基于NLRP3/IL-18/NF-κB通路探讨化痰安神方改善神经元细胞凋亡的作用机制

doi:10.3969/j.issn.1006-7795.2025.03.013

首都医科大学学报 ›› 2025, Vol. 46 ›› Issue (3): 487-495.doi: 10.3969/j.issn.1006-7795.2025.03.013

• 精神疾病中西医结合治疗 • 上一篇下一篇

基于NLRP3/IL-18/NF-κB通路探讨化痰安神方改善神经元细胞凋亡的作用机制

李玥,陈俊逾^*,常春雷,张静

新疆维吾尔自治区中医医院神志病科，乌鲁木齐 830099

收稿日期:2025-03-04 出版日期:2025-06-21 发布日期:2025-06-25
通讯作者: 陈俊逾 E-mail:xjchenjunyu@163.com
基金资助:
新疆维吾尔自治区自然科学基金资助项目（2022D01C160）。

Investigation on the mechanism of Huatan Anshen Decoction to improve neuronal apoptosis based on NLRP3/IL-18/NF-κB pathway

Li Yue， Chen Junyu^*, Chang Chunlei, Zhang Jing

Department of Psychiatry, Hospital of Traditional Chinese Medicine of Xinjiang Uygur Autonomous Region, Urumqi 830099, China

Received:2025-03-04 Online:2025-06-21 Published:2025-06-25
Supported by:
This study was supported by Natural Science Foundation of Xinjiang Uygur Autonomous Region（2022D01C160）.

摘要/Abstract

摘要： 目的基于核因子激活的B细胞的NOD-样受体热蛋白结构域相关蛋白3（NOD-like receptor pyrin domain-containing protein 3, NLRP3）/白细胞介素－18（interleukin-18, IL-18）/κ－轻链（nuclear factor-activated B cell κ-light chain, NF-κB）信号通路探讨化痰安神汤抑制神经元细胞凋亡的作用机制。方法体外培养HT22小鼠海马神经元细胞，C57小鼠灌胃化痰安神方后制备含药小鼠血清，CCK-8法筛选化痰安神方含药血清最佳干预浓度。采用不同浓度含药血清处理HT22细胞后，予脂多糖（lipopolysaccharide,LPS）诱导其发生凋亡，CCK-8法L-乳酸脱氢酶（L-lactate dehydrogenase,L-LDH）生化检测法和Annexin V/PI 染色法用于检测细胞损伤与凋亡情况，Western blotting法用于测定通路相关蛋白P-NF-κB/NF-κB、凋亡相关的斑点样蛋白(apoptosis-associated speck-like protein containing, ASC)、NLRP3、Caspase-1的表达，酶联免疫吸附测定法用于测定IL-18的表达。过表达NLRP3后重复试验，进行单基因功能验证。结果化痰安神方含药血清可改善LPS诱导的细胞损伤和凋亡，降低细胞L-LDH浓度（P<0.05）；显著下调NLRP3/IL-18/NF-κB信号通路相关蛋白的转录表达（P<0.05）；显著降低细胞内IL-18浓度；同时过表达NLRP3可逆转化痰安神方含药血清对HT22细胞凋亡抑制作用（P<0.05）。结论化痰安神方能够通过抑制NLRP3/IL-18/NF-κB信号通路改善神经元细胞凋亡。

关键词: 化痰安神方, HT22细胞, 凋亡, NLRP3/IL-18/NF-κB信号通路, 炎症小体

Abstract: Objective To elucidate the mechanism by which Huatan Anshen decoction inhibits neuronal apoptosis through modulation of the NOD-like receptor pyrin domain-containing protein 3 (NLRP3)/interleukin-18 (IL-18) / nuclear factor-activated B cell κ-light chain (NF-κB) signaling pathway. Methods The HT22 cell line derived from mouse hippocampal neurons were cultured in vitro, and serum containing Huatan Anshen Decoction was prepared from C57 mice after intragastric administration. The optimal concentration of Huatan Anshen Decoction serum for intervention was determined with the CCK-8 method. Following treatment with different concentrations of drug-containing serum, lipopolysaccharide (LPS) was used to induce apoptosis in HT22 cells. Cell damage and apoptosis were assessed with the CCK-8 assay and Annexin V/PI staining. Western blotting and reverse transcription polymerase chain reaction was employed to measure the expression levels of NF-κB,apoptosis-associated speck-like protein containing (ASC), NLRP3, and Caspase-1, while enzyme-linked immunosorbent assay (ELISA) was used to detect IL-18 expression. The experiments was repeated after overexpression of NRLP3 to conduct single gene function verification. Results Serum containing Huatan Anshen Decoction significantly improved LPS-induced cellular damage and apoptosis in HT22 cells while reduced cellular L-lactate dehydrogenase, (L-LDH) levels (P<0.05).Transcriptional expression of proteins related to the NLRP3/IL-18/NF-κB signaling pathway showed significant downregulation(P<0.05). Additionally, it markedly decreased intracellular IL-18 levels. Furthermore, overexpression of NF-κB reversed the inhibitory effect of Huatan Anshen decoction-containing serum on HT22 cell apoptosis (P<0.05). Conclusion Huatan Anshen Decoction can improve neuronal apoptosis via suppressing the NLRP3/IL-18/NF-κB signaling pathway.

Key words: Huatan Anshen Decoction, HT22 cells, apoptosis, NLRP3/IL-18/NF-κB signaling pathway, inflammasome

中图分类号:

R741

李玥, 陈俊逾, 常春雷, 张静. 基于NLRP3/IL-18/NF-κB通路探讨化痰安神方改善神经元细胞凋亡的作用机制[J]. 首都医科大学学报, 2025, 46(3): 487-495.

Li Yue, Chen Junyu, Chang Chunlei, Zhang Jing. Investigation on the mechanism of Huatan Anshen Decoction to improve neuronal apoptosis based on NLRP3/IL-18/NF-κB pathway[J]. Journal of Capital Medical University, 2025, 46(3): 487-495.

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基于NLRP3/IL-18/NF-κB通路探讨化痰安神方改善神经元细胞凋亡的作用机制

Investigation on the mechanism of Huatan Anshen Decoction to improve neuronal apoptosis based on NLRP3/IL-18/NF-κB pathway

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