The involvement of DDX3X in compression-induced nucleus pulposus pyroptosis.
The involvement of DDX3X in compression-induced nucleus pulposus pyroptosis.
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DOI：
10.1016/j.bbrc.2023.02.074
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发表时间：
2023-03
影响因子：
3.1
通讯作者：
Shouyuan Chi;Suyun Li;Zhiqiang Xu;Guoyu Yang;Yu Song;Zhiwei Liao;Cao Yang;Xinghuo Wu
中科院分区：
文献类型：
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作者：
Shouyuan Chi;Suyun Li;Zhiqiang Xu;Guoyu Yang;Yu Song;Zhiwei Liao;Cao Yang;Xinghuo Wu
目的探讨DDX3X与髓核(NP)焦亡的关系。方法检测受压诱导的人髓核(NP)细胞和组织中的DDX3X和焦亡相关蛋白(Caspase-1、全长GSDMD、Cleaved GSDMD)。 DDX3X 通过基因转染过度表达或敲低。 Western blot法检测NLRP3、ASC及细胞焦亡相关蛋白的表达。采用ELISA法检测IL-1β和IL-18。采用HE染色和免疫组化法观察椎间盘退变大鼠模型中DDX3X、NLRP3、Caspase-1的表达。结果退变的NP组织中DDX3X、NLRP3、Caspase-1高表达。 DDX3X 的过度表达诱导 NP 细胞焦亡，并增加 NLRP3、IL-1β、IL-18 和焦亡相关蛋白的水平。 DDX3X 的敲低显示与 DDX3X 的过表达相反的趋势。 NLRP3抑制剂CY-09有效阻止IL-1β、IL-18、ASC、Pro-caspase-1、全长GSDMD和Cleaved GSDMD表达的上调。在压缩性椎间盘退变大鼠模型中观察到 DDX3X、NLRP3 和 Caspase-1 表达增加。结论我们的研究表明，DDX3X 通过上调 NLRP3 表达介导 NP 细胞焦亡，最终导致椎间盘退变（IDD）。这一发现加深了对IDD发病机制的理解，并为IDD提供了一个有前景的新型治疗靶点。
ObjectiveA study has been conducted to investigate the relationship between DDX3X and nucleus pulposus (NP) pyroptosis.MethodsDDX3X and pyroptosis-related proteins (Caspase-1, Full-length GSDMD, Cleaved GSDMD) were measured in compression-induced human NP cells and tissue. DDX3X was overexpressed or knocked down by gene transfection. The expressions of NLRP3, ASC, and pyroptosis-related proteins were detected by Western blot assay. IL-1β and IL-18 were detected by ELISA. HE staining and immunohistochemistry were used to observe the expression of DDX3X, NLRP3, and Caspase-1 in the rat model of compression-induced disc degeneration.ResultsDDX3X, NLRP3, and Caspase-1 were highly expressed in degenerated NP tissue. Overexpression of DDX3X induced pyroptosis in NP cells and increased levels of NLRP3, IL-1β, IL-18, and pyroptosis-related proteins. Knockdown of DDX3X showed an opposite trend to overexpression of DDX3X. The NLRP3 inhibitor CY-09 effectively prevented the up-regulation of the expression of IL-1β, IL-18, ASC, Pro-caspase-1, Full-length GSDMD, and Cleaved GSDMD. Increased expression of DDX3X, NLRP3, and Caspase-1 was observed in the rat model of compression-induced disc degeneration.ConclusionOur study showed that DDX3X mediates pyroptosis of NP cells by upregulating NLRP3 expression, which ultimately leads to intervertebral disc degeneration (IDD). This discovery deepens the understanding of IDD pathogenesis and provides a promising and novel therapeutic target for IDD.