Wang Lirong, Chen Xuqing, Xu Tianyu, Fei Youpeng, Yang Qi, An Jingjuan, Li Zeqing, Wu Kunmin
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 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.
Immunobiology. 2025 Jun 28;230(4):153095. doi: 10.1016/j.imbio.2025.153095.
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.
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).
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.
LYKYF may suppress M1 macrophage polarization through suppressing the NF-κB/NLRP3 pathway activation, thereby alleviating reflux pharyngitis in rats.
我们旨在研究利咽开音方（LYKYF）是否能通过核因子-κB（NF-κB）/Nod样受体蛋白3（NLRP3）途径调节M1巨噬细胞极化，从而缓解大鼠反流性咽炎。
将40只大鼠随机分为假手术组、喉咽反流病（LPRD）组、LYKYF组和LYKYF+CHPG组（n = 10）。采用酶联免疫吸附测定法检测血清中炎症因子白细胞介素-6（IL-6）、IL-1β和肿瘤坏死因子-α（TNF-α）的水平。采用蛋白质印迹法和逆转录-聚合酶链反应（RT-PCR）检测NF-κB/NLRP3途径相关蛋白的表达。蛋白质印迹法还用于检测M1巨噬细胞标志物（CD86和诱导型一氧化氮合酶）。
与假手术组相比，LPRD组血清中TNF-α、IL-6和IL-1β水平、咽组织中M1巨噬细胞比例、p-NF-κB p65/p65比值、NLRP3、半胱天冬酶-1、凋亡相关斑点样蛋白（ASC）、分化簇86（CD86）和诱导型一氧化氮合酶的蛋白表达以及NF-κB p65、NLRP3、半胱天冬酶-1和ASC的mRNA表达均显著升高（P < 0.05）。与LYKYF组相比，LYKYF+CHPG组血清中TNF-α、IL-6和IL-1β水平、咽组织中M1巨噬细胞比例、p-NF-κB p65/p65比值、NLRP3、半胱天冬酶-1、ASC、CD86和诱导型一氧化氮合酶的蛋白表达以及NF-κB p65、NLRP3、半胱天冬酶-1和ASC的mRNA表达均显著升高（P < 0.05）。鉴定出的关键靶基因在与信号转导、蛋白质磷酸化调节和对外界刺激的适应性反应相关的基因本体（GO）术语中显著富集。
LYKYF可能通过抑制NF-κB/NLRP3途径激活来抑制M1巨噬细胞极化，从而缓解大鼠反流性咽炎。
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