Page 34 - 《中国药房》2022年12期
P. 34
牛蒡子苷减轻脂多糖诱导人鼻咽上皮细胞炎症损伤的作用 Δ
*
聂发龙 ,赵显芳,朱紫陌,江自鲜,代 蓉,李秀芳(云南中医药大学中药学院,昆明 650500)
#
中图分类号 R965 文献标志码 A 文章编号 1001-0408(2022)12-1436-06
DOI 10.6039/j.issn.1001-0408.2022.12.05
摘 要 目的 探讨牛蒡子苷(ARC)减轻脂多糖(LPS)诱导的人鼻咽上皮细胞 NP-69 炎症损伤的作用。方法 采用 MTS 法测定
0.000 1、0.001、0.01、0.1、1.0、10 μmol/L ARC 作用 24 h 对 NP-69 细胞增殖的影响。采用划痕实验分别检测经 0.01、0.1、1.0 μmol/L
ARC 作用 24 h 和经 0.01、0.1、1.0 μmol/L ARC 预处理 24 h+1.0 μg/mL LPS 刺激 24 h 后 NP-69 细胞的迁移情况。以 LPS 刺激建立
NP-69细胞炎症损伤模型,检测上清液中一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β水平以及细胞中胞
质紧密连接蛋白1(ZO-1)、β-防御素3(BD3)、Janus激酶1(JAK1)、信号转导及转录活化因子3(STAT3)mRNA和蛋白的相对表达
量。结果 与正常组比较,0.000 1、0.001、0.01、0.1、1.0、10 μmol/L ARC 作用 24 h 对 NP-69 细胞增殖均无影响(P>0.05)。0.1、
1.0 μmol/L ARC 均可显著提高细胞的迁移率(P<0.05)。对 LPS 刺激 NP-69 细胞造成的炎症损伤,ARC(1.0 μmol/L)能显著减少
NO、TNF-α、IL-6的释放(P<0.05),同时能显著上调ZO-1、BD3 mRNA和蛋白的表达并下调STAT3 mRNA和蛋白的表达(P<0.01
或P<0.05)。结论 ARC具有明显减轻LPS致NP-69细胞炎症损伤的作用,并可增强炎症环境下鼻黏膜上皮屏障的物理及免疫防
御功能;上述作用可能与抑制IL-6/JAK1/STAT3信号通路相关。
关键词 牛蒡子苷;脂多糖;NP-69细胞;炎症反应;鼻黏膜上皮屏障
Effects of arctiin relieving inflammatory injury of human nasopharyngeal epithelial cells induced by
lipopolysaccharide
NIE Falong,ZHAO Xianfang,ZHU Zimo,JIANG Zixian,DAI Rong,LI Xiufang(College of Traditional Chinese
Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)
ABSTRACT OBJECTIVE To investigate the effect of arctiin(ARC)relieving lipopolysaccharide(LPS)induced inflammatory
injury of human nasopharyngeal epithelial cells NP-69. METHODS The effects of 24 h treatment of 0.000 1,0.001,0.01,0.1,
1.0,10 μmol/L ARC on the proliferation of NP-69 were determined by MTS method. After 0.01,0.1,and 1.0 μmol/L ARC was
applied to NP-69 for 24 h and NP-69 was pre-treated with 0.01,0.1 and 1.0 μmol/L ARC for 24 h,and then stimulated with 1.0
μg/mL LPS for 24 h,scratch tests were used to detect cell migration in both experiments. LPS stimulated NP-69 to establish an
inflammation injury model. The levels of nitric oxide(NO),tumor necrosis factor α(TNF-α)interleukin-6(IL-6),and IL-1β in
cell supernatants were detected,and mRNA and protein expression of zonula oecludens protein 1(ZO-1),β-defensin 3(BD3),
Janus kinase 1 (JAK1),signal transducer and activator of transcription 3 (STAT3) in cell supernatant were also detected.
RESULTS Compared with normal group,0.000 1,0.001,0.01,0.1,1.0,10 μmol/L ARC had no effect on the proliferation of
NP-69 after 24 h treatment(P>0.05). ARC(0.1,1.0 μmol/L)could significantly promote the rate of cell migration(P<0.05).
For the inflammatory injure of NP-69 cells stimulated by LPS,ARC(1.0 μmol/L)could significantly reduce the release of NO,
TNF-α and IL-6(P<0.05),significantly increased mRNA and protein expression of ZO-1 and BD3 but decreased mRNA and
protein expression of STAT3(P<0.01 or P<0.05). CONCLUSIONS ARC has the effect of reducing the inflammatory injury of
NP-69 cells induced by LPS,promoting the physical and immune defense ability of the nasal mucosa epithelial barrierunder
inflammatory environment. The mechanism of action may be related to inhibiting IL-6/JAK1/STAT3 signaling pathway.
KEYWORDS arctiin;lipopolysaccharide;NP-69;inflammation;nasopharyngeal epithelial barrier
上 呼 吸 道 感 染(upper respiratory tract infection, 为与外界直接相连的器官,鼻咽上皮细胞在呼吸时会暴
URTI)是鼻、咽、喉急性炎症的总称,是危害人类健康的 露于空气当中,容易受到病毒、细菌、真菌等病原微生物
常见疾病之一,且大多由病毒和细菌感染所致 [1-2] 。作 攻击,作为机体防御URTI的首道屏障,鼻咽上皮细胞对
[3]
Δ 基金项目:国家自然科学基金资助项目(No.81860724);云南省 维持上呼吸道的稳定具有重要作用 。病原微生物致感
科技厅科技计划项目[No.202001AZ070001-001、No.2019FF002(-054)] 染或炎症信号增强等多种因素的作用会导致鼻黏膜上
*硕士研究生。研究方向:中药药理与应用。E-mail:1169716958@
皮细胞受到损伤并发生功能紊乱,进而易使外界病原体
qq.com
或机会致病菌进入黏膜,最终引发URTI 。牛蒡子为菊
[4]
# 通信作者:教授,硕士生导师,博士。研究方向:中药药理与应
用。E-mail:sofinelxf@163.com 科植物牛蒡Arctium lappa L.的干燥成熟果实,是中医临
·1436 · China Pharmacy 2022 Vol. 33 No. 12 中国药房 2022年第33卷第12期