
Peng, H., Liao, B., Zhu, X., Liu, Y., Jiang, Y., Wu, S."CCR3‑shRNA promotes apoptosis and inhibits chemotaxis and degranulation of mouse mast cells". Experimental and Therapeutic Medicine 20, no. 2 (2020): 1030-1038. https://doi.org/10.3892/etm.2020.8737