Department of Stomatology, Zhumadian Central Hospital, 747 West Zhonghua Road, Zhumadian, 463000, Henan, China.
Naunyn Schmiedebergs Arch Pharmacol. 2024 Dec;397(12):10125-10141. doi: 10.1007/s00210-024-03270-w. Epub 2024 Jul 9.
Oral lichen planus (OLP) is a carcinogenic chronic inflammatory oral disease, which lacks effective treatments. Fraxin is an active ingredient of the traditional Chinese medicine Qin Pi, which has an anti-inflammatory effect, but its effect on OLP is unclear. The aim of this study was to investigate the therapeutic effect of fraxin on OLP and the underlying mechanism. Human immortalized keratinocytes (HaCat) were incubated with fraxin (10, 20, or 40 µM) for 48 h and then treated with 10 µg/mL LPS for 24 h. Cell viability and apoptosis were detected. Next, the interaction between OCT3 and FGF2 was predicted by online database and verified by Co-IP analysis. Fraxin, Ad-OCT3, sh-OCT3, and sh-FGF2 were, respectively, applied to treat LPS-incubated HaCat cells, and cell viability, apoptosis, and secretion of inflammatory factors were detected with MTT, flow cytometry, and ELISA assays. Then, the involvement of OCT3 and FGF2 in the prevention of fraxin on HaCat cells from LPS-induced cell apoptosis and inflammation was investigated through multiple rescue experiments. In addition, OLP models were constructed in VDR mice and NOD/SCID mice by injecting with human OLP pathological tissue homogenates to verify the therapeutic effect of fraxin on OLP. Fraxin treatment increased cell viability and reduced cell apoptosis and the secretion of IL-6 and TNF-α in a dose-dependent manner. OCT3 was significantly upregulated in oral mucosa tissues of OLP mice. OCT3 silencing inhibited LPS-induced cell apoptosis and secretion of inflammatory factors. Fraxin incubation reduced the expression of OCT3, and OCT3 interacted with FGF2 to upregulate FGF2 protein. FGF2 silencing reduced the expression of p-p65/NF-κB protein and improved LPS-induced cell apoptosis and secretion of inflammatory factors. OCT3 overexpression increased the expression of FGF2 and p-p65/NF-κB proteins, rh-FGF2 aggravated this effect, while FGF2-Neu-Ab reversed this effect. The results of in vivo experiments showed that fraxin alleviated cell apoptosis and inflammation in oral buccal mucosa tissues of OLP mice. Fraxin inhibited cell apoptosis and inflammation by suppressing OCT3-mediated activation of the FGF2/NF-κB pathway, alleviating the progression of OLP.
口腔扁平苔藓（OLP）是一种致癌的慢性炎症性口腔疾病，缺乏有效的治疗方法。秦皮素是中药秦皮的一种活性成分，具有抗炎作用，但对 OLP 的作用尚不清楚。本研究旨在探讨秦皮素对 OLP 的治疗作用及其机制。将人永生化角质形成细胞（HaCat）与秦皮素（10、20 或 40 μM）孵育 48 h，然后用 10 μg/mL LPS 处理 24 h。检测细胞活力和凋亡。接下来，通过在线数据库预测 OCT3 和 FGF2 之间的相互作用，并通过 Co-IP 分析进行验证。分别用秦皮素、Ad-OCT3、sh-OCT3 和 sh-FGF2 处理 LPS 孵育的 HaCat 细胞，用 MTT、流式细胞术和 ELISA 检测细胞活力、凋亡和炎症因子的分泌。然后，通过多种挽救实验研究 OCT3 和 FGF2 对秦皮素预防 LPS 诱导的 HaCat 细胞凋亡和炎症的作用。此外，通过向 VDR 小鼠和 NOD/SCID 小鼠注射人 OLP 病理组织匀浆构建 OLP 模型，验证秦皮素对 OLP 的治疗作用。秦皮素处理呈剂量依赖性地增加细胞活力，减少细胞凋亡和 IL-6 和 TNF-α 的分泌。OCT3 在 OLP 小鼠口腔黏膜组织中明显上调。OCT3 沉默抑制 LPS 诱导的细胞凋亡和炎症因子的分泌。秦皮素孵育降低了 OCT3 的表达，OCT3 与 FGF2 相互作用以上调 FGF2 蛋白。FGF2 沉默降低了 p-p65/NF-κB 蛋白的表达，并改善了 LPS 诱导的细胞凋亡和炎症因子的分泌。OCT3 过表达增加了 FGF2 和 p-p65/NF-κB 蛋白的表达，rh-FGF2 加重了这一作用，而 FGF2-Neu-Ab 则逆转了这一作用。体内实验结果表明，秦皮素减轻了 OLP 小鼠口腔颊黏膜组织中的细胞凋亡和炎症。秦皮素通过抑制 OCT3 介导的 FGF2/NF-κB 通路的激活来抑制细胞凋亡和炎症，从而缓解 OLP 的进展。
