Effects of baicalin on expression of mucin 5AC and cystic fibrosis transmembrane conductance regulator in airway epithelium through NF-κB signaling pathway

Yan ZHANG; Caili ZHANG; Yan LIU; Yulong CHEN; Mengwei PENG; Yazhou SANG; Yaosong WU

doi:10.16016/j.2097-0927.202109016

您当前的位置：

首页 >

文章列表页 >

Effects of baicalin on expression of mucin 5AC and cystic fibrosis transmembrane conductance regulator in airway epithelium through NF-κB signaling pathway

Basic Medicine | 更新时间：2026-01-09

- Effects of baicalin on expression of mucin 5AC and cystic fibrosis transmembrane conductance regulator in airway epithelium through NF-κB signaling pathway
- Journal of Army Medical University Vol. 44, Issue 5, Pages: 441-447(2022)
- 作者机构：
  
  1.450046 郑州, 河南中医药大学中医药科学院，河南省中医方证信号传导重点实验室/河南省中医方证信号传导国际联合重点实验室
  2.450046 郑州, 河南中医药大学中医药科学院，河南省中西医结合防治脑认知疾病工程技术研究中心
  3.450046 郑州, 河南中医药大学中医药科学院，郑州市中西医结合防治脑认知疾病重点实验室
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars of China(81503490);the Tackling Project of Science and Technology Plan of Henan Province(192102310160)
- DOI：10.16016/j.2097-0927.202109016
  CLC： R285.5;R322.34;R394.6
- Received：02 September 2021，
  
  Revised：2021-11-03，
  
  Published：15 March 2022
- 稿件说明：
移动端阅览
Yan ZHANG, Caili ZHANG, Yan LIU, et al. Effects of baicalin on expression of mucin 5AC and cystic fibrosis transmembrane conductance regulator in airway epithelium through NF-κB signaling pathway[J]. Journal of Army Medical University, 2022, 44(5): 441-447.
DOI：

Yan ZHANG, Caili ZHANG, Yan LIU, et al. Effects of baicalin on expression of mucin 5AC and cystic fibrosis transmembrane conductance regulator in airway epithelium through NF-κB signaling pathway[J]. Journal of Army Medical University, 2022, 44(5): 441-447. DOI： 10.16016/j.2097-0927.202109016.

摘要

目的

基于核因子κB(nuclear factor kappa B，NF-κB)信号探讨黄芩苷调控气道上皮黏蛋白5AC(mucin 5AC

MUC5AC)和囊性纤维化跨膜传导调节因子(cystic fibrosis transmembrane conductance regulator，CFTR)的表达以改善病理性气道黏液的机制。

方法

采用MTT法检测不同浓度黄芩苷对H292细胞(人肺癌细胞)活性的影响并确定用于后续实验的黄芩苷浓度；将细胞分为空白对照组、模型组、NF-κB抑制剂组(PDTC组，100 μmol/L)和黄芩苷组(200 μg/mL)。采用IL-1β诱导H292细胞建立体外黏液高分泌炎症细胞模型，再以相应浓度的药物干预24 h后，采用ELISA方法检测各组细胞上清中MUC5AC、IL-8、TNF-α的含量，检测各组细胞中MUC5AC和CFTR蛋白的含量；采用实时荧光定量PCR检测各组细胞中MUC5AC和CFTR mRNA的表达；采用Western blot检测各组细胞中p65和IκB的表达。

结果

MTT检测结果显示，当黄芩苷浓度为200 μg/mL时，细胞活性在80%以上，可作为后续实验的安全浓度。与空白对照组比较，模型组细胞培养上清及细胞质中MUC5AC分泌水平及IL-8、TNF-α、p65、MUC5AC mRNA表达均明显升高(

0.05)，IκB蛋白表达、CFTR蛋白含量、CFTR mRNA的表达显著降低(

0.05)；与模型组比较，PDTC组细胞培养上清中MUC5AC分泌水平，IL-8、TNF-α、p65蛋白表达及MUC5AC mRNA表达显著降低(

0.05)，细胞质中MUC5AC水平、CFTR蛋白含量、IκB蛋白表达、CFTR mRNA的表达显著升高(

0.05)；与模型组比较，黄芩苷组细胞培养上清中MUC5AC分泌水平、IL-8和TNF-α分泌水平、p65蛋白表达降低(

0.01)，CFTR蛋白含量、IκB蛋白表达、CFTR mRNA表达均显著增加(

0.01)，而MUC5AC mRNA表达差异无统计学意义。

结论

黄芩苷通过抑制NF-κB活性，减少MUC5AC的分泌，增加CFTR的表达，减轻气道炎症。

Abstract

Objective

To investigate the mechanism of baicalin in improving pathological airway mucus by regulating the expression of airway epithelial mucus regulators mucin 5AC (MUC5AC) and cystic fibrosis transmembrane conductance regulator (CFTR) through nuclear factor κB (NF-κB) signaling pathway.

Methods

MTT assay was used to detect the effects of different concentrations of baicalin on the viability of human lung cancer H292 cells in order to determine the proper dose of baicalin for subsequent experiments. After H292 cells were induced by IL-1β to establish

in vitro

inflammatory cell model of mucus hypersecretion

the cells were divided into blank control group

model group

NF-κB inhibitor group (PDTC group

100 μmol/L) and baicalin group (200 μg/mL). After 24 h of drug intervention

the contents of MUC5AC

IL-8

and TNF-α in the culture supernatant and the levels of MUC5AC and CFTR in the cytoplasm of each group were detected by ELISA. Real-time fluorescent quantitative PCR was adopted to detect the mRNA expression of MUC5AC and CFTR. Western blotting was performed to test the expression of p65 and IκB (inhibitor of NF-κB).

Results

The results of MTT assay showed that 200 μg/mL was a safe concentration of baicalin for subsequent experiments

with cell viability larger than 80%. As compared with the blank control group

the protein expression of MUC5AC

IL-8

TNF-α

p65 and the mRNA level of MUC5AC in culture supernatant and cytoplasm were all significantly increased in the model group (

0.05)

while the expression of IκB and CFTR proteins as well as the CFTR mRNA level were remarkably decreased (

0.05). When compared with the model group

the PDTC group presented lower level of MUC5AC secretion

declined protein levels of IL-8

TNF-α and p65

as well as MUC5AC mRNA in the culture supernatant (

0.05)

while higher protein expression of MUC5AC

CFTR and IκB in the cytoplasm

along with elevated mR

NA level of CFTR (

0.05). In terms of the baicalin group

the secretion levels of MUC5AC

IL-8 and TNF-α were greatly diminished in culture supernatant (

0.01) as compared with the model group

whereas the expression of IκB as well as the protein and mRNA levels of CFTR notably improved (

0.01)

and there was no significant difference in MUC5AC mRNA expression.

Conclusion

Baicalin reduces MUC5AC secretion and increases CFTR expression by inhibiting the activity of NF-κB

and thus attenuates airway inflammation.

关键词

Keywords

references

贾善学, 王洪星, 刘桂华, 等. RP-HPLC法测定礞石滚痰丸中黄芩苷的含量[J]. 中国药事, 2003, 17(3): 169-170.

JI A S X, WANG H X, LIU G H, et al . Content determination of baicaline in Mengshiguntan Pill by RP-HPLC[J ] . Chin Pharm Aff, 2003, 17(3): 169-170.

张浚, 张昌斌, 王锦绣. 反相高效液相色谱法测定竹沥达痰丸中黄芩苷的含量[J]. 时珍国医国药, 2008, 19(8): 1977-1978.

ZHANG J, ZHANG C B, WANG J X. Determination of baicalin in zhulidatan pills by HPLC[J]. Lishizhen Med Mater Med Res, 2008, 19(8): 1977-1978.

陆丽娜, 董加花, 左艳华, 等. RP-HPLC法同时测定清肺化痰丸中4种成分[J]. 中成药, 2016, 38(9): 1956-1959.

LU L N, DONG J H, ZUO Y H, et al . Simultaneous determination of four constituents in Qinfei Huatan Pills by RP-HPLC[J ] . Chin Tradit Pat Med, 2016, 38(9): 1956-1959.

屈飞, 吴汉松, 邱沙英, 等. 清肺化痰汤通过抑制EGFR通路下调MUC5AC的表达治疗COPD的研究[J]. 中药药理与临床, 2020, 36(4): 69-75. DOI: 10.13412/j.cnki.zyyl.2020.04.003.

QU F, WU H S, QIU S Y, et al . Treatment for COPD with qingfei huatan decoction by down regulating the expression of MUC5AC through EGFR pathway[J ] . Pharmacol Clin Chin Mater Med, 2020, 36(4): 69-75. DOI: 10.13412/j.cnki.zyyl.2020.04.003.

杨勤军, 王心恒, 杨程, 等. 麻芍平喘汤对哮喘大鼠气道重塑和气道炎症及肺组织TGF-β1、Muc5AC蛋白表达的影响[J]. 中药药理与临床, 2020, 36(5): 154-159, 64. DOI: 10.13412/j.cnki.zyyl.2020.05.017.

YANG Q J, WANG X H, YANG C, et al . Effects of Mashao Pingchuan decoction on airway remodeling, airway inflammation, expressions of TGF-β1 and Muc5AC in lung tissues of asthmatic rats[J ] . Pharmacol Clin Chin Mater Med, 2020, 36(5): 154-159, 64. DOI: 10.13412/j.cnki.zyyl.2020.05.017.

LILLEHOJ E P, KATO K, LU W, et al . Cellular and molecular biology of airway mucins[J ] . Int Rev Cell Mol Biol, 2013, 303: 139-202. DOI: 10.1016/b978-0-12-407697-6.00004-0.

COLLAWN J F, MATALON S. CFTR and lung homeostasis[J]. Am J Physiol Lung Cell Mol Physiol, 2014, 307(12): L917-L923. DOI: 10.1152/ajplung.00326.2014.

HAYDEN M S, GHOSH S. Shared principles in NF-kappaB signaling[J]. Cell, 2008, 132(3): 344-362. DOI: 10.1016/j.cell.2008.01.020.

RAMACHANDRAN S, KARP PH, OSTERHAUS S R, et al . Post-transcriptional regulation of cystic fibrosis transmembrane conductance regulator expression and function by microRNAs[J ] . Am J Respir Cell Mol Biol, 2013, 49(4): 544-551. DOI: 10.1165/rcmb.2012-0430OC.

刘书宇, 王树鹏. 基于NF-κB信号通路探讨麻黄细辛附子汤对变应性鼻炎大鼠MUC5AC和MUC5B表达影响[J]. 辽宁中医药大学学报, 2019, 21(12): 41-43. DOI: 10.13194/j.issn.1673-842x.2019.12.010.

LIU S Y, WANG S P. Exploring the effect of Mahuang Xixin Fuzi decoction on the expression of MUC5AC and MUC5B in rats with allergic rhinitis based on NF-kappa B signaling pathway[J]. J Liaoning Univ Tradit Chin Med, 2019, 21(12): 41-43. DOI: 10.13194/j.issn.1673-842x.2019.12.010.

刘立涛. 清热解毒类中药活性成分治疗慢性阻塞性肺病模型的筛选及作用机制探讨[D]. 吉首: 吉首大学, 2020.

LIU L T. Screening of the active components of Chinese medicine for clearing away heat and detoxification in the treatment of chronic obstructive pulmonary disease and its mechanism[D]. Jishou, China: Jishou University, 2020.

张培旭. 基于NF-κB信号通路研究小青龙汤对肺癌H292细胞分泌MUC5AC的调控机制[D]. 郑州: 河南中医药大学, 2018.

ZHANG P X. Study on the regulation mechanism of xiaoqinglong decoction on MUC5AC secretion in lung cancer H292 cells based on NF-κB signaling pathway[D]. Zhengzhou: Henan University of Chinese Medicine, 2018.

LEE E J, SONG K J, KWON J H, et al . Chronic cholesterol depletion by lovastatin suppresses MUC5AC gene expression in human airway epithelial cells[J ] . Am J Rhinol Allergy, 2014, 28(3): e125-e129. DOI: 10.2500/ajra.2014.28.4037.

蒋时红, 贺倩文, 张培旭, 等. 基于核转录因子-κB信号通路研究小青龙汤对肺癌H292细胞分泌黏蛋白5AC的调控机制[J]. 中国中西医结合急救杂志, 2020, 27(6): 733-737. DOI: 10.3969/j.issn.1008-9691.2020.06.025.

JIANG S H, HE Q W, ZHANG P X, et al . Regulatory mechanism of Xiaoqinglong Decoction on mucin 5AC secretion in lung cancer H292 cells based on nuclear transcription factor-κB signaling pathway[J ] . Chin J TCM WM Crit Care, 2020, 27(6): 733-737.

CAO Y G, MAO X Y, SUN C Y, et al . Baicalin attenuates global cerebral ischemia/reperfusion injury in gerbils via anti-oxidative and anti-apoptotic pathways[J ] . Brain Res Bull, 2011, 85(6): 396-402. DOI: 10.1016/j.brainresbull.2011.05.002.

张淑华, 官学苹, 阮奇军, 等. 黄芩苷对人结肠癌SW480细胞增殖和凋亡的影响及机制研究[J]. 现代中西医结合杂志, 2021, 30(21): 2299-2304.

ZHANG S H, GUAN X P, RUAN Q J, et al . Study on the effect of baicalin on proliferation and apoptosis of human colon cancer SW480 cells and its mechanism[J ] . Mod J Integr Tradit Chin West Med, 2021, 30(21): 2299-2304.

辛文妤, 宋俊科, 何国荣, 等. 黄芩素和黄芩苷的药理作用及机制研究进展[J]. 中国新药杂志, 2013, 22(6): 647-653, 659.

XIN W Y, SONG J K, HE G R, et al . Progress in pharmacological study and the underlying mechanism of baicalein and baicalin[J ] . Chin J New Drugs, 2013, 22(6): 647-653, 659.

LAPPALAINEN U, WHITSETT J A, WERT S E, et al . Interleukin-1beta causes pulmonary inflammation, emphysema, and airway remodeling in the adult murine lung[J ] . Am J Respir Cell Mol Biol, 2005, 32(4): 311-318. DOI: 10.1165/rcmb.2004-0309OC.

KIM Y D, JEON J Y, WOO H J, et al . Interleukin-1beta induces MUC 2 gene expression and mucin secretion via activation of PKC-MEK/ERK, and PI3K in human airway epithelial cells[J ] . J Korean Med Sci, 2002, 17(6): 765-771. DOI: 10.3346/jkms.2002.17.6.765..

李亚, 王洋, 毛静, 等. 补肺益肾方调控NF-κB信号通路对COPD大鼠炎性反应的影响[J]. 中华中医药杂志, 2021, 36(6): 3171-3175.

LI Y, WANG Y, MAO J, et al . Effects of Bufei Yishen Formula on lung inflammation via NF-κB signaling pathway in rats with COPD[J ] . China J Tradit Chin Med Pharm, 2021, 36(6): 3171-3175.

周必英, 周泠. 黄芩苷对卡氏肺孢子虫肺炎大鼠TNF-α和IL-8影响[J]. 中国公共卫生, 2009, 25(10): 1250-1251.

ZHOU B Y, ZHOU L. Influence of baicalin on tumour necrosis factor-α and interleukin-8 in rats infected with Pneumocystis carinii pneumonia[J]. Chin J Public Heal, 2009, 25(10): 1250-1251.

BROIDE D H, LAWRENCE T, DOHERTY T, et al . Allergen-induced peribronchial fibrosis and mucus production mediated by IkappaB kinase beta-dependent genesin airway epithelium[J ] . Proc Natl Acad Sci USA, 2005, 102(49): 17723-17728. DOI: 10.1073/pnas.0509235102.

SIKDER M A, LEE H J, MIA M Z, et al . Inhibition of TNF-α-induced MUC5AC mucin gene expression and production by wogonin through the inactivation of NF-κB signaling in airway epithelial cells[J ] . Phytother Res, 2014, 28(1): 62-68. DOI: 10.1002/ptr.4954.

XU X L, HUANG H M, YIN X Y, et al . Effect of Lentivirus-mediated CFTR overexpression on oxidative stress injury and inflammatory response in the lung tissue of COPD mouse model[J ] . Biosci Rep, 2020, 40(1): BSR20193667. DOI: 10.1042/BSR20193667.

WU S, LI H, YU L, et al . IL-1β upregulates Muc5ac expression via NF-κB-induced HIF-1α in asthma[J ] . Immunol Lett, 2017, 192: 20-26. DOI: 10.1016/j.imlet.2017.10.006.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Celastrol attenuates sodium oxalate-induced acute kidney injury and crystal deposition by inhibiting NF-κB

Naringin improves lipopolysaccharide-induced depression-like behaviors in mice

LncRNA Gm4419 regulates inflammatory cytokines expression via NF-κB signaling pathway in mesangial cells under high glucose condition

1,25(OH)2D3 inhibits osteoclastogenesis by IκB/NF-κB signaling pathway

Role of NF-κB signaling pathway in treatment of intrauterine adhesions by human amniotic mesenchymal stem cells

Related Author

LIU Yiheng

ZHENG Quanyou

ZHANG Wanyuan

YANG Chenhao

CHEN Siyu

LIN Wenbiao

ZHAO Siyu

XU Guilian

Related Institution

Urinary Nephropathy Center， the Second Affiliated Hospital of Chongqing Medical University

Department of Immunology， College of Basic Medical Sciences

Department of Nephrology and Urology， No. Hospital958， First Affiliated Hospital， Army Medical University （Third Military Medical University）

Department of Military Cognitive Psychology, Faculty of Medical Psychology, Army Medical University (Third Military Medical University)

Department of Cell Biology and Medical Genetics, College of Basic Medical Sciences, Chongqing Medical University

⁰