Background: Intervertebral disc degeneration (IDD) is the most common diagnosis of patients with lower back pain. IDD is the underlying lesion of many spinal degenerative diseases; however, the role of cGAS/Sting/NLRP3 pathway and epigallocatechin gallate (EGCG) in the development of IDD remained unclear.
Methods: The expressions of cGAS, Sting and NLRP3 mRNA of intervertebral disc (IVD) samples from IDD patients and controls were detected by RT-PCR. The nucleus pulposus cells (NPCs) were induced by hydrogen peroxide (H₂O₂) and used as an in-vitro model. Both 5 μM and 25 μM EGCG treatment were used to detect the effect of EGCG on the in-vitro model. Cell viability was detected by the MTT method, and cell apoptosis and cell cycle would be detected by flow cytometry. Western blot was used in the detection of the expression of cGAS/Sting/NLRP3 as well as apoptosis-related protein level. ELISA was used in the detection of pro-inflammatory factors, including IL-1β, TNF-α, IL-6 and IL-10.
Results: The expressions of cGAS, Sting and NLRP3 mRNA were significantly increased in the IVD samples from IDD patients and NLRP3 was associated with cGAS and Sting. Advanced in-vitro study showed that H₂O₂ significantly increased the expression of cGAS, Sting and NLRP3 protein levels. Advanced experiments showed that EGCG treatment demonstrated significant protective effects in cell viability, apoptosis, cell cycle arrest and inflammatory status through down-regulation of cGAS/Sting/NLRP3 pathway.
Conclusion: It was shown that the cGAS, Sting and NLRP3 up-regulation was associated with the incidence of IDD. Our findings also suggest that EGCG treatment would provide anti-apoptosis, anti-inflammation and promote cell viability in H₂O₂ treatment-incubated NPCs through inhibiting cGAS/Sting/NLRP3 pathway.



中文翻译：

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Epigallocatechin-3-Gallate 通过抑制 cGAS/Sting/NLRP3 激活来保护 H2O2 诱导的髓核细胞凋亡和炎症。

背景：椎间盘退变（IDD）是腰痛患者最常见的诊断。IDD 是许多脊柱退行性疾病的潜在病变；然而，cGAS/Sting/NLRP3 通路和表没食子儿茶素没食子酸酯 (EGCG) 在 IDD 发展中的作用仍不清楚。
方法：采用RT-PCR检测IDD患者和对照组椎间​​盘（IVD）标本中cGAS、Sting和NLRP3 mRNA的表达。_{过氧化氢（H 2} O ₂）诱导髓核细胞（NPCs）) 并用作体外模型。5 μM 和 25 μM EGCG 处理均用于检测 EGCG 对体外模型的影响。MTT法检测细胞活力，流式细胞仪检测细胞凋亡和细胞周期。Western blot检测cGAS/Sting/NLRP3表达及凋亡相关蛋白水平。ELISA用于检测促炎因子，包括IL-1β、TNF-α、IL-6和IL-10。
结果： IDD患者体外诊断样本中cGAS、Sting和NLRP3 mRNA的表达显着增加，并且NLRP3与cGAS和Sting相关。先进的体外研究表明，H ₂ O ₂显着增加cGAS、Sting和NLRP3蛋白水平的表达。先进的实验表明，EGCG 治疗通过下调 cGAS/Sting/NLRP3 通路在细胞活力、细胞凋亡、细胞周期停滞和炎症状态方面表现出显着的保护作用。
结论：表明cGAS、Sting和NLRP3的上调与IDD的发生有关。我们的研究结果还表明，EGCG 治疗将通过抑制 cGAS/Sting/NLRP3 通路在 H ₂ O ₂治疗孵育的 NPC中提供抗细胞凋亡、抗炎和促进细胞活力。