黄芩苷通过下调 miR-224-5p/PARP1 减轻炎症和脓毒症对脂多糖诱导的 AR42J PACs 的保护作用

IF 4.4 3区 医学 Q2 CELL BIOLOGY Mediators of Inflammation Pub Date : 2024-10-10 eCollection Date: 2024-01-01 DOI:10.1155/2024/6618927
Ming-Wei Liu, Chun-Hai Zhang, Shou-Hong Ma, De-Qiong Zhang, Li-Qiong Jiang, Yang Tan
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引用次数: 0

摘要

背景:黄芩素一直被用于治疗炎症相关疾病,但其具体作用机制尚不清楚。因此,我们研究了黄芩苷对脂多糖诱导的 AR42J 胰腺尖突细胞(PACs)损伤的保护作用,并确定了其保护作用的机制:方法:利用脂多糖(LPS)(1 mg/L)诱导的急性胰腺炎(AP)体外细胞模型(AR42J),采用3-(4,5)-二甲基噻唑(-z-y1)-3,5-二苯基四氮唑(MTT)技术测定相对存活率。流式细胞术用于评估 AR42J PACs 的凋亡率。miR-224-5p、聚ADP-核糖聚合酶-1(PARP1)、核转录因子-κB65(NF-κB65)、phospho-kappa B alpha(p-IκB-α)、白细胞介素(IL)-18R、NOD样受体热蛋白结构域-α(p-IκB-α)的RNA和蛋白表达量均有显著变化、根据 WB 和 RT-PCR 检测,检测到了 NOD 样受体热蛋白结构域相关蛋白 3(NLRP3)、gasdermin D(GSDMD)、含 CARD 的凋亡相关斑点样蛋白(ASC)和 caspase-1。通过 ELISA 方法检测了 AR42J 细胞中 IL-1β、IL-6、IL-18 和 TNF-α 的表达水平。采用 AO/EB 法检测细胞形态:结果:实验证实,用不同剂量的黄芩苷处理 AR42J 细胞后,其活性明显提高。此外,AR42J细胞中IL-1β、IL-6、TNF-α和IL-18的表达水平显著降低(P < 0.05),而miR-224-5p的表达水平明显提高。PARP1、NF-κB65、p-IκB-α、IL-18R、GSDMD、ASC、NLRP3和caspase-1的蛋白和基因表达明显降低(P < 0.05)。AR42J细胞凋亡明显减少,细胞形态明显改善:结论:黄芩苷可通过抑制炎症反应和热凋亡,明显减轻 LPS 诱导的 AR42J PAC 损伤。其作用模式可能与 miR-224-5p 的高表达有关,miR-224-5p 可抑制 PARP1/NF-κB 和 NLPR3/ASC/caspase-1/GSDMD 通路。
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Protective Effects of Baicalein on Lipopolysaccharide-Induced AR42J PACs through Attenuation of Both Inflammation and Pyroptosis via Downregulation of miR-224-5p/PARP1.

Background: Baicalein has been used to treat inflammation-related diseases; nevertheless, its specific mechanism of action is unclear. Therefore, we examined the protective effects of baicalein on lipopolysaccharide-induced damage to AR42J pancreatic acinar cells (PACs) and determined its mechanism of action for protection.

Methods: An in vitro cell model of acute pancreatitis (AP) was established using lipopolysaccharide (LPS) (1 mg/L)-induced PACs (AR42J), and the relative survival rate was determined using the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) technique. Flow cytometry was applied to evaluate the apoptotic rates of AR42J PACs. The RNA and protein expression of miR-224-5p, poly ADP-ribose polymerase-1 (PARP1), nuclear transcription factor-κB65 (NF-κB65), phospho-kappa B alpha(p-IκB-α), interleukin(IL)-18R, NOD-like receptor thermal protein domain-associated protein 3 (NLRP3), gasdermin D (GSDMD), apoptosis-associated speck-like protein containing a CARD (ASC), and caspase-1 was detected based on the WB and RT-PCR assays. IL-1β, IL-6, IL-18, and TNF-α expression levels in AR42J cells were measured via ELISA method. The cell morphology was examined using the AO/EB method.

Results: The experiment confirmed a significant increase in the activity of AR42J cells treated with various doses of baicalein. Moreover, IL-1β, IL-6, TNF-α, and IL-18 expression levels in AR42J cells were dramatically reduced (P  < 0.05), while miR-224-5p level was obviously enhanced. The protein and gene expression of PARP1, NF-κB65, p-IκB-α, IL-18R, GSDMD, ASC, NLRP3, and caspase-1 was obviously decreased (P < 0.05). Apoptosis in AR42J cells was significantly reduced with significant improvement in cell morphology.

Conclusion: Baicalein may significantly alleviate LPS-induced AR42J PAC damage by inhibiting the inflammatory response and pyroptosis. Its mode of action might be linked to higher miR-224-5p expression, which inhibits the PARP1/NF-κB and NLPR3/ASC/caspase-1/GSDMD pathways.

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来源期刊
Mediators of Inflammation
Mediators of Inflammation 医学-免疫学
CiteScore
8.70
自引率
0.00%
发文量
202
审稿时长
4 months
期刊介绍: Mediators of Inflammation is a peer-reviewed, Open Access journal that publishes original research and review articles on all types of inflammatory mediators, including cytokines, histamine, bradykinin, prostaglandins, leukotrienes, PAF, biological response modifiers and the family of cell adhesion-promoting molecules.
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