MicroRNA Let-7i Regulates Innate TLR4 Pathways in Peripheral Blood Mononuclear Cells of Patients with Ankylosing Spondylitis

Liangjie Lu; Haiming Fang; Mengchao Gu; Huihan Wang; Qiuxia Yu; Aqiong Chen; Kai-Feng Gan

doi:10.2147/IJGM.S397160

MicroRNA Let-7i Regulates Innate TLR4 Pathways in Peripheral Blood Mononuclear Cells of Patients with Ankylosing Spondylitis

Int J Gen Med. 2023 Apr 19:16:1393-1401. doi: 10.2147/IJGM.S397160. eCollection 2023.

Authors

Liangjie Lu¹, Haiming Fang¹, Mengchao Gu¹, Huihan Wang², Qiuxia Yu³, Aqiong Chen³, Kai-Feng Gan¹

Affiliations

¹ Department of Orthopaedics, Li Huili Hospital Affiliated to Ningbo University, Ningbo, 315040, People's Republic of China.
² Department of Orthopaedics, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, 450007, People's Republic of China.
³ Department of Rheumatology, Li Huili Hospital Affiliated to Ningbo University, Ningbo, 315040, People's Republic of China.

Abstract

Purpose: This study aimed to compare the changes in the expression of microRNA Let-7i in peripheral blood mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS) and the correlation between Let-7i and innate pro-inflammatory factors. It is necessary to search for a new biomarker to guide the prognosis of AS.

Methods: A total of 10 patients with AS and 10 healthy volunteers were selected as AS and control groups, respectively. The expression levels of Let-7i, Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), and interferon-gamma (IFN-γ) in PBMCs were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB) to explore the relationship between Let-7i and pro-inflammatory factors. Furthermore, the relationship between Let-7i and TLR4 was determined by the luciferase reporter technology.

Results: The expression level of Let-7i in PBMCs of patients with AS was significantly lower than that of healthy control. The expression levels of TLR4, NF-κB, and IFN-γ in PBMCs derived from patients with AS were significantly higher than those of healthy control. The results show that Let-7i manipulation can regulate lipopolysaccharide (LPS)-induced TLR4 and IFN-γ expression in CD4+ T cells of patients with AS. The overexpression of Let-7i in T cells of patients with AS can suppress TLR4 and IFN-γ LPS-induced expression levels of cellular mRNA and protein. Let-7i can directly interfere TLR4-3'untranslated region (UTR) sequence and regulate the expression of the TLR4 gene in Jurkat T cells.

Conclusion: Let-7i may be involved in the pathogenesis of AS, and Let-7i expression in PBMCs may be helpful for the diagnosis and treatment of AS in the future.

Keywords: Let-7i; TLR4; ankylosing spondylitis; innate inflammatory.

Grants and funding

This study was supported by the Medical and Health Plan of Zhejiang(No.2023KY1025), “HuiLi Funded” Research Project(2022YB009) and Medical Science and Technology Project of Ningbo, China (No.2019Y04).