恩格letin通过抑制NF-κB和MAPK信号通路减轻椎间盘退变中的炎症和细胞凋亡。

Engeletin Alleviates the Inflammation and Apoptosis in Intervertebral Disc Degeneration via Inhibiting the NF-κB and MAPK Pathways.

作者信息

Li Baixing, Yang Xiao, Zhang Pu, Guo Jiadong, Rong Kewei, Wang Xin, Cao Xiankun, Zhou Tangjun, Zhao Jie

机构信息

Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200011, People's Republic of China.

出版信息

J Inflamm Res. 2022 Oct 10;15:5767-5783. doi: 10.2147/JIR.S371809. eCollection 2022.

DOI:10.2147/JIR.S371809

PMID:36238766

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9553281/

Abstract

PURPOSE

Low back pain (LBP) induced by intervertebral disc degeneration (IDD) brings progressively painful status and impairs the normal daily living. Engeletin is a plant-derived medicine with anti-inflammation and antioxidant functions. Therefore, we aim to confirm its protective effects against the intervertebral disc degeneration in vivo and in vitro.

METHODS

The cytotoxicity of engeletin was validated by CCK-8 tests. Using the TNF-α to simulate the inflammation status in vitro, the expression of inflammatory mediators and MMP families were determined by qPCR, Western blotting and confocal microscopy. Cell apoptosis was analyzed by flow cytometry and TUNEL assay. The expression of apoptosis-related proteins was tested by Western blotting. The activation of NF-κB and MAPK pathways was evaluated by Western blotting and confocal microscopy. In vivo, percutaneous needle puncture was used to establish the IDD model in rat, and engeletin was administrated via intradiscal injection. The therapeutic effects of engeletin were detected through imaging and histology analysis.

RESULTS

Cell viability tests demonstrated there was little cytotoxicity of engeletin toward NP cells. Pretreatment with engeletin effectively ameliorate the TNF-α-induced up-regulation of inflammatory mediators and MMP families, promoting the anabolism of ECM meanwhile. Cell apoptosis was also attenuated with the addition of engeletin. We found that the activation of MAPK and NF-κB signaling pathways and the nuclear translocation of phosphorylated p65 and p38 were inhibited prominently with the treatment of engeletin which may be the potential molecular mechanism for its anti-inflammation effects. Finally, the IDD induced by percutaneous needle puncture was partially alleviated with the injection of engeletin in vivo.

CONCLUSION

As a natural compound with little cytotoxicity, engeletin possesses the outstanding anti-inflammation and anti-apoptosis effects in the process of IDD in vitro and in vivo, which may be a promising medicine for the prevention and treatment of IDD-related low back pain.

摘要

目的

椎间盘退变（IDD）引起的下腰痛（LBP）会导致疼痛状况逐渐加重，并损害正常日常生活。恩格letin是一种具有抗炎和抗氧化功能的植物源药物。因此，我们旨在确认其在体内和体外对椎间盘退变的保护作用。

方法

通过CCK-8试验验证恩格letin的细胞毒性。使用TNF-α在体外模拟炎症状态，通过qPCR、蛋白质免疫印迹法和共聚焦显微镜检测炎症介质和MMP家族的表达。通过流式细胞术和TUNEL测定法分析细胞凋亡。通过蛋白质免疫印迹法检测凋亡相关蛋白的表达。通过蛋白质免疫印迹法和共聚焦显微镜评估NF-κB和MAPK信号通路的激活情况。在体内，采用经皮针刺法建立大鼠IDD模型，并通过椎间盘内注射给予恩格letin。通过影像学和组织学分析检测恩格letin的治疗效果。

结果

细胞活力测试表明，恩格letin对髓核细胞的细胞毒性很小。恩格letin预处理可有效改善TNF-α诱导的炎症介质和MMP家族的上调，同时促进细胞外基质的合成代谢。添加恩格letin后细胞凋亡也得到减轻。我们发现，恩格letin治疗可显著抑制MAPK和NF-κB信号通路的激活以及磷酸化p65和p38的核转位，这可能是其抗炎作用的潜在分子机制。最后，体内注射恩格letin可部分缓解经皮针刺诱导的IDD。