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Int J Biol Sci 2022; 18(8):3178-3193. doi:10.7150/ijbs.69652 This issue Cite

Research Paper

Sublytic C5b-9 Induces CCL3/4 Production and Macrophage Accumulation in Thy-1N Rats via PKC-α/p65/IRF-8 Axis

Wenbo Wang^1#, Baomei Qian^1#, Chenhui Zhao², Mingyu Peng¹, Longfei Liu¹, Mengxiao Xie¹, Na Peng³, Qingling He¹, Shuai Ying¹, Yufeng Zhu⁴, Tao Wang⁵, Dajun Hu³, Dan Zhao¹, Jing Zhang¹, Yingwei Wang^1,6, Wen Qiu^1,6✉

1. Department of Immunology, and Key Laboratory of Immunological Environment and Disease, Nanjing Medical University, Nanjing, Jiangsu 211166, China
2. Department of Medicine, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China
3. Department of Rheumatology and Nephrology, The Second People's Hospital of China Three Gorges University, Yichang, China
4. Clinical Medical Science of the First Clinical Medical College, Nanjing Medical University, Nanjing, Jiangsu 211166, China
5. Department of Nephrology, The Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing, Jiangsu 211100, China
6. Key Laboratory of Antibody Technology of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China
^# These two authors contributed to this work equally.

✉ Corresponding author: Wen Qiu, PhD, Department of Immunology, and Key Laboratory of Immunological Environment and Disease, Nanjing Medical University, 101 Longmian Avenue, Nanjing, Jiangsu 211166, China. Tel.: +86-25-86869455 (O), E-mail: qiuwenedu.cn More

Abstract

Mesangioproliferative glomerulonephritis (MsPGN) is a common human kidney disease. Rat Thy-1 nephritis (Thy-1N) is an animal model widely used for the study of MsPGN. Thy-1N is not only sublytic C5b-9-dependent, but also related to pro-inflammatory cytokine production and macrophage (Mφ) accumulation in rat renal tissues. In this study, we found that the expression or phosphorylation of chemokine CCL3/4, CD68 (Mφ marker), IRF-8, PKC-α and NF-κB-p65 (p65) were all up-regulated both in the renal tissues of Thy-1N rats (in vivo) and in the glomerular mesangial cells (GMCs) upon sublytic C5b-9 stimulation (in vitro). Further experiments in vitro revealed that the phosphorylated PKC-α (p-PKC-α) could promote p65 phosphorylation, and then p-p65 enhanced IRF-8 expression through binding to IRF-8 promotor (-591 ~ -582 nt and -299 ~ -290 nt). Additionally, up-regulation or silencing of IRF-8 gene promoted or reduced CCL3/4 production, and then regulated Mφ chemotaxis. The underlying mechanism involved in IRF-8 binding to CCL3 promoter (-249 ~ -236 nt), which resulted in CCL3 gene transcription. The experiments in vivo showed that knockdown of renal PKC-α, p65, IRF-8 and CCL3/4 genes could inhibit CCL3/4 production, Mφ accumulation, GMC proliferation and proteinuria of Thy-1N rats. Furthermore, p-PKC-α, p-p65, IRF-8, CCL3/4 expression and Mφ accumulation were also increased in the renal tissues of MsPGN patients. Collectively, these findings indicate that sublytic C5b-9 induces CCL3/4 production and Mφ accumulation via PKC-α/p65/IRF-8 axis, and finally aggravates the pathological changes of MsPGN.

Keywords: mesangioproliferative glomerulonephritis, Thy-1 nephritis, sublytic C5b-9, IRF-8, CCL3/4, macrophage

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