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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
Received:07 December 2022,
Published Online:25 March 2023,
Published:05 December 2023
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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.
目的
2
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-
κ
B通路活性的影响。
方法
2
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
β
、IL-4、IL-13、转化生长因子(TGF)-
β
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-
κ
B(NF-
κ
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-
κ
B、磷酸化NF-
κ
B(p-NF-
κ
B)及核蛋白NF-
κ
B(NEs-NF-
κ
B)蛋白相对表达量,免疫荧光法检测NF-
κ
B核转位水平。
结果
2
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
β
、IL-4、IL-13、TGF-
β
含量明显增加(
P
<
0.05,
P
<
0.01),TLR4、MyD88、NF-
κ
B、p-NF-
κ
B、NEs-NF-
κ
B、IL-4表达水平显著升高(
P
<
0.01),NF-
κ
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
β
、IL-4、IL-13、TGF-
β
含量及TLR4、MyD88、NF-
κ
B、p-NF-
κ
B、NEs-NF-
κ
B、IL-4表达水平明显降低(
P
<
0.05,
P
<
0.01),NF-
κ
B入核明显减少。
结论
2
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-
κ
B信号通路活性有关。
Objective
2
To observe the effects of
Dendrobium
polysaccharides on the secretion of inflammatory cytokines and Toll-like receptor 4 (TLR4)/nuclear factor (NF)-
κ
B pathway in 16HBE cells exposed to cigarette smoke extract (CSE).
Method
2
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
β
, IL-4, IL-13, and transforming growth factor (TGF)-
β
in cell culture supernatants. Real-time PCR was carried out to determine the mRNA levels of Toll-like receptor 4 (TLR4), nuclear factor-
κ
B (NF-
κ
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-
κ
B, phosphorylated nuclear factor-
κ
B (p-NF-
κ
B), and nucleoproteins nuclear factor-
κ
B (NEs-NF-
κ
B). The immunofluorescence assay was employed to measure the nuclear translocation of NF-
κ
B.
Result
2
Compared with the control group, the CSE group showed elevated levels of IL-8, IL-1
β
, IL-4, IL-13, and TGF-
β
in
the cell culture supernatants (
P
<
0.05,
P
<
0.01), up-regulated expression levels of TLR4, MyD88, NF-
κ
B, p-NF-
κ
B, NEs-NF-
κ
B, and IL-4 (
P
<
0.01), and significant nuclear translocation of NF-
κ
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
β
, IL-4, IL-13, and TGF-
β
, down-regulated the expression of TLR4, MyD88, NF-
κ
B, p-NF-
κ
B, NEs-NF-
κ
B, and IL-4 (
P
<
0.05,
P
<
0.01), and reduced the nuclear translocation of NF-
κ
B.
Conclusion
2
Dendrobium polysaccharides showed significant protective effects on the 16HBE cells exposed to CSE by inhibiting the TLR4/NF-
κ
B signaling pathway.
LABAKI W W , ROSENBERG S R . Chronic obstructive pulmonary disease [J]. Ann Intern Med , 2020 , 173 ( 3 ): ITC17 - ITC32 .
WANG C , XU J , YANG L , et al . Prevalence and risk factors of chronic obstructive pulmonary disease in China (the China Pulmonary Health [CPH] study):A national cross-sectional study [J]. Lancet , 2018 , 391 ( 10131 ): 1706 - 1717 .
LI Y , ZHAO J , SHAO H , et al . Preventive effect of total flavonoids of Trollius altaicus on a chronic obstructive pulmonary disease rat model based on the TLR4/NF- κ B pathway [J]. Ann Transl Med , 2022 , 10 ( 4 ): 222 .
LIAO Y N , HU W L , CHEN H J , et al . The use of chinese herbal medicine in the treatment of chronic obstructive pulmonary disease (COPD) [J]. Am J Chin Med , 2017 , 45 ( 2 ): 225 - 238 .
ZHU Y , OUYANG Z , DU H , et al . New opportunities and challenges of natural products research:When target identification meets single-cell multiomics [J]. Acta Pharm Sin B , 2022 , 12 ( 11 ): 4011 - 4039 .
HE Y , LI L , CHANG H , et al . Research progress on extraction,purification,structure and biological activity of polysaccharides [J]. Front Nutr , 2022 , 9 : 965073 .
LIANG J , WU Y , YUAN H , et al . Dendrobium officinale polysaccharides attenuate learning and memory disabilities via anti-oxidant and anti-inflammatory actions [J]. Int J Biol Macromol , 2019 , 126 : 414 - 426 .
ZHAO Y , HUANG H X , TANG S Y , et al . RNA-Seq analysis reveals polysaccharides inhibit precancerous lesions of gastric cancer through PER3 and AQP4 [J]. Evid Based Complement Alternat Med , 2021 , 2021 : 3036504 .
XIANG L , STEPHEN SZE C W , NG T B , et al . Polysaccharides of Dendrobium officinale inhibit TNF- α -induced apoptosis in A-253 cell line [J]. Inflamm Res , 2013 , 62 ( 3 ): 313 - 324 .
HUANG K , LI Y , TAO S , et al . Purification,characterization and biological activity of polysaccharides from Dendrobium officinale [J]. Molecules , 2016 , doi: 10.3390/MOLECULES2106-0701 http://dx.doi.org/10.3390/MOLECULES2106-0701 .
WEN Y , XIAO H , LIU Y , et al . Polysaccharides from Dendrobium officinale ameliorate colitis-induced lung injury via inhibiting inflammation and oxidative stress [J]. Chem Biol Interact , 2021 , 347 : 109615 .
CHEN J , LU J , WANG B , et al . Polysaccharides from Dendrobium officinale inhibit bleomycin-induced pulmonary fibrosis via the TGF- β 1 -Smad2/3 axis [J]. Int J Biol Macromol , 2018 , 118 ( Pt B ): 2163 - 2175 .
LIANG Y , DU R , CHEN R , et al . Therapeutic potential and mechanism of Dendrobium officinale polysaccharides on cigarette smoke-induced airway inflammation in rat [J]. Biomed Pharmacother , 2021 , 143 : 112101 .
DI VINCENZO S , HEIJINK I H , NOORDHOEK J A , et al . SIRT1/FoxO3 axis alteration leads to aberrant immune responses in bronchial epithelial cells [J]. J Cell Mol Med , 2018 , 22 ( 4 ): 2272 - 2282 .
YANG H , ZHANG L , WANG Q . MicroRNA-221-3p alleviates cell apoptosis and inflammatory response by targeting cyclin dependent kinase inhibitor 1B in chronic obstructive pulmonary disease [J]. Bioengineered , 2021 , 12 ( 1 ): 5705 - 5715 .
LIN F , LIAO C , ZHANG J , et al . Hydrogen sulfide inhibits bronchial epithelial cell epithelial mesenchymal transition through regulating endoplasm reticulum stress [J]. Front Mol Biosci , 2022 , 9 : 828766 .
刘振峰 , 刘建英 , 刘代顺 , 等 . 香烟烟雾提取物诱导呼吸道上皮细胞间质性转化的实验研究 [J]. 重庆医学 , 2017 , 46 ( 3 ): 318 - 321 .
MA H , ZHOU Z , CHEN L , et al . Anemoside B4 prevents chronic obstructive pulmonary disease through alleviating cigarette smoke-induced inflammatory response and airway epithelial hyperplasia [J]. Phytomedicine , 2022 , 107 : 154431 .
OU G , ZHU M , HUANG Y , et al . HSP60 regulates the cigarette smoke-induced activation of TLR4-NF- κ B-MyD88 signalling pathway and NLRP3 inflammasome [J]. Int Immunopharmacol , 2022 , 103 : 108445 .
FAN Y , ZHANG Z , YAO C , et al . Amurensin H,a derivative from resveratrol,ameliorates lipopolysaccharide/cigarette smoke-induced airway inflammation by blocking the Syk/NF- κ B pathway [J]. Front Pharmacol , 2019 , 10 : 1157 .
王梦文 , 郭艳 , 王重阳 , 等 . 化痰降气胶囊含药血清调控Nrf2/HDAC2改善16HBE细胞糖皮质激素抵抗 [J]. 中国临床药理学与治疗学 , 2021 , 26 ( 12 ): 1360 - 1369 .
SHEN Y , LU H , SONG G . MiR-221-3p and miR-92a-3p enhances smoking-induced inflammation in COPD [J]. J Clin Lab Anal , 2021 , 35 ( 7 ): e23857 .
屈飞 , 邱沙英 , 张明月 , 等 . 清肺化痰汤通过抑制JAK2/STAT3通路上调CFTR的表达治疗COPD [J]. 中国实验方剂学杂志 , 2020 , 26 ( 18 ): 1 - 7 .
LI Z , MAO X , LIU Q , et al . Functional variations of the TLR4 gene in association with chronic obstructive pulmonary disease and pulmonary tuberculosis [J]. BMC Pulm Med , 2019 , 19 ( 1 ): 184 .
WANG H , LI S , LI H , et al . Mechanism of probiotic VSL#3 inhibiting NF- κ B and TNF- α on Colitis through TLR4-NF- κ B signal pathway [J]. Iran J Public Health , 2019 , 48 ( 7 ): 1292 - 1300 .
YANG K , ZHAN L , LU T , et al . Dendrobium officinale polysaccharides protected against ethanol-induced acute liver injury in vivo and in vitro via the TLR4/NF- κ B signaling pathway [J]. Cytokine , 2020 , 130 : 155058 .
GUO L , QI J , DU D , et al . Current advances of polysaccharides in dermatology:A literature review [J]. Pharm Biol , 2020 , 58 ( 1 ): 664 - 673 .
尚立芝 , 李耀洋 , 季书 , 等 . 二陈汤加味通过HMGB1/RAGE/NF- κ B信号通路对COPD大鼠细支气管炎症的影响 [J]. 中国实验方剂学杂志 , 2023 , 29 ( 6 ): 44 - 54 .
LI Y , LU J , ZHANG Y , et al . Macrophages polarization and their role in chronic obstructive pulmonary disease [J]. Acta Physiol Sinica , 2019 , 71 ( 4 ): 604 - 612 .
WANG K , YANG X , WU Z , et al . Polysaccharide protected CCl-induced liver fibrosis through intestinal homeostasis and the LPS-TLR4-NF- κ B signaling pathway [J]. Front Pharmacol , 2020 , 11 : 240 .
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