Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-<italic style="font-style: italic">κ</italic>B Pathway

WANG Lei; WANG Yajuan; HE Zengyang; XU Fengqing; WANG Wenbin; WANG Tongsheng; SHU Junsheng

doi:10.13422/j.cnki.syfjx.20230805

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Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway

更新时间：2023-11-07

- Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 23, Pages: 64-71(2023)
- 作者机构：
  
  1.安徽中医药大学，合肥 230012
  2.安徽中烟工业有限责任公司技术中心，合肥 230088
  3.安徽中医药大学药学院，合肥 230012
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230805
  CLC： R2-0;R22;R285.5;R289;R33
- Published：05 December 2023，
  
  Published Online：25 March 2023，
  
  Received：07 December 2022，
- 稿件说明：
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王蕾,王雅娟,贺增洋等.基于TLR4/NF-κB通路探讨石斛多糖改善CSE诱导的人支气管上皮细胞炎性损伤[J].中国实验方剂学杂志,2023,29(23):64-71.

WANG Lei,WANG Yajuan,HE Zengyang,et al.Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):64-71.
王蕾,王雅娟,贺增洋等.基于TLR4/NF-κB通路探讨石斛多糖改善CSE诱导的人支气管上皮细胞炎性损伤[J].中国实验方剂学杂志,2023,29(23):64-71. DOI： 10.13422/j.cnki.syfjx.20230805.

WANG Lei,WANG Yajuan,HE Zengyang,et al.Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):64-71. DOI： 10.13422/j.cnki.syfjx.20230805.

摘要

目的

探讨石斛多糖对香烟烟雾提取物（CSE）损伤的人支气管上皮细胞（16HBE）的炎症因子分泌及TLR4/NF-

B通路活性的影响。

方法

体外培养16HBE细胞，设置空白组、CSE损伤组、CSE+石斛多糖组（100、200、400 mg·L

-1

），CCK-8法检测细胞活力，显微镜观察细胞状态，酶联免疫吸附测定法（ELISA）检测细胞培养上清中白细胞介素（IL）-8、IL-1

、IL-4、IL-13、转化生长因子（TGF）-

含量，实时荧光定量聚合酶链式反应（Real-time PCR）检测Toll样受体4（TLR4）、核转录因子-

B（NF-

B）及IL-4 mRNA的相对表达量，蛋白免疫印迹法（Western blot）检测白细胞介素-4受体（IL-4R）、TLR4、髓系分化初级反应蛋白88（MyD88）、NF-

B、磷酸化NF-

B（p-NF-

B）及核蛋白NF-

B（NEs-NF-

B）蛋白相对表达量，免疫荧光法检测NF-

B核转位水平。

结果

与空白组比较，CSE损伤组细胞存活率降低，细胞皱缩，细胞培养上清中IL-8、IL-1

、IL-4、IL-13、TGF-

含量明显增加（

0.05，

0.01），TLR4、MyD88、NF-

B、p-NF-

B、NEs-NF-

B、IL-4表达水平显著升高（

0.01），NF-

B入核现象明显；与CSE损伤组比较，石斛多糖各剂量组细胞存活率显著升高，细胞状态恢复，IL-8、IL-1

、IL-4、IL-13、TGF-

含量及TLR4、MyD88、NF-

B、p-NF-

B、NEs-NF-

B、IL-4表达水平明显降低（

0.05，

0.01），NF-

B入核明显减少。

结论

石斛多糖对CSE诱导的16HBE细胞损伤具有明显的保护作用，其机制可能与抑制TLR4/NF-

B信号通路活性有关。

Abstract

Objective

To observe the effects of

Dendrobium

polysaccharides on the secretion of inflammatory cytokines and Toll-like receptor 4 （TLR4）/nuclear factor （NF）-

B pathway in 16HBE cells exposed to cigarette smoke extract （CSE）.

Method

The 16HBE cells were classified into the control， CSE， and CSE+ Dendrobium polysaccharides （100， 200， 400 mg·L

-1

） groups. The cell-counting kit-8 （CCK-8） assay was employed to measure the cell viability， and a microscope was used to observe the cell morphology. The enzyme-linked immunosorbent assay was employed to measure the levels of interleukin （IL）-8， IL-1

， IL-4， IL-13， and transforming growth factor （TGF）-

in cell culture supernatants. Real-time PCR was carried out to determine the mRNA levels of Toll-like receptor 4 （TLR4）， nuclear factor-

B （NF-

B）， and IL-4. Western blot was employed to determine the protein levels of interleukin-4 receptor （IL-4R）， TLR4， myeloid differentiation primary response protein 88 （MyD88）， NF-

B， phosphorylated nuclear factor-

B （p-NF-

B）， and nucleoproteins nuclear factor-

B （NEs-NF-

B）. The immunofluorescence assay was employed to measure the nuclear translocation of NF-

Result

Compared with the control group， the CSE group showed elevated levels of IL-8， IL-1

， IL-4， IL-13， and TGF-

the cell culture supernatants （

0.05，

0.01）， up-regulated expression levels of TLR4， MyD88， NF-

B， p-NF-

B， NEs-NF-

B， and IL-4 （

0.01）， and significant nuclear translocation of NF-

B. Compared with the CSE group， Dendrobium polysaccharides increased the cell survival rate， recovered the cell activity， lowered the levels of IL-8， IL-1

， IL-4， IL-13， and TGF-

， down-regulated the expression of TLR4， MyD88， NF-

B， p-NF-

B， NEs-NF-

B， and IL-4 （

0.05，

0.01）， and reduced the nuclear translocation of NF-

Conclusion

Dendrobium polysaccharides showed significant protective effects on the 16HBE cells exposed to CSE by inhibiting the TLR4/NF-

B signaling pathway.

关键词

石斛多糖人支气管上皮细胞（16HBE）香烟烟雾提取物（CSE）炎症因子Toll样受体4（TLR4）核转录因子-κB（NF-κB）

Keywords

Dendrobium polysaccharides16HBE cellscigarette smoke extract （CSE）inflammatory cytokineToll-like receptor 4 （TLR4）nuclear factor-κB （NF-κB）

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Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway

DOI：10.13422/j.cnki.syfjx.20230805

摘要

Abstract

关键词

Keywords

references