时珍国医国药2025，Vol.36Issue(4)：628-635,8.DOI:10.70976/j.1008-0805.SZGYGY-2025-0405
大黄灵仙胶囊含药血清对胆管细胞炎症模型miRNA-140-5p表达及NF-κB/NLRP3信号通路的影响
Effect of drug-containing serum of Dahuang Lingxian Capsule on miRNA-140-5p expres-sion and NF-κB/NLRP3 signaling pathway in bile duct cell inflammation model
摘要
Abstract
Objective To explore the potential mechanisms of drug-containing serum of Dahuang Lingxian Capsule(DHLX)on miR-NA-140-5p expression and the NF-κB/NLRP3 signaling pathway in bile duct cells induced by lipopolysaccharide(LPS)in a rat model of inflammationMethods Twelve SD rats were randomly divided into the blank serum group and drug-containing serum group,with6 rats in each group.The drug-containing serum group was given320 mg·kg-1·d-1 intragastric administration,while the blank serum group received an equal volume of distilled water.Treatment was given once daily for 3 consecutive days,and the drug-contai-ning serum was prepared by taking blood from abdominal aorta 6 hours after the final administration.Bile duct cells were divided into the blank group,model group,DHLX group,DHLX+inhibitor group and DHLX+inhibitor control group.Cells from each group were col-lected,and the morphological changes in bile duct cells were observed under electron microscope.Cell Caspase-1 activity was detected in each group.Levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)were determined by ELISA.qRT-PCR was used to determine the expression levels of miRNA-140-5p,tumor necrosis factor α(TNF-α),interleukin-6(IL-6),Toll-like receptor 4(TLR4),nuclear factor κB p65(NF-κB p65),NOD-like receptor pyrin domain-containing protein 3(NLRP3),Caspase-1,and apoptosis-associated speck-like protein(ASC)mRNA.Western blot(WB)was performed to detect the protein expression levels of TNF-α,IL-6,TLR4,NF-κB p65,NLRP3,Caspase-1,and ASC.Results Compared with blank group,the model group exhib-ited nuclear contraction,cell membrane vesiculation,cytoplasmic vacuolation,blurred or reduced mitochondrial cristae,endoplasmic re-ticulum expansion,and Caspase-1 activity was significantly increased(P<0.01).The mRNA and protein expressions of IL-1β,IL-18,TNF-α,IL-6,TLR4,NF-κB p65,NLRP3,Caspase-1 and ASC were increased(P<0.05),while the expression of miR-NA-140-5p was significantly decreased(P<0.01).Compared with the model group,bile duct cell injury was reduced in the DHLX group,with a significant decrease in Caspase-1 activity(P<0.01).The mRNA and protein expressions of IL-1β,IL-18,TNF-α,IL-6,TLR4,NF-κB p65,NLRP3,Caspase-1 and ASC were decreased(P<0.05),while the expression of miRNA-140-5p was significantly increased(P<0.01).Compared with the DHLX group,bile duct cell injury was exacerbated in the DHLX+inhibitor group,with a significant increase in Caspase-1 activity(P<0.01).The mRNA and protein expressions of IL-1β,IL-18,TNF-α,IL-6,TLR4,NF-κB p65,NLRP3,Caspase-1 and ASC were increased(P<0.05),while the expression of miRNA-140-5p was significantly decreased(P<0.01).There were no statistically significant differences in the above parameters between the DHLX+in-hibitor control group and the DHLX+inhibitor group(P>0.05).Compared with the DHLX+inhibitor group,bile duct cell pyropto-sis was significantly relieved in the DHLX+inhibitor control group,with a notable decrease in Caspase-1 activity(P<0.01).The mRNA and protein levels of IL-1β,IL-18,TNF-α,IL-6,TLR4,NF-κB p65,NLRP3,Caspase-1,and ASC were reduced(P<0.05),while miRNA-140-5p expression was significantly increased(P<0.01).Conclusion DHLX may play a role in the pre-vention and treatment of cholelithiasis by up-regulating the expression of miRNA-140-5p in bile duct cells,inhibiting the release of factors related to the NF-κB/NLRP3 signaling pathway,suppressing pyroptosis,and delaying the progression of cellular inflammation.关键词
胆石症/胆管细胞/大黄灵仙胶囊/miRNA-140-5p/NF-κB/NLRP3信号通路Key words
Cholelithiasis/Bile duct cells/Dahuang Lingxian Capsule/miRNA-140-5p/NF-κB/NLRP3 signaling pathway分类
医药卫生引用本文复制引用
居燕飞,陈雅璐,林龙,劳永彩,王清坚..大黄灵仙胶囊含药血清对胆管细胞炎症模型miRNA-140-5p表达及NF-κB/NLRP3信号通路的影响[J].时珍国医国药,2025,36(4):628-635,8.基金项目
国家自然科学基金(82060867 （）
82474507) （）
广西自然科学基金(2024GXNSFDA010025 （）
2024GXNSFBA010106) （）
广西重点研发计划项目(桂科AB24010130) （桂科AB24010130）
广西名中医传承工作室建设项目(GZY2024011) （GZY2024011）
广西壮族自治区研究生教育创新计划项目(YCSW2022350 （）
YCSY2023037) （）
