甘草素减轻牙龈卟啉单胞菌脂多糖/三磷酸腺苷诱导的人牙龈成纤维细胞细胞焦亡，通过抑制 NF-κB/NLRP3/GSDMD 信号通路。

Isoliquiritigenin alleviates P. gingivalis-LPS/ATP-induced pyroptosis by inhibiting NF-κB/ NLRP3/GSDMD signals in human gingival fibroblasts.

机构信息

Department of Periodontology, The Affiliated Hospital of Qingdao University, Qingdao 266003, China; School of Stomatology, Qingdao University, Qingdao, Shandong, China.

Department of Periodontology, The Affiliated Hospital of Qingdao University, Qingdao 266003, China.

出版信息

Int Immunopharmacol. 2021 Dec;101(Pt B):108338. doi: 10.1016/j.intimp.2021.108338. Epub 2021 Nov 15.

DOI:10.1016/j.intimp.2021.108338

PMID:34794890

Abstract

OBJECTIVE

To investigate whether pyroptosis is induced by Porphyromonas gingivalis-lipopolysaccharide (P. gingivalis-LPS)/ adenosine triphosphate (ATP) through NF-κB/NLRP3/GSDMD signaling in human gingival fibroblasts (HGFs) and whether isoliquiritigenin (ISL) alleviates pyroptosis by inhibition of NF-κB/NLRP3/GSDMD signals.

DESIGN

Periodontitis was optimally simulated using a combination of P. gingivalis-LPS and ATP. The expression levels of genes and proteins of NF-κB, NLRP3 inflammasome, GSDMD, and IL-1β was characterized by qRT-PCR, western blotting and ELISA. The 2',7'‑dichlorodihydrofluorescein diacetate fluorescence probe was used to determine the intracellular ROS level. Hoechst 33342 and PI double staining, cytotoxicity assay, and caspase-1 activity assay were used to confirm the influence of ISL on pyroptosis in P. gingivalis-LPS/ATP-treated HGFs.

RESULTS

P. gingivalis-LPS/ATP stimulation significantly promoted expression of NF-κB, the NLRP3 inflammasome, GSDMD, and IL-1β at gene and protein levels. The proportion of membrane-damaged cells, caspase-1 activity, and the release of lactate dehydrogenase (LDH) were also elevated. However, pretreatment with ISL observably suppressed these effects.

CONCLUSIONS

P. gingivalis-LPS/ATP induced pyroptosis in HGFs by activating NF-κB/NLRP3/GSDMD signals and ISL attenuated P. gingivalis-LPS/ATP-induced pyroptosis by inhibiting these signals. This evidence may provide a new direction for the treatment of periodontitis.

摘要

目的

探讨牙龈卟啉单胞菌脂多糖（P. gingivalis-LPS）/三磷酸腺苷（ATP）是否通过 NF-κB/NLRP3/GSDMD 信号诱导人牙龈成纤维细胞（HGFs）发生细胞焦亡，以及异甘草素（ISL）是否通过抑制 NF-κB/NLRP3/GSDMD 信号来减轻细胞焦亡。

设计

采用牙龈卟啉单胞菌 LPS 和 ATP 联合最佳模拟牙周炎。通过 qRT-PCR、western blot 和 ELISA 检测 NF-κB、NLRP3 炎性小体、GSDMD 和 IL-1β 的基因和蛋白表达水平。采用 2'，7'-二氯二氢荧光素二乙酸荧光探针测定细胞内 ROS 水平。Hoechst 33342 和 PI 双重染色、细胞毒性测定和 caspase-1 活性测定用于确认 ISL 对 P. gingivalis-LPS/ATP 处理的 HGFs 中细胞焦亡的影响。