Tufail Muhammad, Zhang Jing, He Cai-Yun, Hu Jia-Ju, Liang Jie, Wan Wen-Dong, Huang Yu-Qi, Rao Zhen, Wu Hong, Jiang Can-Hua, Li Ning
Department of Oral and Maxillofacial Surgery, Center of Stomatology, Xiangya Hospital, Central South University, Changsha, China.
Department of Oral Medicine, Center of Stomatology, Xiangya Hospital, Central South University, Changsha, China; Institute of Oral Precancerous Lesions, Central South University, Changsha, China; Research Center of Oral and Maxillofacial Tumor, Xiangya Hospital, Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China.
Chem Biol Interact. 2025 Aug 30;421:111716. doi: 10.1016/j.cbi.2025.111716.
Oral submucous fibrosis (OSF) is a chronic fibrotic disease common in Asia-Pacific regions and strongly linked to long-term areca nut chewing. Fibroblast senescence is known to contribute to tissue fibrosis, but the molecular mechanisms behind it remain unclear. In this study, we explored how arecoline, a key component of areca nut, influences fibroblast behavior. Specifically, human oral fibroblasts were treated with 100 μM arecoline, which significantly increased the expression of the transcription factor PU.1(Purine-rich box 1) (∼3.5-fold mRNA, ∼3-fold protein). Notably, overexpression of PU.1 (encoded by SPI1) enhanced collagen production, while its inhibition reduced fibrotic markers. To further investigate, RNA sequencing and bioinformatics analysis identified BTG2 (B-cell Translocation Gene 2) as a downstream target of PU.1. Moreover, ChIP-qPCR confirmed PU.1 binding to the BTG2 promoter. Further assays, including western blotting, immunofluorescence, and SA-β-gal staining, showed that the PU.1-BTG2 axis promoted fibroblast senescence and collagen accumulation. Moreover, further analysis revealed that arecoline-treated fibroblasts produced more lactate, which led to increased lactylation of the transcription factor YY1(Yin Yang 1). Interestingly, YY1 was found to enhance PU.1-BTG2 transcription and drive fibrotic responses. Critically, mutation of YY1 at lysine 183 blocked its lactylation and eliminated its ability to activate PU.1 and BTG2. Finally, an arecoline-exposed OSF mouse model validated key in vivo aspects of this pathway, supporting its physiological relevance. Overall, we demonstrate that lactylated YY1 promotes fibroblast senescence and collagen deposition by activating the PU.1-BTG2 axis. This newly identified mechanism highlights potential therapeutic targets for treating OSF.
口腔黏膜下纤维化（OSF）是一种在亚太地区常见的慢性纤维化疾病，与长期咀嚼槟榔密切相关。已知成纤维细胞衰老会导致组织纤维化，但其背后的分子机制仍不清楚。在本研究中，我们探讨了槟榔的关键成分槟榔碱如何影响成纤维细胞的行为。具体而言，用100μM槟榔碱处理人口腔成纤维细胞，其显著增加了转录因子PU.1（富含嘌呤盒1）的表达（mRNA约3.5倍，蛋白质约3倍）。值得注意的是，PU.1（由SPI1编码）的过表达增强了胶原蛋白的产生，而其抑制则降低了纤维化标志物。为了进一步研究，RNA测序和生物信息学分析确定BTG2（B细胞易位基因2）为PU.1的下游靶点。此外，ChIP-qPCR证实PU.1与BTG2启动子结合。包括蛋白质印迹、免疫荧光和SA-β-半乳糖苷酶染色在内的进一步实验表明，PU.1-BTG2轴促进了成纤维细胞衰老和胶原蛋白积累。此外，进一步分析表明，经槟榔碱处理的成纤维细胞产生了更多的乳酸，这导致转录因子YY1（阴阳1）的乳酰化增加。有趣的是，发现YY1增强了PU.1-BTG2的转录并驱动纤维化反应。至关重要的是，YY1赖氨酸183位点的突变阻止了其乳酰化，并消除了其激活PU.1和BTG2的能力。最后，槟榔碱暴露的OSF小鼠模型验证了该通路在体内的关键方面，支持了其生理相关性。总体而言，我们证明乳酰化的YY1通过激活PU.1-BTG2轴促进成纤维细胞衰老和胶原蛋白沉积。这一新发现的机制突出了治疗OSF的潜在治疗靶点。
