原儿茶酸在胶质瘤中的保护作用：通过NLRP3/半胱天冬酶-1/GSDMD轴调节细胞生长、迁移和焦亡

Protective Role of Protocatechuic Acid in Glioma: Modulation of Cell Growth, Migration, and Pyroptosis via NLRP3/Caspase-1/GSDMD Axis.

作者信息

Zhang Wen, Cai Yeyan, Zheng Haijun

机构信息

Department of Neurosurgery, The Third Affiliated Hospital of Wenzhou Medical University, 325200 Wenzhou, Zhejiang, China.

出版信息

Discov Med. 2025 Apr;37(195):659-668. doi: 10.24976/Discov.Med.202537195.57.

DOI:10.24976/Discov.Med.202537195.57

PMID:40287802

Abstract

BACKGROUND

Glioblastoma is a common primary malignant tumor posing a serious threat to human life and health. Protocatechuic acid (PCA) is a natural phenolic compound with good anti-tumor activity. The study aimed to investigate whether pyroptosis can be activated by PCA in glioma cell.

METHODS

Different concentrations of PCA were used to treat glioma cell lines U87 and U251 for varying durations. Cell proliferation was quantified using the Cell Counting Kit-8 (CCK-8) assay. The Transwell chamber assay was employed to evaluate cell invasion, while cell migration was assessed via the scratch assay. Pyroptosis levels were determined through immunofluorescence staining. Additionally, the protein and mRNA expression levels of nucleotide-binding and oligomerization domain-like receptor thermal protein domain-associated protein 3 (NLRP3), cysteinyl aspartate specific proteinase (caspase-1), and gasdermin D (GSDMD) were analyzed using Western blotting and quantitative reverse-transcription polymerase chain reaction (qRT-PCR).

RESULTS

Intervention with PCA resulted in a significant suppression of viability, invasion and migration of glioma cells in a dose-dependent manner ( < 0.05). Additionally, the GSDMD positivity rate, as well as the protein and mRNA expression levels of NLRP3, caspase-1, and GSDMD showed significant increases in glioma cells ( < 0.05). Further intervention with NLRP3-specific inhibitor MCC950 reversed the effects of PCA and resulted in a significant increase in cell viability and number of invading cells ( < 0.01), a significant decrease in GSDMD positivity ( < 0.01), and a significant decrease in the protein and mRNA expression levels of NLRP3, caspase-1, and GSDMD in glioma cells ( < 0.01).

CONCLUSION

PCA mediates pyroptosis in glioma cells by regulating the NLRP3/caspase-1/GSDMD signaling pathway.

摘要

背景

胶质母细胞瘤是一种常见的原发性恶性肿瘤，对人类生命健康构成严重威胁。原儿茶酸（PCA）是一种具有良好抗肿瘤活性的天然酚类化合物。本研究旨在探讨PCA是否能在胶质瘤细胞中激活细胞焦亡。

方法

用不同浓度的PCA处理胶质瘤细胞系U87和U251不同时间。使用细胞计数试剂盒-8（CCK-8）法对细胞增殖进行定量。采用Transwell小室实验评估细胞侵袭能力，通过划痕实验评估细胞迁移能力。通过免疫荧光染色测定细胞焦亡水平。此外，使用蛋白质免疫印迹法和定量逆转录聚合酶链反应（qRT-PCR）分析核苷酸结合寡聚化结构域样受体热蛋白结构域相关蛋白3（NLRP3）、半胱天冬酶-1（caspase-1）和gasdermin D（GSDMD）的蛋白质和mRNA表达水平。

结果

PCA干预导致胶质瘤细胞的活力、侵袭和迁移能力受到显著抑制，且呈剂量依赖性（<0.05）。此外，胶质瘤细胞中GSDMD阳性率以及NLRP3、caspase-1和GSDMD的蛋白质和mRNA表达水平均显著升高（<0.05）。用NLRP3特异性抑制剂MCC950进一步干预可逆转PCA的作用，导致细胞活力和侵袭细胞数量显著增加（<0.01），GSDMD阳性率显著降低（<0.01），胶质瘤细胞中NLRP3、caspase-1和GSDMD的蛋白质和mRNA表达水平显著降低（<0.01）。