摘要
目的 基于Tol样受体4(Tol-like receptor 4,TLR4)/核因子-κB(nuclear factor-κB,NF-κB)信号通路探讨百合皂苷对脂多糖(lipopolysaccharide,LPS)诱导的人支气管肺上皮细胞(human bronchopulmonary epithelial cells,BEAS-2B)的影响。方法 于2024年10月—2025年2月在吉林省人民医院精准分子医学中心通过培养、传代正常BEAS-2B细胞,以及通过LPS构建肺部炎症损伤模型,采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)方法测定细胞存活率,实时荧光定量逆转录聚合酶链式反应(real-time quantitative reverse transcription polymerase chain reaction,RT-qPCR)检测NF-κB等基因m RNA的表达,Western blotting法检测NF-κB等相关蛋白表达。结果 CCK8法检测结果显示,不同浓度的LPS处理BEAS-2B细胞后,随着LPS浓度的增加,存活细胞的数量逐渐减少,当LPS浓度为0、25%、40%、55%、70%、85%时,吸光度(optical density,OD)值分别为(0.88±0.02)(0.87±0.01)(0.73±0.01)(0.51±0.03)(0.42±0.01)(0.25±0.02)。而在不改变LPS浓度的情况下用百合皂苷处理后结果显示存活细胞的数量较前增多,OD值分别升高至(0.90±0.02)(0.88±0.03)(0.79±0.02)(0.67±0.02)(0.48±0.04)(0.40±0.02)。RT-qPCR法检测结果显示,实验组1(百合皂苷浓度60%)NF-κB基因表达量为(0.46±0.03),低于对照组的(1.00±0.05),差异有统计学意义(P<0.05),实验组2(百合皂苷浓度90%)NF-κB基因表达量为(0.72±0.04),低于对照组的(1.00±0.05),差异有统计学意义(P<0.001)。Western blotting法检测结果显示,实验组1和实验组2目标蛋白NF-κB表达水平随着百合皂苷增高而降低。结论 百合皂苷可能通过调控TLR4/NF-κB信号通路改善LPS诱导的BEAS-2B细胞的损伤。
Objective To explore the effect of lilium saponins on lipopolysaccharide(LPS)-induced human bronchopulmonary epithelial cells(BEAS-2B)based on the Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.Methods From October 2024 to February 2025,at the precision molecular medicine center of Jilin Province People′s Hospital,normal BEAS-2B cells were cultured and passaged.An LPS-induced pulmonary inflammatory injury model was established.Cell viability was measured using the cell counting kit-8(CCK-8)assay.The mRNA expression levels of NF-κB and related genes were analyzed by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR),and protein expression levels of NF-κB were detected by Western blotting.Results The CCK8 assay results showed that after treating BEAS-2B cells with different concentrations of LPS,the number of surviving cells gradually decreased with the increase of LPS concentration.At LPS concentrations of 0,25%,40%,55%,70%,and 85%,the optical density(OD)values were(0.88±0.02)(0.87±0.01)(0.73±0.01)(0.51±0.03)(0.42±0.01)and(0.25±0.02),respectively.Subsequent treatment with lilium saponin without altering LPS concentrations resulted in increased cell viability,with elevated OD values of(0.90±0.02)(0.88±0.03)(0.79±0.02)(0.67±0.02)(0.48±0.04)and(0.40±0.02)correspondingly.RT-qPCR analysis demonstrated that the NF-κB gene expression level in experimental group 1(60%saponin concentration)was(0.46±0.03),which was lower than(1.00±0.05)in the control group,the difference was statistically significant(P<0.05).The expression level of NF-κB gene in experimental group 2(90%saponin concentration)was(0.72±0.04),which was lower than(1.00±0.05)in the control group,and the difference was statistically significant(P<0.001).The results of Western blotting detection showed that the expression levels of the target protein NF-κB in experimental group 1 and experimental group 2 decreased significantly with the increase of lilium saponins.Conclusion Lilium saponins may ameliorate LPS-induced BEAS-2B cell injury by regulating the TLR4/NF-κB signaling pathway.
作者
包寅秋
王丽丽
李丹
韩成勋
王平平
李玲
芦小单
张维杰
BAO Yinqiu;WANG Lili;LI Dan;HAN Chengxun;WANG Pingping;LI Ling;LU Xiaodan;ZHANG Weijie(School of Clinical Medicine,Changchun University of Chinese Medicine,Changchun Jilin 130117,China;Department of Infectious Diseases,Jilin Province People′s Hospital,Changchun Jinlin 130021,China;Precision Molecular Medicine Center,Jilin Province People′s Hospital,Changchun Jinlin 130021,China)
出处
《中国卫生标准管理》
2025年第18期160-165,共6页
China Health Standard Management
基金
2024年度吉林省中医药科技项目(2024123)。
关键词
核因子-ΚB信号通路
Toll样受体4
百合皂苷
人支气管肺上皮细胞
肺部炎症模型
脂多糖
nuclear factor-κB signaling pathway
Toll-like receptor 4
lilium saponins
human bronchopulmonary epithelial cells
pulmonary inflammation model
lipopolysaccharide
