2. Academic Validation
  3. Liyan Kaiyin Formula relieves reflux pharyngitis by regulating M1 macrophage polarization via the NF-κB/NLRP3 pathway

Liyan Kaiyin Formula relieves reflux pharyngitis by regulating M1 macrophage polarization via the NF-κB/NLRP3 pathway

  • Immunobiology. 2025 Jul;230(4):153095. doi: 10.1016/j.imbio.2025.153095.
Lirong Wang 1,  Xuqing Chen 2,  Tianyu Xu 1,  Youpeng Fei 1,  Qi Yang 1,  Jingjuan An 1,  Zeqing Li 1,  Kunmin Wu 3
Affiliations

Affiliations

  • 1 Department of Otolaryngology, Jiangsu Provincial Second Chinese Medicine Hospital(The Second Affiliated Hospital of Nanjing University of Traditional Chinese Medicine), Nanjing 210017, Jiangsu Province, China.
  • 2 Department of Otolaryngology, Jiangsu Provincial Second Chinese Medicine Hospital(The Second Affiliated Hospital of Nanjing University of Traditional Chinese Medicine), Nanjing 210017, Jiangsu Province, China; Department of Pharmacy, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
  • 3 Department of Otolaryngology, Jiangsu Provincial Second Chinese Medicine Hospital(The Second Affiliated Hospital of Nanjing University of Traditional Chinese Medicine), Nanjing 210017, Jiangsu Province, China. Electronic address: wukmsjphtcm@ph-edu.cn.
PMID: 40602310 DOI: 10.1016/j.imbio.2025.153095
Abstract

Background: We aimed to investigate whether Liyan Kaiyin Formula (LYKYF) can relieve reflux pharyngitis in rats by regulating M1 macrophage polarization via the nuclear factor-κB (NF-κB)/NOD-like Receptor protein 3 (NLRP3) pathway.

Methods: Forty rats were randomized into a sham group, a laryngopharyngeal reflux disease (LPRD) group, a LYKYF group and a LYKYF+CHPG group (n = 10). Enzyme-linked immunosorbent assay was conducted to measure the serum levels of inflammatory factors interleukin-6 (IL-6), IL-1β and tumor necrosis factor-α (TNF-α). Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) were performed to measure the expressions of proteins implicated in NF-κB/NLRP3 pathway. Western blotting was also used for the detection of M1 macrophage markers (CD86 and iNOS).

Results: Compared to the sham group, TNF-α, IL-6 and IL-1β levels in the serum, proportion of M1 macrophages in pharyngeal tissues, p-NF-κB p65/p65 ratio, protein expressions of NLRP3, Caspase-1, Apoptosis-associated speck-like protein (ASC), cluster of differentiation 86 (CD86) and inducible nitric oxide synthase, and mRNA expressions of NF-κB p65, NLRP3, Caspase-1 and ASC in the LPRD group exhibited significant elevations (P < 0.05). Compared with the LYKYF group, the LYKYF+CHPG group had significant elevations in serum TNF-α, IL-6 and IL-1β levels, proportion of M1 macrophages in pharyngeal tissues, p-NF-κB p65/p65 ratio, protein expressions of NLRP3, Caspase-1, ASC, CD86 and iNOS, as well as NF-κB p65, NLRP3, Caspase-1 and ASC mRNA expressions (P < 0.05). The identified key target genes were significantly enriched in GO terms associated with signal transduction, protein phosphorylation regulation, and adaptive responses to external stimuli.

Conclusion: LYKYF may suppress M1 macrophage polarization through suppressing the NF-κB/NLRP3 pathway activation, thereby alleviating reflux pharyngitis in rats.

Keywords

Formula; Macrophage; Pathway; Polarization; Rat; Reflux pharyngitis.

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