PKCβ expression contributes to M1 macrophage-induced impairment in the osteogenic differentiation of periodontal ligament stem cells.

IF 1.5 4区 生物学 Q4 CELL BIOLOGY
Yang Liu, Zhaocen Liu
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Abstract

Protein kinase Cβ (PRKCB) is expressed in THP-1 cells and has been found upregulated in periodontitis. Exploring the specific molecular mechanisms that promote the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) is beneficial to optimizing periodontal regeneration. THP-1 cells were induced to differentiate into macrophages, and the levels of PRKCB in macrophages with different phenotypes were examined, including PKC activity. The effect of pg-LPS induction on osteogenic differentiation of hPDLSCs was measured by measuring alkaline phosphatase, osteocalcin, osteogenic-related proteins, and mineralized nodules. Ruboxistaurin, an inhibitor of PRKCB, was used to treat M1 macrophages to examine its effect on macrophage polarization. Additionally, the cascade effect of ruboxistaurin on osteogenic differentiation was investigated by co-incubating hPDLSCs with medium from macrophages. The results indicated that PRKCB upregulation and increased PKC activity were induced in M1 macrophages upon stimulation with LPS/IFN-γ. pg-LPS resulted in decreased levels of osteogenic-related genes in hPDLSCs, accompanied by a decrease in mineralized nodules. PRKCB inhibitor reduced PKC activity, inhibited macrophage M1 polarization, and reduced M1-related inflammatory cytokine secretion. Exposure of hPDLSCs to M1 macrophage-derived conditioned medium impaired their osteogenic differentiation potentials, which was significantly attenuated by pretreatment of M1 macrophages with ruboxistaurin. Together, inhibition of PRKCB suppressed inflammatory M1 macrophage polarization, thus attenuating M1 macrophage-induced impairment in the osteogenic differentiation of hPDLSCs. These results provide a theoretical and scientific basis for optimizing the potential clinical application of hPDLSC therapy in periodontal regeneration.

PKCβ表达参与M1巨噬细胞诱导的牙周韧带干细胞成骨分化损伤。
蛋白激酶Cβ (PRKCB)在THP-1细胞中表达,并在牙周炎中被发现上调。探索促进人牙周韧带干细胞成骨分化的具体分子机制,有助于优化牙周再生。将THP-1细胞诱导分化为巨噬细胞,检测不同表型巨噬细胞中PRKCB水平,包括PKC活性。通过测定碱性磷酸酶、骨钙素、成骨相关蛋白和矿化结节来检测pg-LPS诱导hPDLSCs成骨分化的影响。采用PRKCB抑制剂Ruboxistaurin治疗M1巨噬细胞,观察其对巨噬细胞极化的影响。此外,通过将巨噬细胞培养液与hPDLSCs共孵育,研究了ruboxistaurin对成骨分化的级联效应。结果表明,LPS/IFN-γ刺激M1巨噬细胞可诱导PRKCB上调和PKC活性升高。pg-LPS导致hPDLSCs中成骨相关基因水平下降,并伴有矿化结节的减少。PRKCB抑制剂降低PKC活性,抑制巨噬细胞M1极化,减少M1相关炎性细胞因子分泌。将hPDLSCs暴露于M1巨噬细胞衍生的条件培养基中会损害其成骨分化潜能,而用鲁波西陶林预处理M1巨噬细胞可显著减弱其成骨分化潜能。总之,抑制PRKCB抑制炎性M1巨噬细胞极化,从而减弱M1巨噬细胞诱导的hPDLSCs成骨分化损伤。这些结果为优化hPDLSC治疗牙周再生的潜在临床应用提供了理论和科学依据。
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来源期刊
CiteScore
3.70
自引率
4.80%
发文量
96
审稿时长
3 months
期刊介绍: In Vitro Cellular & Developmental Biology - Animal is a journal of the Society for In Vitro Biology (SIVB). Original manuscripts reporting results of research in cellular, molecular, and developmental biology that employ or are relevant to organs, tissue, tumors, and cells in vitro will be considered for publication. Topics covered include: Biotechnology; Cell and Tissue Models; Cell Growth/Differentiation/Apoptosis; Cellular Pathology/Virology; Cytokines/Growth Factors/Adhesion Factors; Establishment of Cell Lines; Signal Transduction; Stem Cells; Toxicology/Chemical Carcinogenesis; Product Applications.
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