Recombinant human CC16 inhibits senescence in human bronchial epithelial cells induced by cigarette smoke extract

Ting LI; Xiaoxue YANG; Rui GAO; Xinyang LI; Hailong WANG; Min PANG

doi:10.16016/j.2097-0927.202309135

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Recombinant human CC16 inhibits senescence in human bronchial epithelial cells induced by cigarette smoke extract

Basic Medicine | 更新时间：2026-01-09

- Recombinant human CC16 inhibits senescence in human bronchial epithelial cells induced by cigarette smoke extract
- Journal of Army Medical University Vol. 46, Issue 12, Pages: 1410-1416(2024)
- 作者机构：
  
  1.030001 太原，山西医科大学第一医院呼吸与危重症医学科，呼吸疾病防治与基础研究山西省重点实验室
  2.030600 山西晋中，山西医科大学基础医学研究中心
- 作者简介：
- 基金信息：
  
  the Research Foundation for Overseas Returners of Shanxi Province(2022-191);the Natural Science Fund of Shanxi Province(202103021223433);the "136" Special Fund Project of the First Hospital of Shanxi Medical University(2021-07)
- DOI：10.16016/j.2097-0927.202309135
  CLC： R322.34;R329.28;R977.6
- Received：24 September 2023，
  
  Revised：2023-12-06，
  
  Published：30 June 2024
- 稿件说明：
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Ting LI, Xiaoxue YANG, Rui GAO, et al. Recombinant human CC16 inhibits senescence in human bronchial epithelial cells induced by cigarette smoke extract[J]. Journal of Army Medical University, 2024, 46(12): 1410-1416.
DOI：

Ting LI, Xiaoxue YANG, Rui GAO, et al. Recombinant human CC16 inhibits senescence in human bronchial epithelial cells induced by cigarette smoke extract[J]. Journal of Army Medical University, 2024, 46(12): 1410-1416. DOI： 10.16016/j.2097-0927.202309135.

摘要

目的

探讨重组人CC16蛋白质(rhCC16)对香烟烟雾提取物(cigarette smoke extract

CSE)诱导的人支气管上皮细胞(human bronchial epithelial cells

HBECs)衰老的抑制作用及机制。

方法

CCK-8法检测CSE (0%、1%、2.5%、5%、7.5%和10%)及rhCC16(0、10、100、250和500 ng/mL)对HBECs细胞活力的影响; β-半乳糖苷酶染色试剂盒检测对照组、CSE组以及CSE+rhCC16组细胞β-半乳糖苷酶活性; 实时荧光定量PCR (qPCR

)检测各组细胞

16、

21 mRNA表达水平; Western blot检测各组细胞P16、P21、p-P53、P38、p-P38、ERK1/2、p-ERK1/2蛋白表达水平。

结果

与对照组相比

5% CSE刺激细胞72 h对HBECs细胞活力无显著影响; 500 ng/mL及以下浓度rhCC16对HBECs细胞活力无显著影响; 与对照组相比

5% CSE刺激HBECs细胞72 h致β-半乳糖苷酶染色阳性细胞率明显升高(

0.05)

且P16和P21蛋白和mRNA表达水平明显升高(

0.05)

p-P53、p-P38与p-ERK1/2蛋白表达量显著增加(

0.05);与CSE组相比

250 ng/mL rhCC16干预下

HBECs细胞β-半乳糖苷酶染色阳性细胞率显著下降(

0.05)

P16和P21蛋白和mRNA表达水平显著下降(

0.05)

p-P53、p-p38和p-ERK1/2的表达量降低(

0.05)。

结论

rhCC16蛋白质抑制香烟烟雾诱导的HBECs衰老

抑制P38 MAPK和ERK1/2信号通路可能是其作用机制。

Abstract

Objective

To investigate the effect and mechanism of recombinant human CC16 protein (rhCC16) on cigarette smoke extract (CSE)-induced senescence of human bronchial epithelial cells (HBECs).

Methods

CCK-8 assay was used to evaluate the cell viability in HBECs after treated with 0%

2.5%

7.5% and 10% CSE and/or 0

100

250 and 500 ng/mL rhCC16.β-galactosidase staining was used to observe the activity of the enzyme in control

CSE

and CSE+rhCC16 treated HBECs.The mRNA levels of

16 and

21 in above cell groups were detected by real-time fluorescence quantitative PCR (qPCR)

and the protein levels of P16

P21

p-P53

P38

p-P38

ERK1/2 and p-ERK1/2 were detected with Western blot in control group

CSE and CSE+rhCC16 groups.

Results

Compared with the control group

5% CSE stimulation for 72 h or the treatment of 500 ng/mL or lower dose of rhCC16 had no significant effect on cell viability of HBECs.Stimulation 5% CSE for 72 h resulted in significantly higher positive rate to β-galactosidase staining (

0.05)

elevated mRNA and protein levels of P16 and P21(

0.05)

and enhanced protein levels of p-P53

p-P38 and p-ERK1/2(

0.05) when compared with the control group.Compared with the simple CSE stimulation

250 ng/mL rhCC16 treatment decreased positive rate to β-galactosidase staining (

0.05)

reduced mRNA and protein levels of P16 and P21(

0.05) and protein levels of p-P53

p-P38 and p-ERK1/2(

0.05).

Conclusion

rhCC16 inhibits CSE-induced cellular senescence of HBECs

which may due to its suppression in P38 MAPK and ERK1/2 signaling pathway.

关键词

Keywords

references

HOGG J C, TIMENS W. The pathology of chronic obstructive pulmonary disease[J]. Annu Rev Pathol, 2009, 4: 435-459. DOI: 10.1146/annurev.pathol.4.110807.092145.

WU C T, LI G H, HUANG C T, et al . Acute exacerbation of a chronic obstructive pulmonary disease prediction system using wearable device data, machine learning, and deep learning: development and cohort study[J ] . JMIR Mhealth Uhealth, 2021, 9(5): e22591. DOI: 10.2196/22591.

WU H, MA H M, WANG L M, et al . Regulation of lung epithelial cell senescence in smoking-induced COPD/emphysema by microR-125a-5p via Sp1 mediation of SIRT1/HIF-1a[J ] . Int J Biol Sci, 2022, 18(2): 661-674. DOI: 10.7150/ijbs.65861.

BARNES P J, BAKER J, DONNELLY L E. Cellular senescence as a mechanism and target in chronic lung diseases[J]. Am J Respir Crit Care Med, 2019, 200(5): 556-564. DOI: 10.1164/rccm.201810-1975TR.

AGHALI A, KOLOKO NGASSIE M L, PABELICK C M, et al . Cellular senescence in aging lungs and diseases[J ] . Cells, 2022, 11(11): 1781. DOI: 10.3390/cells11111781.

CORTOPASSI F, GURUNG P, PINTO-PLATA V. Chronic obstructive pulmonary disease in elderly patients[J]. Clin Geriatr Med, 2017, 33(4): 539-552. DOI: 10.1016/j.cger.2017.06.006.

BROECKAERT F, BERNARD A. Clara cell secretory protein (CC16): characteristics and perspectives as lung peripheral biomarker[J ] . Clin Exp Allergy, 2000, 30(4): 469-475. DOI: 10.1046/j.1365-2222.2000.00760.x.

BOERS J E, AMBERGEN A W, THUNNISSEN F B. Number and proliferation of clara cells in normal human airway epithelium[J]. Am J Respir Crit Care Med, 1999, 159(5 Pt 1): 1585-1591. DOI: 10.1164/ajrccm.159.5.9806044.

GRIBBEN K C, POOLE J A, NELSON A J, et al . Relationships of serum CC16 levels with smoking status and lung function in COPD[J ] . Respir Res, 2022, 23(1): 247. DOI: 10.1186/s12931-022-02158-8.

LAUCHO-CONTRERAS M E, POLVERINO F, ROJAS-QUINTERO J, et al . Club cell protein 16 (Cc16) deficiency increases inflamm-aging in the lungs of mice[J ] . Physiol Rep, 2018, 6(15): e13797. DOI: 10.14814/phy2.13797.

LAUCHO-CONTRERAS M E, POLVERINO F, TESFAIGZI Y, et al . Club cell protein 16 (CC16) augmentation: a p otential disease-modifying approach for chronic obstructive pulmonary disease (COPD)[J ] . Expert Opin Ther Targets, 2016, 20(7): 869-883. DOI: 10.1517/14728222.2016.1139084.

秦轲茹. 重组CC16蛋白质抑制香烟烟雾诱导炎症反应的初步研究[D]. 太原: 山西医科大学, 2022. DOI: 10.27288/d.cnki.gsxyu.2022.001152.

QIN K R. Preliminary study on recombinant CC16 protein inhibiting inflammatory response induced by cigarette smoke[D]. Taiyuan: Shanxi Medical University, 2022.

廖科, 陈亚娟, 赵绿翠, 等. 烟草烟雾提取物通过TGF-β1/Smad2通路诱导RLE-6TN细胞凋亡[J]. 第三军医大学学报, 2015, 37(14): 1412-1416. DOI: 10.16016/j.1000-5404.201411195.

LIAO K, CHEN Y J, ZHAO L C, et al . Cigarette smoke extract induces apoptosis of RLE-6TN cells via TGF-β1/Smad2 pathway[J ] . J Third Mil Med Univ, 2015, 37(14): 1412-1416. DOI: 10.16016/j.1000-5404.201411195.

LOPEZ-OTIN C, BLASCO M A, PARTRIDGE L, et al . The hallmarks of aging[J ] . Cell, 2013, 153(6): 1194-1217. DOI: 10.1016/j.cell.2013.05.039.

MUNOZ-ESPIN D, SERRANO M. Cellular senescence: from physiology to pathology[J]. Nat Rev Mol Cell Biol, 2014, 15(7): 482-496. DOI: 10.1038/nrm3823.

VAN DEURSEN J M. The role of senescent cells in ageing[J]. Nature, 2014, 509(7501): 439-446. DOI: 10.1038/nature13193.

ITAHANA K, CAMPISI J, DIMRI G P. Methods to detect biomarkers of cellular senescence: the senescence-associated beta-galactosidase assay[J]. Methods Mol Biol, 2007, 371: 21-31. DOI: 10.1007/978-1-59745-361-5_3.

TSUJI T, AOSHIBA K, NAGAI A. Alveolar cell senescence in patients with pulmonary emphysema[J]. Am J Respir Crit Care Med, 2006, 174(8): 886-893. DOI: 10.1164/rccm.200509-1374OC.

AMSELLEM V, GARY-BOBO G, MARCOS E, et al . Telomere dysfunction causes sustained inflammation in chronic obstructive pulmonary disease[J ] . Am J Respir Crit Care Med, 2011, 184(12): 1358-1366. DOI: 10.1164/rccm.201105-0802OC.

RUTTEN E P A, GOPAL P, WOUTERS E F M, et al . Various mechanisticpathways representing the aging process are altered in COPD[J ] . Chest, 2016, 149(1): 53-61. DOI: 10.1378/chest.15-0645.

PANG M, LIU H Y, LI T, et al . Recombinant club cell protein 16 (CC16) ameliorates cig arette smoke-induced lung inflammation in a murine disease model of COPD[J ] . Mol Med Rep, 2018, 18(2): 2198-2206. DOI: 10.3892/mmr.2018.9216.

BANERJEE A, KOZIOL-WHITE C, PANETTIERI R. p38 MAPK inhibitors, IKK2 inhibitors, and TNFα inhibitors in COPD[J]. Curr Opin Pharmacol, 2012, 12(3): 287-292. DOI: 10.1016/j.coph.2012.01.016.

HERRANZ N, GALLAGE S, MELLONE M, et al . mTOR regulates MAPKAPK2 translation to control the senescence-associated secretory phenotype[J ] . Nat Cell Biol, 2015, 17(9): 1205-1217. DOI: 10.1038/ncb3225.

NIU L L, FANG Y, YAO X Q, et al . TNFα activates MAPK and Jak-Stat pathways to promote mouse Müller cell proliferation[J ] . Exp Eye Res, 2021, 202: 108353. DOI: 10.1016/j.exer.2020.108353.

ZHANG H H, LIU J L, DANG Q Q, et al . Ribosomal protein RPL5 regulates colon cancer cell proliferation and migration through MAPK/ERK signaling pathway[J ] . BMC Mol Cell Biol, 2022, 23(1): 48. DOI: 10.1186/s12860-022-00448-z.

DEBACQ-CHAINIAUX F, BOILAN E, DEDESSUS LE MOUTIER J, et al . p38(MAPK) in the senescence of human and murine fibroblasts[J ] . A dv Exp Med Biol, 2010, 694: 126-137. DOI: 10.1007/978-1-4419-7002-2_10.

DESCHEÊNES-SIMARD X, GAUMONT-LECLERC M F, BOURDEAU V, et al . Tumor suppressor activity of the ERK/MAPK pathway by promoting selective protein degradation[J ] . Genes Dev, 2013, 27(8): 900-915. DOI: 10.1101/gad.203984.112.

郭民, 李婷, 杨晓雪, 等. 重组人CC16蛋白改善慢性阻塞性肺疾病模型小鼠肺功能及其机制[J]. 中国病理生理杂志, 2023, 39(10): 1789-1795. DOI: 10.3969/j.issn.1000-4718.2023.10.008.

GUO M, LI T, YANG X X, et al . Recombinant human CC16 protein improves lung function in chronic ob-structive pulmonary disease mice and its mechanism[J ] . Chin J Pathophysiol, 2023, 39(10): 1789-1795. DOI: 10.3969/j.issn.1000-4718.2023.10.008.

JOHNSON M D L, YOUNIS U S, MENGHANI S V, et al . CC16 binding to α 4 β 1 integrin protects against Mycoplasma pneumoniae infection[J ] . Am J Respir Crit Care Med, 2021, 203(11): 1410-1418. DOI: 10.1164/rccm.202006-2576OC..

ZHOU Z, ZHOU Q, WU X, et al . VCAM-1 secreted from cancer-associated fibroblasts enhances the growth and invasion of lung cancer cells through AKT and MAPK signaling[J ] . Cancer Lett, 2020, 473: 62-73. DOI: 10.1016/j.canlet.2019.12.039.

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