Chicken Surfactant Protein A1 Activates Macrophages Phagocytosis and Attenuates LPS-induced Inflammatory Response Through the TLR4-mediated NF-кB Pathway

Overview

Journal Poult Sci

Publisher Elsevier

Date 2025 Jan 29

PMID 39879901

Authors

Qian Zhou

Haifeng Xiong

Hanwen Wu

Chenxiao Wang

Xinyuan Chen

Hongmei Liu

Affiliations

Soon will be listed here.

Abstract

Chicken surfactant protein A1 (cSP-A1) is a soluble C-type lectin found primarily in chicken lungs. Its function and other potential bioactivities are unclear. This study aimed to express, purify, and identify recombinant cSP-A1 (RcSP-A1), investigate its effects on chicken macrophage HD11 cells, and evaluate its ability to regulate the LPS-induced inflammatory response. The results showed that RcSP-A1 was produced in HEK 293F cells and could be purified using a Ni affinity column. The RcSP-A1 purified concentration was 7.5 µg/mL. Functional examinations showed that RcSP-A1 could aggregate all tested bacterial strains and led to a macrophage phagocytosis rate significantly higher than in the control (p < 0.01). Subsequently, HD11 cells, preincubated with various RcSP-A1 concentrations (12.5, 25, and 50 μg/mL) and 5 mM CaCl for 2 h, were stimulated by LPS (1 μg/mL) for 24 h. The results showed that RcSP-A1 significantly attenuated the stimulating effects of LPS on the transcription and protein expression levels of proinflammatory cytokines (IL-1β, IL-6, and TNF-α) and inhibited nitric oxide production. Mechanism studies demonstrated that RcSP-A1 exerted an anti-inflammatory effect on LPS-stimulated cells by down-regulating the expression of TLR4, MyD88, and p65, up-regulating the expression of IкB-α, and inhibiting the activation of the NF-кB signaling pathway. These findings suggested that RcSP-A1 promoted bacterial aggregation and phagocytosis and inhibited the LPS-induced inflammatory response in HD11 cells through the TLR4/NF-κB signaling pathway, displaying an important role in innate immune defense.

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