| 贺 莹,杨一帆,褚晓月,郭 静,王家亮.血根碱通过NF-κB信号通路调控牙周膜干细胞成骨分化[J].,2024,(11):2020-2026 |
| 血根碱通过NF-κB信号通路调控牙周膜干细胞成骨分化 |
| Sanguinarine Regulates Osteogenic Differentiation of Periodontal Ligament Stem Cells through NF-κB Signal Pathway |
| 投稿时间：2023-11-28 修订日期：2023-12-23 |
| DOI：10.13241/j.cnki.pmb.2024.11.004 |
| 中文关键词: 血根碱 炎性微环境 牙周膜干细胞 肿瘤坏死因子-α 核因子-κB 成骨分化 |
| 英文关键词: Sanguinarine Inflammatory microenvironment Periodontal ligament stem cells Tumor necrosis factor-α Nuclear factor-κB Osteogenic differentiation |
| 基金项目:陕西省重点研发项目计划（2018SF-111）；陕西省卫生健康科研基金项目（2022D050） |
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| 摘要 目的：探究血根碱（SAN）对肿瘤坏死因子-α（TNF-α）处理的人牙周膜干细胞（hPDLSCs）成骨分化的影响及机制。方法：将hPDLSCs分为6组：Control组、TNF-α组、0.1SAN组、1SAN组、10SAN组和100SAN组，所有hPDLSCs均用成骨诱导培养液培养。除Control组外，其他组细胞培养液中均添加10 ng/mL的TNF-α。0.1SAN组、1SAN组、10SAN组和100SAN组细胞培养液中分别添加0、0.1、1、10和100 μmol/L的血根碱。各组hPDLSCs均在37℃、5% CO2条件下培养21 d。通过可见光比色法检测碱性磷酸酶（ALP）活性。通过茜素红染色观察钙化结节形成，并统计OD562 nm （代表钙化结节形成量）。通过qRT-PCR检测Runt相关转录因子2（RUNX2）、骨钙素（OCN）、osterix（OSX）、牙骨质附着蛋白（CAP）、Smad4转录水平。通过Western blot检测核因子-κB（NF-κB）p65磷酸化水平。结果：与Control组比较，TNF-α组细胞的相对ALP活性降低和钙化结节形成量以及RUNX2、OCN、OSX、CAP和Smad4的mRNA相对表达量降低（P<0.05），p-NF-κB p65/NF-κB p65升高（P<0.05）。与TNF-α组比较，1SAN组、10SAN组和100SAN组的相对ALP活性和钙化结节形成量以及RUNX2、OCN、OSX、CAP和Smad4的mRNA相对表达量升高（P<0.05），p-NF-κB p65/NF-κB p65降低（P<0.05）。结论：血根碱可促进TNF-α处理的hPDLSCs的成骨分化，其机制可能与抑制NF-κB的激活有关，血根碱可能是促进炎性微环境中hPDLSCs成骨分化的候选药物。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect and mechanism of sanguinarine (SAN) on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) treated with tumor necrosis factor-α (TNF-α). Methods: hPDLSCs were divided into 6 groups: Control group, TNF-α group, TNF-α+0.1SAN group, TNF-α+1SAN group, TNF-α+10SAN group and TNF-α+100SAN group. All hPDLSCs were cultured in osteogenic induction medium. Except Control group, 10 ng/mL TNF-α was added to the culture medium of other groups. 0, 0.1, 1, 10, 100 μmol/L sanguinarine were added to the culture medium of TNF-α+0.1SAN group, TNF-α+1SAN group, TNF-α+10SAN group and TNF-α+100SAN group, respectively. HPDLSCs of all groups were cultured at 37 ℃ and 5% CO2 for 21 days. The activity of alkaline phosphatase (ALP) was detected by visible light colorimetry. The formation of calcified nodules was observed by alizarin red staining, and OD562 nm (representing the amount of calcified nodules) was counted. The transcription levels of Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), osterix (OSX), cementum attachment protein (CAP) and Smad4 were detected by qRT-PCR. The phosphorylation level of NF-kappa B (NF-κB) p65 was detected by Western blot. Results: Compared with that in the Control group, the relative ALP activity, amount of calcified nodules, and the relative expression of RUNX2, OCN, OSX, CAP and Smad4 mRNA in TNF-α group decreased (P<0.05), while p-NF-κB p65/NF-κB p65 increased (P<0.05). Compared with that in the TNF-α group, the relative ALP activity, amount of calcified nodules, RUNX2, OCN, OSX, CAP and Smad4 mRNA expression of TNF-α+1SAN group, TNF-α+10SAN group and TNF-α+100SAN group increased (P<0.05), while p-NF-κB p65/NF-κB p65 decreased (P<0.05). Conclusion: Sanguinarine can promote the osteogenic differentiation of hPDLSCs treated with TNF-α, and the mechanism may be related to the inhibition of the activation of NF-κB. Sanguinarine may be a candidate drug to promote the osteogenic differentiation of hPDLSCs in inflammatory microenvironment. |
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