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目的 探讨大黄素在脂多糖(LPS)诱导的人牙髓细胞(HDPCs)成牙本质向分化中的作用。方法 分离培养HDPCs，不同浓度的LPS、大黄素预处理HDPCs，Western blot检测细胞中成牙本质向分化相关基因ALP、RUNX2、DMP-1、DSPP的蛋白表达水平；生物信息学、小干扰RNA (si-RNA) 转染技术，分析并验证大黄素的下游细胞信号通路及作用靶点。结果 与LPS组相比，大黄素组ALP、RUNX2、DMP-1、DSPP的蛋白表达水平上升(P<0.05)；GO和KEGG富集分析显示，MAPK信号通路富集最显著；Western blot结果显示，与OIM+LPS组相比，OIM+LPS+Emodin组的p-ERK/ERK表达量上升(P<0.05)。生物信息学筛选出与大黄素在炎症环境下促进HDPCs成牙本质分化相关的前10个核心基因，包括CD44、ITGB2、SP1、TLR2、CCL2、CCL5、ITGAX、CD86、ICAM1、SYK。RT-qPCR结果显示，与CON组相比，OIM组CD44基因的mRNA表达量上升(P<0.01)。敲低CD44表达后，与si-NC组相比，si-CD44组ALP、RUNX2、DMP-1、DSPP的蛋白表达水平降低(均P<0.05)。结论 大黄素可显著促进LPS诱导的人牙髓细胞成牙本质向分化，其调控作用与上调CD44表达、激活ERK信号通路密切相关。
Abstract：Objective To investigate the effect of emodin on the odontoblastic differentiation of human dental pulp cells (HDPCs) induced by lipopolysaccharide (LPS). Methods HDPCs were isolated， cultured and pretreated with LPS and emodin at gradient concentrations. The protein expression levels of odontoblastic differentiation markers (ALP， RUNX2， DMP-1 and DSPP) were detected by Western blot. Bioinformatics analysis and small interfering RNA (siRNA) transfection were performed to identify and verify the downstream signaling pathways and targets of emodin. Results Compared with the LPS group， the protein expression levels of ALP， RUNX2， DMP-1 and DSPP significantly increased in the Emodin group (all P<0.05). GO and KEGG enrichment analysis showed that the MAPK signaling pathway was the most significantly enriched pathway. Western blot results demonstrated that the p-ERK/ERK expression ratio was elevated in the OIM+LPS+Emodin group compared with the OIM+LPS group (P<0.05). The top 10 hub genes associated with emodin-mediated promotion of HDPC odontoblastic differentiation under inflammatory conditions were screened via bioinformatics， including CD44， ITGB2， SP1， TLR2， CCL2， CCL5， ITGAX， CD86， ICAM1 and SYK. RT-qPCR results revealed that the mRNA expression of CD44 was upregulated in the OIM group compared to the CON group (P<0.01). After CD44 knockdown， the protein levels of ALP， RUNX2， DMP-1 and DSPP decreased in the si-CD44 group compared to the si-NC group (all P<0.05). Conclusion Emodin can significantly promote odontoblastic differentiation of LPS-challenged HDPCs， and its regulatory effect is closely related to the upregulation of CD44 expression and activation of the ERK signaling pathway.
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基本信息:
中图分类号:R781.3
引用信息:
[1]张晓蕾,李午丽,王元银.大黄素促牙髓细胞成牙本质向分化与CD44/ERK的相关性[J].安徽医科大学学报().
基金信息:
安徽省卫生健康科研项目(编号：AHWJ2022a012、AHWJ2023A20112)
2026-08-03
2026-08-03
2026-08-03