Department of Emergency, Dali Bai Autonomous Prefecture People's Hospital, Dali 671000, China.
Department of Emergency, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, China.
Mediators Inflamm. 2024 Oct 10;2024:6618927. doi: 10.1155/2024/6618927. eCollection 2024.
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.
An 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-B65 (NF-B65), phospho-kappa B alpha(p-IB-), 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-1, IL-6, IL-18, and TNF- expression levels in AR42J cells were measured via ELISA method. The cell morphology was examined using the AO/EB method.
The experiment confirmed a significant increase in the activity of AR42J cells treated with various doses of baicalein. Moreover, IL-1, IL-6, TNF-, and IL-18 expression levels in AR42J cells were dramatically reduced ( < 0.05), while miR-224-5p level was obviously enhanced. The protein and gene expression of PARP1, NF-B65, p-IB-, IL-18R, GSDMD, ASC, NLRP3, and caspase-1 was obviously decreased ( < 0.05). Apoptosis in AR42J cells was significantly reduced with significant improvement in cell morphology.
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-B and NLPR3/ASC/caspase-1/GSDMD pathways.
黄芩素已被用于治疗与炎症相关的疾病；然而，其具体的作用机制尚不清楚。因此，我们研究了黄芩素对脂多糖诱导的 AR42J 胰腺腺泡细胞（PAC）损伤的保护作用，并确定了其保护作用的机制。
采用脂多糖（LPS）（1mg/L）诱导的 AR42J 细胞（AP）建立急性胰腺炎（AP）细胞模型，用 3-(4,5)-二甲基噻唑(-z-y1)-3,5-二苯基四氮唑溴盐（MTT）法测定相对存活率。流式细胞术评估 AR42J PAC 细胞的凋亡率。采用 WB 和 RT-PCR 法检测 miR-224-5p、多聚 ADP-核糖聚合酶-1（PARP1）、核转录因子-B65（NF-B65）、磷酸化-kappa B alpha（p-IB-）、白细胞介素（IL）-18R、NOD 样受体热蛋白域相关蛋白 3（NLRP3）、Gasdermin D（GSDMD）、凋亡相关斑点样蛋白含有 CARD（ASC）和半胱天冬酶-1 的 RNA 和蛋白表达。采用 ELISA 法测定 AR42J 细胞中白细胞介素（IL）-1、IL-6、IL-18 和肿瘤坏死因子（TNF-）的表达水平。采用 AO/EB 法观察细胞形态。
实验证实，用不同剂量黄芩素处理的 AR42J 细胞活性明显增加。此外，AR42J 细胞中白细胞介素（IL）-1、IL-6、TNF-和白细胞介素（IL）-18 的表达水平明显降低（ < 0.05），而 miR-224-5p 水平明显升高。PARP1、NF-B65、p-IB-、IL-18R、GSDMD、ASC、NLRP3 和半胱天冬酶-1 的蛋白和基因表达明显降低（ < 0.05）。AR42J 细胞凋亡明显减少，细胞形态明显改善。
黄芩素可能通过抑制炎症反应和细胞焦亡，显著减轻 LPS 诱导的 AR42J PAC 损伤。其作用方式可能与更高的 miR-224-5p 表达有关，该表达可抑制 PARP1/NF-B 和 NLPR3/ASC/caspase-1/GSDMD 通路。
