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目的 探究脂多糖(LPS)抑制成骨细胞的增殖及分化作用与调控核因子κB(NF-κB)信号通路之间的关系。方法 以MC3T3-E1细胞为研究对象，分别采用不同浓度LPS(0、200和600 ng·mL-1)及NF-κB特异性抑制剂BAY 11-7082干预细胞的增殖与分化过程。通过MTT法评估细胞增殖能力，结合碱性磷酸酶(ALP)活性染色及茜素红染色，量化细胞的成骨分化水平；应用荧光定量PCR(qPCR)和蛋白质免疫印迹技术，检测Ⅰ型胶原α1链(COL1A1)、骨钙素(OC)、NF-κB亚基p65及其磷酸化形式(p-p65)的基因转录与蛋白表达水平。结果 与对照组相比，LPS显著上调了MC3T3-E1细胞p65磷酸化水平，并以剂量依赖方式抑制了COL1A1和OC的基因转录与蛋白表达。同时，LPS处理组细胞的增殖能力、ALP活性及钙结节形成量均显著降低。此外，NF-κB特异性抑制剂BAY 11-7082能够有效缓解LPS诱导的成骨抑制作用，显著恢复细胞的成骨分化相关表型。结论 LPS可通过上调p65磷酸化水平，激活NF-κB信号通路，进而抑制成骨细胞的成骨分化。
Abstract：AIM To investigate the relationship between lipopolysaccharide(LPS) inhibition of osteoblast proliferation and differentiation and the regulation of nuclear factor-κB(NF-κB) signaling pathway. METHODS The proliferation and differentiation processes of MC3T3-E1 cells were intervened under the conditions of different concentrations of LPS(0, 200, and 600 ng·mL-1) or the presence of the NF-κB inhibitor BAY 11-7082. The osteogenic proliferation was detected using MTT assay, and the osteogenic differentiation was quantified using alkaline phosphatase(ALP) activity staining, and alizarin red staining of calcium nodules. Fluorescent quantitative PCR(qPCR) and Western Blotting were employed to detect the transcriptional and protein expression levels of COL1A1, osteocalcin(OC), p65, and phosphorylated p65(p-p65) in the cells. RESULTS Compared with the control group, LPS significantly up-regulated the phosphorylation level of p65 in MC3T3-E1 cells. Meanwhile, the transcription and expression of genes(COL1A1 and OC) and proteins(COL1A1 and OC) related to osteogenic differentiation were significantly inhibited. The levels of cell proliferation, ALP activity, and calcium nodule formation associated with the osteogenic differentiation phenotype were also significantly suppressed, and all these effects showed a dose-dependent relationship. Additionally, the NF-κB specific inhibitor BAY 11-7082 could effectively attenuate LPS-induced osteogenic inhibition and significantly restore the osteogenic differentiation-related phenotypes of the cells. CONCLUSION LPS inhibits the osteogenic differentiation of osteoblasts by activating the NF-κB signaling pathway through upregulating the phosphorylation level of p65.
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基本信息:
DOI：10.19577/j.1007-4406.2025.08.002
中图分类号:R965
引用信息:
[1]林忠,陈昊宇,谢群丽,等.脂多糖通过激活调控核因子κB信号通路抑制成骨细胞的增殖与分化[J].中国临床药学杂志,2025,34(08):569-575.DOI:10.19577/j.1007-4406.2025.08.002.
基金信息:
浙江省基础公益基金(编号LGD19H310001); 台州市科技计划(编号20ywa12); 上虞科技计划(编号2017-11)
2025-08-25
2025-08-25