首页|苦豆碱通过抑制TLR4/NF-κB/NLRP3通路改善香烟烟雾诱导的人支气管上皮细胞损伤

苦豆碱通过抑制TLR4/NF-κB/NLRP3通路改善香烟烟雾诱导的人支气管上皮细胞损伤

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目的 探究苦豆碱(Alo)对香烟烟雾诱导的人支气管上皮细胞损伤的作用及其可能的作用机制。方法 16HBE人支气管上皮细胞经100mL/L香烟烟雾提取物(CSE)和(50、100、200)μmol/L Alo共处理后，CCK-8法检测细胞活力，试剂盒检测乳酸脱氢酶(LDH)活性;原位末端转移酶标记技术(TUNEL)、Western blot法检测细胞凋亡，ELISA检测炎性因子水平;2'，7'-二氯二氢荧光素二乙酸酯(DCFH-DA)荧光探针和相关试剂盒检测氧化应激水平;Western blot法检测Toll样受体4(TLR4)/核因子κB(NF-κB)/含pyrin结构域核苷酸结合寡聚结构域样受体家族蛋白3(NLRP3)通路相关蛋白表达水平。16HBE细胞经100 mL/L CSE和200 μmol/L Alo共处理后，采用上述方法检测过表达TLR4对TLR4/NF-κB/NLRP3通路、细胞LDH活性、凋亡、炎症反应及氧化应激的影响。结果 CSE暴露可降低16HBE细胞活力，增加LDH释放和细胞凋亡，增强炎症反应和氧化应激水平，且激活TLR4/NF-κB/NLRP3通路;经Alo处理后，细胞活性升高，LDH释放减少、凋亡降低、炎症减轻、氧化应激水平下降，且TLR4/NF-κB/NLRP3通路失活;TLR4过表达可逆转Alo处理对CSE诱导的16HBE细胞损伤的保护作用。结论 Alo可通过抑制TLR4/NF-κB/NLRP3通路减轻CSE诱导的人支气管上皮细胞损伤。
Aloperine suppresses TLR4/NF-κB/NLRP3 signaling to ameliorate cigarette smoke-induced injury to human bronchial epithelial cells
Objective To explore the effects of aloperine(Alo)on cigarette smoke-induced injury in human bronchial epithelial cells and its potential mechanism.Methods After human bronchial epithelial 16HBE cells were co-treated by 100 mL/L cigarette smoke extract(CSE)and various concentrations(50,100 and 200 μmoVL)of Alo,cell viability was assessed using CCK-8 assay.Lactate dehydrogenase(LDH)activity was measured with a related kit.Cell apoptosis was evaluated using the terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay(TUNEL)and Western blot analysis.The levels of inflammatory factors were detected by ELISA.Oxidative stress levels were assessed using 2'7'-dichlorofluorescin diacetate(DCFH-DA)staining.The expression of Toll-like receptor 4(TLR4)/nuclear factor-kappaB(NF-κB)/NLR family pyrin domain containing 3(NLRP3)signaling-asscciated proteins was measured by Western blot analysis.After cells were co-treated with 100 mL/L CSE and 200 μmoVL Alo,the aforementioned assays were applied to evaluate the effects of TLR4 overexpression on the TLR4/NF-κB/NLRP3 signaling,LDH activity,apoptosis,inflammatory response and oxidative stress in cells.Results CSE exposure might inhibit 16HBE cell viability,increase LDH activity,apoptosis,inflammatory response and oxidative stress levels and activate TLR4/NF-κB/NLRP3 signaling.Treatment with Alo promoted cell viability,decreased LDH activity,cell apoptosis,inflammation and oxidative stress levels,and inactivated TLR4/NF-κB/NLRP3 signaling.Furthermore,TLR4 overexpression might reverse the protective role of Alo treatment in CSE-induced injury in 16HBE cells.Conclusion Alo may ameliorate CSE-induced injury in human bronchial epithelial cells via inhibiting TLR4/NF-κB/NLRP3 signaling.

bronchial epithelial cellscigarette smokealoperine(Alo)Toll-like receptor 4(TLR4)nuclear factor-kappaB(NF-κB)NLR family pyrin domain containing 3(NLRP3)

王慧、闫晓培、徐莉

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南京医科大学附属苏州医院/苏州市立医院呼吸与危重症医学科,江苏苏州 215000

支气管上皮细胞 香烟烟雾 苦豆碱 Toll样受体4(TLR4) 核因子κB(NF-κB) 含pyrin结构域核苷酸结合寡聚结构域样受体家族蛋白3(NLRP3)

南京医科大学科技发展基金

NMUB20210286

2024

10.13423/j.cnki.cjcmi.009790

细胞与分子免疫学杂志
中国免疫学会,第四军医大学

细胞与分子免疫学杂志

CSTPCD北大核心
影响因子：0.817
ISSN：1007-8738
年,卷(期)：2024.40(5)
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