| 王宗帅,蔡显义,朱旭,等.Ghrelin通过调控NF-κB/NLRP3焦亡信号通路抑制前交叉韧带成纤维细胞焦亡的机制研究.骨科,2023,14(6): 553-558. |
| Ghrelin通过调控NF-κB/NLRP3焦亡信号通路抑制前交叉韧带成纤维细胞焦亡的机制研究 |
| Mechanism of Ghrelin Inhibiting Pyroptosis of Anterior Cruciate Ligament Fibroblasts by Regulating NF-κB/NLRP3 Pyroptosis Signaling Pathway |
| 投稿时间：2023-07-12 |
| DOI：10.3969/j.issn.1674-8573.2023.06.012 |
| 中文关键词: 胃饥饿素 细胞焦亡 前交叉韧带损伤 成纤维细胞 核因子κB NOD样受体热蛋白结构域相关蛋白3 |
| 英文关键词: Ghrelin Pyroptosis Anterior cruciate ligament injury Fibroblasts NF-κB NLRP3 |
| 基金项目:清远市社会发展领域自筹经费科技计划项目（202130） |
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| 中文摘要: |
| 目的 探究胃饥饿素（Ghrelin）调控核因子κB（nuclear factor kappa-B，NF-κB）/NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）信号通路在前交叉韧带成纤维细胞焦亡中的作用。方法 前交叉韧带成纤维细胞分为对照组、肿瘤坏死因子α（tumor necrotic factor-α，TNF-α）炎症模型组、TNF-α+Ghrelin干预组，通过CCK-8确定TNF-α和Ghrelin干预的剂量和时间，Transwell法检测细胞迁移能力，Western Blot检测细胞焦亡相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶-1（cysteinyl aspartate specific proteinase，Caspase-1）、白细胞介素18（Interleukin-18，IL-18）、白细胞介素1β（Interleukin-1β，IL-1β）、消皮素D（gasdermin D，GSDMD）的表达，q-PCR检测Caspase-1、IL-18、IL-1β、GSDMD mRNA表达水平，Western Blot检测磷酸化P65（p-P65）和NLRP3的表达。结果 绘制CCK-8结果曲线，确定Ghrelin的干预浓度为20 ng/mL，TNF-α干预浓度为20 ng/mL，干预时间都为48 h；与对照组相比，TNF-α炎症模型组的细胞迁移能力明显降低（P＜0.001），细胞焦亡相关蛋白表达显著增高（P＜0.05），p-P65和NLRP3的表达显著增高（P＜0.05），Ghrelin干预后细胞迁移能力明显提高（P＜0.001），细胞焦亡相关蛋白显著降低（P＜0.05），p-P65和NLRP3的表达显著降低（P＜0.05）。结论 Ghrelin能够显著抑制前交叉韧带成纤维细胞焦亡，改善其迁移能力，这可能是通过调控NF-κB/NLRP3实现的。 |
| 英文摘要: |
| Objective To explore the role of Ghrelin in regulating NF-κB/NLRP3 in pyroptosis of anterior cruciate ligament fibroblasts. Methods Anterior cruciate ligament fibroblasts were divided into control group, TNF-α inflammation model group, and TNF-α+Ghrelin intervention group. CCK-8 was used to determine the dose and time of TNF-α and Ghrelin intervention. Transwell method was used to detect cell migration ability, and Western blotting was used to detect cell pyroptosis, and the expression of related proteins (Caspase-1, IL-18, IL-1β, GSDMD). The q-PCR The mRNA expression of Caspase-1, IL-18, IL-1β and GSDMD was detected by q-PCR. Western blotting was used to detect the expression of p-P65 and NLRP3. Results The CCK-8 result curve revealed that the intervention concentration of Ghrelin and TNF-α was 20 ng/mL, and the intervention time was 48 h. Compared with the control group, the cell migration ability of the TNF-α inflammation model group was significantly decreased (P＜0.001), the expression of pyroptosis-related proteins was significantly increased (P＜0.05), and the expression of p-P65 and NLRP3 was significantly increased (P＜0.05). The cell migration ability was significantly improved after Ghrelin intervention (P＜0.001), pyroptosis-related proteins were significantly decreased (P＜0.05), and the expression of p-P65 and NLRP3 was significantly decreased (P＜0.05). Conclusion Ghrelin can significantly inhibit the pyroptosis of anterior cruciate ligament fibroblasts and improve their migration ability, which may be achieved by regulating NF-κB/NLRP3. |
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