睡莲花总多酚对脂多糖诱导A549细胞炎性损伤的保护作用研究  

Protective effects of total polyphenols from Nymphaea candida on lipopolysaccharide induced inflammatory injury in A549 cells

作　　者：丁宛婷 乌里盼·托乎达阿里 孙媛 姚雨含 李晨阳 赵军[1,2] DING Wan-Ting;WULIPAN Tuo-Hu-Da-A-Li;SUN Yuan;YAO Yu-Han;LI Chen-Yang;ZHAO Jun(School of Pharmacy,Xinjiang Medical University,Urumqi 830011,China;Xinjiang Key Laboratory for Uighur Medicine,Institute of Materia Medica of Xinjiang,Urumqi 830004,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]新疆药物研究所维吾尔药重点实验室,乌鲁木齐830004

出　　处：《食品安全质量检测学报》2025年第7期190-198,共9页Journal of Food Safety and Quality

基　　金：新疆维吾尔自治区自然科学基金重点项目(2021D01D14)。

摘　　要：目的研究睡莲花总多酚(total polyphenols from Nymphaea candid,NCTP)对脂多糖(lipopolysaccharide,LPS)诱导肺泡上皮细胞A549炎性损伤的保护作用。方法采用福林酚法测定NCTP中的多酚含量,高效液相色谱法(high performance liquid chromatography,HPLC)法测定其特征性成分含量。以1.0μg/mL LPS干预A549细胞12 h造成炎性损伤模型,CCK-8(cell counting kit-8)法检测细胞毒性,以筛选出合适的NCTP干预浓度;进一步将细胞分为空白组、脂多糖模型组、阳性对照地塞米松(0.039μg/mL dexamethasone,DXM)组、NCTP低中高剂量(6.0、15.0、30.0μg/mL)组。生化法检测一氧化氮(nitric oxide,NO)及前列素E2(prostaglandin E2,PGE2)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽(glutathione,GSH)活性;酶联免疫法(enzyme-linked immunosorbent assay,ELISA)检测白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;蛋白免疫印迹(Western Blot)法检测核转录因子E2相关因子2/血红素加氧酶-1(nuclear factor erythroid-2-related factor 2/heme oxygenase 1,Nrf2/HO-1)信号通路的相关蛋白[Kelch样ECH结合蛋白1(Kelch-like ECH-associating protein 1,Keap1)、核因子(红细胞衍生2)相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、血红素加氧酶1(heme oxygenase-1,HO-1)]表达。结果NCTP中多酚含量为(90.15±0.17)%,所含成分睡莲酚、鞣花酸和烟花苷含量分别为(28.05±0.02)%、(2.15±0.03)%和(3.50±0.01)%,CCK-8法筛选出NCTP的干预质量浓度为6.0、15.0、30.0μg/mL。LPS刺激A549细胞后,细胞内炎症介质NO、PGE2和炎症因子IL-1β、IL-6、TNF-α表达显著升高(P<0.01),说明A549细胞炎性损伤模型成功构建。与模型组比较,NCTP能显著抑制LPS诱导A549细胞上清中NO、PGE2、IL-1β、IL-6、TNF-α、MDA水平,明显提高细胞中SOD和GSH活力。与模型组相比,NCTP还能显著增加受损细Objective To study protective effects of total polyphenols from Nymphaea candida(NCTP)on lipopolysaccharide(LPS)induced inflammatory injury in A549 cells.Methods Polyphenol content in NCTP was determined by Folin-Ciocalte method,and its characteristic components were detected by high performance liquid chromatography(HPLC).A549 cells were treated with 1.0μg/mL LPS for 12 h to induce inflammatory damage model,and cell counting kit-8(CCK-8)method was used to screen out the optimal intervention concentration of NCTP.The experiment was divided into control group,lipopolysaccharide model group,dexamethasone(DXM 0.039μg/mL)group,and NCTP(6.0,15.0,30.0μg/mL)groups.The levels of nitric oxide(NO),prostaglandin E2(PGE2),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione(GSH)were detected by biochemical method.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in A549 cells.The expression of related proteins[Kelch-like ECH-associating protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1)]in nuclear factor erythroid-2-related factor 2/heme oxygenase 1(Nrf2/HO-1)signaling pathway was detected by Western Blot.Results The content of total polyphenols in NCTP was(90.15±0.17)%,and the content of isostrictiniin,ellagic acid and nicotiflorin was(28.05±0.02)%,(2.15±0.03)%and(3.50±0.01)%,respectively.The mass concentrations of NCTP screened by CCK-8 were 6.0,15.0,30.0μg/mL.After LPS stimulation of A549 cells,the expressions of intracellular inflammatory mediators NO,PGE2 and inflammatory factors IL-1β,IL-6 and TNF-αwere significantly increased(P<0.01),indicating that the inflammatory damage model of A549 cells was successfully constructed.Compared with the model group,NCTP significantly inhibited the expression of NO,PGE2,IL-1β,IL-6,TNF-αand MDA and increased the activities of SOD and GSH in LPS-induced A549 cells.Compared with the model group,NCTP also significantly incre

关 键 词：睡莲花 多酚 A549细胞 急性肺损伤 抗氧化作用 核转录因子E2相关因子2/血红素加氧酶-1通路 

分 类 号：R28[医药卫生—中药学]