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Silencing TLR4/MyD88/NF-κB Signaling Pathway Alleviated Inflammation of Corneal Epithelial Cells Infected by ISE

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Abstract

The regulatory role of toll-like receptor 4 (TLR4) in the inactivate staphylococcus epidermidis (ISE)-induced cornea inflammation is not well investigated. Here, TLR4 silence could decrease inflammatory cytokines in corneal epithelial cells treated with ISE. The mouse corneal epithelial cells were exposed to ISE for 24 h, either alone or with the NF-κB inhibitor, TLR4 lentivirus to bilaterally (knock-down or and overexpression). The expression of TLR4 in mouse corneal epithelial cells was investigated using western blot and qRT-PCR assay. The inflammatory cytokine levels were evaluated by qRT-PCR and ELISA, respectively. The relative impact factors of TLR4-mediated NF-κB signaling detected using western blot assay. Results show the expression levels of TLR4 and some inflammatory cytokines were significantly increased in corneal epithelial cells treated with ISE. TLR4 Silence markedly decreased ISE-induced production of IL12, TNF-α, CCL5, and CCL9 in corneal epithelial cells. Furthermore, the nuclear translocation of NF-κB p65 and myeloid differentiation protein 88 (MyD88) in the cells treated with ISE were further reduced by silencing TLR4. Inhibition of TLR4-mediated NF-κB signaling by using BAY11-7082 also alleviated ISE-induced inflammation. In the rescue experiment, transfected the stable TLR4 silenced corneal epithelial cells with TLR4 overexpression lentivirus, we found that TLR4 overexpression can restore the down-regulation of TLR4 and inflammatory cytokines (IL12, TNF-α, CCL9) caused by TLR4 knocked down. Therefore, ISE-induced cornea inflammation was due to the activation of the TLR4/MyD88/NF-κB signaling pathway, and dramatically stimulated IL12, TNF-α, CCL9 secretion. TLR4 silence presented mitigates damage in corneal epithelial cells treated with ISE.

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Funding

This study was supported by grants from the Natural Science Foundation of Jiangsu Province, China (15KJB180015), Nantong Science and Technology Program (JC2018090), and Scientific Innovation Research of College Graduates in Jiangsu Province (KYCX18-2415).

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  1. Liucheng Wu and Lili Du contributed equally to this work.

Authors and Affiliations

  1. Laboratory Animal Center, Nantong University, Nantong, 226001, China

    Liucheng Wu, Lili Du, Zhiheng Chen, Yu Ma, Guiqing Ji, Yixiang Shao & Xiaoqing Peng

  2. Department of Thoracic Surgery, Affiliated Hospital of Nantong University, Nantong, 226001, China

    Qianqian Ju

  3. School of Nursing, Nantong University, Nantong, 226001, China

    Ting Bai

  4. Pharmacy Department, Nantong Hospital of Traditional Chinese Medicine, Nantong, 226001, China

    Yu Wu

  5. School of Pharmacy, Nantong University, Nantong, 226001, China

    Zhaoguo Liu

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  1. Liucheng Wu
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Correspondence to Xiaoqing Peng.

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Wu, L., Du, L., Ju, Q. et al. Silencing TLR4/MyD88/NF-κB Signaling Pathway Alleviated Inflammation of Corneal Epithelial Cells Infected by ISE. Inflammation 44, 633–644 (2021). https://doi.org/10.1007/s10753-020-01363-1

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