Dysregulation of lncRNA TFAP2A-AS1 is involved in the pathogenesis of pulpitis by the regulation of microRNA-32-5p

IF 3.1 4区医学 Q3 IMMUNOLOGY Immunity, Inflammation and Disease Pub Date : 2024-09-10 DOI:10.1002/iid3.1312

Mingming Liu, Weijing Jia, Lin Bai, Qiaolin Lin

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Abstract

Objective

This study was designed to evaluate TFAP2A-AS1 expression in the dental pulp of teeth with or without pulpitis and to determine the function of TFAP2A-AS1 in pulp cells.

Methods

GSE92681 was analyzed to filter out differentially expressed lncRNAs. Pulp samples from teeth with pulpitis and healthy teeth (control) were examined using real-time (RT) quantitative polymerase chain reaction (qPCR). Human dental pulp stem cells (hDPSCs) were cultured in a specific medium for osteogenic induction, or treated with lipopolysaccharide (LPS) to simulate inflammation. The viability and apoptosis of human DPSCs (hDPSCs) were determined by XTT assay and apoptosis detection kit. Inflammation was induced by LPS and assessed by measuring the expression and release of inflammatory cytokines after TFAP2A-AS1 knockdown. Osteogenic differentiation of hDPSCs was investigated by determining expression levels of osteogenic markers and alkaline phosphatase (ALP) activity after TFAP2A-AS1 overexpression. The downstream microRNA (miRNA) was predicted. Dual-luciferase reporter was used to confirm the binding between miR-32-5p and TFAP2A-AS1.

Results

The expression of TFAP2A-AS1 was evaluated in inflamed pulp using RT-qPCR. TFAP2A-AS1 had a discriminatory ability for healthy individuals and patients with pulpitis. The expression of TFAP2A-AS1 decreased upon the osteogenic differentiation of hDPSCs, and increased upon the LPS induction. TFAP2A-AS1 can reverse the osteogenic differentiation of hDPSCs, as evidenced by decreased levels of dentine sialophosphoprotein, dentin matrix protein−1, and ALP activity. TFAP2A-AS1 knockdown can promote cell proliferation of hDPSCs and relieve LPS-induced inflammation, as evidenced by decreased levels of TNF-α, IL-1β, and IL-6. miR-32-5p was identified as a downstream miRNA of TFAP2A-AS1.

Conclusion

This study demonstrated the expression and potential function of TFAP2A-AS1 in the human dental pulp. TFAP2A-AS1 can inhibit odontogenic differentiation but promote inflammation in pulp cells.

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通过调控microRNA-32-5p，lncRNA TFAP2A-AS1的失调参与了牙髓炎的发病机制

目的本研究旨在评估 TFAP2A-AS1 在患有或未患有牙髓炎的牙髓中的表达情况，并确定 TFAP2A-AS1 在牙髓细胞中的功能。方法分析 GSE92681 以筛选出差异表达的 lncRNA。使用实时（RT）定量聚合酶链反应（qPCR）检测患有牙髓炎的牙齿和健康牙齿（对照组）的牙髓样本。在特定培养基中培养人牙髓干细胞（hDPSCs）以诱导成骨，或用脂多糖（LPS）处理以模拟炎症。人牙髓干细胞（hDPSCs）的存活率和凋亡率通过 XTT 试验和凋亡检测试剂盒进行测定。用 LPS 诱导炎症，并通过测量 TFAP2A-AS1 敲除后炎症细胞因子的表达和释放来评估炎症情况。通过测定TFAP2A-AS1过表达后成骨标志物的表达水平和碱性磷酸酶（ALP）活性，研究了hDPSCs的成骨分化。对下游微RNA（miRNA）进行了预测。使用双荧光素酶报告器确认 miR-32-5p 与 TFAP2A-AS1 之间的结合。结果使用 RT-qPCR 评估了发炎牙髓中 TFAP2A-AS1 的表达。TFAP2A-AS1 对健康人和牙髓炎患者具有鉴别能力。在 hDPSCs 成骨分化过程中，TFAP2A-AS1 的表达量减少，而在 LPS 诱导过程中，TFAP2A-AS1 的表达量增加。TFAP2A-AS1能逆转hDPSCs的成骨分化，这体现在牙本质矽磷蛋白、牙本质基质蛋白-1和ALP活性水平的降低上。敲除 TFAP2A-AS1 可促进 hDPSCs 的细胞增殖并缓解 LPS 诱导的炎症，这体现在 TNF-α、IL-1β 和 IL-6 水平的降低。结论本研究证明了 TFAP2A-AS1 在人类牙髓中的表达和潜在功能。TFAP2A-AS1 可抑制牙髓分化，但会促进牙髓细胞的炎症反应。

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来源期刊

Immunity, Inflammation and Disease Medicine-Immunology and Allergy

CiteScore

3.60

自引率

0.00%

发文量

146

审稿时长

8 weeks

期刊介绍： Immunity, Inflammation and Disease is a peer-reviewed, open access, interdisciplinary journal providing rapid publication of research across the broad field of immunology. Immunity, Inflammation and Disease gives rapid consideration to papers in all areas of clinical and basic research. The journal is indexed in Medline and the Science Citation Index Expanded (part of Web of Science), among others. It welcomes original work that enhances the understanding of immunology in areas including: • cellular and molecular immunology • clinical immunology • allergy • immunochemistry • immunogenetics • immune signalling • immune development • imaging • mathematical modelling • autoimmunity • transplantation immunology • cancer immunology