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miR-200a-3p 通过靶向 ZEB2 并激活 NF-κB 通路抑制人牙周膜干细胞成骨分化。

miR-200a-3p represses osteogenesis of human periodontal ligament stem cells by targeting ZEB2 and activating the NF-κB pathway.

机构信息

Department of Orthodontics, Stomatological Hospital, Southern Medical University, Guangzhou, China.

Department of Orthodontics, Qiannan Traditional Chinese Medical Hospital, School of Stomatology and Medicine, Qiannan Buyi and Miao Autonomous Prefecture, Duyun, China.

出版信息

Acta Odontol Scand. 2022 Mar;80(2):140-149. doi: 10.1080/00016357.2021.1964593. Epub 2021 Oct 9.

Abstract

OBJECTIVES

Human periodontal ligament stem cells (hPDLSCs) bear multilineage differentiation potential and represent the cytological basis of periodontal tissue regeneration. microRNA (miR) is accepted as a critical regulator of cell differentiation. This study explored the molecular mechanism of miR-200a-3p in osteogenesis of hPDLSCs.

MATERIAL AND METHODS

hPDLSCs were cultured and identified . miR-200a-3p expression during osteogenic differentiation of hPDLSCs was detected. hPDLSCs were transfected with miR-200a-3p mimic or miR-200a-3p inhibitor. Alkaline phosphatase (ALP) activity, calcified nodules and osteogenesis-related genes of hPDLSCs were measured. The binding relationship between miR-200a-3p and ZEB2 was predicted and verified. hPDLSCs were infected with sh-ZEB2, and then the osteogenic capacity was examined. miR-200a-3p inhibitor-transfected hPDLSCs were infected with sh-ZEB2. The key proteins of the NF-κB pathway were measured.

RESULTS

miR-200a-3p expression was downregulated during osteogenic differentiation of hPDLSCs. Upregulation of miR-200a-3p reduced ALP activity, calcified nodules and osteogenesis-related genes of hPDLSCs, while downregulation of miR-200a-3p facilitated the osteogenesis of hPDLSCs. miR-200a-3p targeted ZEB2. ZEB2 silencing repressed osteogenesis of hPDLSCs. ZEB2 silencing attenuated the promoting effect of miR-200a-3p inhibitor on osteogenesis of hPDLSCs. miR-200a-3p activated the NF-κB pathway by targeting ZEB2.

CONCLUSION

miR-200a-3p repressed osteogenesis of hPDLSCs by targeting ZEB2 and activating the NF-κB pathway. This study may offer insights for periodontal tissue regeneration engineering.

摘要

目的

人牙周膜干细胞(hPDLSCs)具有多向分化潜能,是牙周组织再生的细胞学基础。微小 RNA(miRNA)被认为是细胞分化的关键调节因子。本研究探讨了 miR-200a-3p 在 hPDLSCs 成骨分化中的分子机制。

材料与方法

培养并鉴定 hPDLSCs。检测 hPDLSCs 成骨分化过程中 miR-200a-3p 的表达。转染 miR-200a-3p 模拟物或 miR-200a-3p 抑制剂。检测 hPDLSCs 的碱性磷酸酶(ALP)活性、钙化结节和成骨相关基因。预测并验证 miR-200a-3p 与 ZEB2 的结合关系。感染 sh-ZEB2 后检测 hPDLSCs 的成骨能力。转染 miR-200a-3p 抑制剂的 hPDLSCs 感染 sh-ZEB2。检测 NF-κB 通路的关键蛋白。

结果

hPDLSCs 成骨分化过程中 miR-200a-3p 表达下调。上调 miR-200a-3p 降低 hPDLSCs 的 ALP 活性、钙化结节和成骨相关基因表达,而下调 miR-200a-3p 促进 hPDLSCs 的成骨。miR-200a-3p 靶向 ZEB2。沉默 ZEB2 抑制 hPDLSCs 的成骨。沉默 ZEB2 减弱了 miR-200a-3p 抑制剂对 hPDLSCs 成骨的促进作用。miR-200a-3p 通过靶向 ZEB2 激活 NF-κB 通路。

结论

miR-200a-3p 通过靶向 ZEB2 抑制 hPDLSCs 的成骨作用,并激活 NF-κB 通路。本研究可为牙周组织再生工程提供思路。

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