Ukr.Biochem.J. 2022; Volume 94, Issue 4, Jul-Aug, pp. 36-46
doi: https://doi.org/10.15407/ubj94.04.036
Localization and level of proapoptotic protein regulators in a rat lung tissue during development of acute experimental bronchopneumonia
D. S. Ziablitsev1*, A. O. Tykhomyrov2, O. O. Dyadyk3,
S. V. Kolesnikova1, S. V. Ziablitsev1
1Bogomolets National Medical University, Kyiv, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Shupyk National Healthcare University of Ukraine, Kyiv;
*e-mail: denis898@ukr.net
Received: 20 July 2022; Revised: 13 September 2022;
Accepted: 04 November 2022; Available on-line: 14 November 2022
Apoptosis plays an important role in the development of acute inflammatory lung injury (AILI) and its consequences, which can be realized in different cells with distinct intensity and rate. The aim of this study was to determine the distribution and expression intensity of apoptosis markers in the lungs of rats in the AILI model with endotracheal introduction of capron thread and LPS. Immunoblotting and immunohistochemical studies were performed using monoclonal antibodies against Bax and caspase-3 proteins. It was shown that Bax level increased significantly with the peak on the 7th day. The second peak of Bax 40 dimeric form was noted on the 21st day. The level of both pro-caspase-3 and active caspase-3 was also dramatically increased with a maximum on the 5th day and the second peak of active caspase-3 content was observed on the 21st day. These changes reflected the activation of apoptosis in key trigger periods of AILI during the development of exudative hemorrhagic pneumonia and subsequent fibrotic remodeling of the lungs.
Keywords: AILI, apoptosis, Bax, caspase-3, lipopolysaccharide
References:
- World Health Organization. Coronavirus disease (COVID-19) pandemic. Available et https://www.who.int/emergencies/diseases/novel-coronavirus-2019 (accessed, July, 2022).
- Cai A, McClafferty B, Benson J, Ramgobin D, Kalayanamitra R, Shahid Z, Groff A, Aggarwal CS, Patel R, Polimera H, Vunnam R, Golamari R, Sahu N, Bhatt D, Jain R. COVID-19: Catastrophic Cause of Acute Lung Injury. S D Med. 2020;73(6):252-260. PubMed
- Suster S, Moran AC. Biopsy interpretation of the lung. 1st ed. Lippincott Williams & Wilkins, Wolters Kluwer; 2013. 417 p.
- Rybakova MG, Karev VE, Kuznetsova IA. Anatomical pathology of novel coronavirus (COVID-19) infection. First impressions. Arkh Patol. 2020;82(5):5-15. (In Russian). PubMed, CrossRef
- Ackermann M, Verleden SE, Kuehnel M, Haverich A, Welte T, Laenger F, Vanstapel A, Werlein C, Stark H, Tzankov A, Li WW, Li VW, Mentzer SJ, Jonigk D. Pulmonary Vascular Endothelialitis, Thrombosis, and Angiogenesis in Covid-19. N Engl J Med. 2020;383(2):120-128. PubMed, PubMedCentral, CrossRef
- Lu Q, Harrington EO, Rounds S. Apoptosis and lung injury. Keio J Med. 2005;54(4):184-189. PubMed, CrossRef
- Chopra M, Reuben JS, Sharma AC. Acute lung injury:apoptosis and signaling mechanisms. Exp Biol Med (Maywood). 2009;234(4):361-371. PubMed, CrossRef
- N’Guessan PD, Schmeck B, Ayim A, Hocke AC, Brell B, Hammerschmidt S, Rosseau S, Suttorp N, Hippenstiel S. Streptococcus pneumoniae R6x induced p38 MAPK and JNK-mediated caspase-dependent apoptosis in human endothelial cells. Thromb Haemost. 2005;94(2):295-303. PubMed, CrossRef
- Neff TA, Guo RF, Neff SB, Sarma JV, Speyer CL, Gao H, Bernacki KD, Huber-Lang M, McGuire S, Hoesel LM, Riedemann NC, Beck-Schimmer B, Zetoune FS, Ward PA. Relationship of acute lung inflammatory injury to Fas/FasL system. Am J Pathol. 2005;166(3):685-694. PubMed, PubMedCentral, CrossRef
- Yamasaki M, Kang HR, Homer RJ, Chapoval SP, Cho SJ, Lee BJ, Elias JA, Lee CG. P21 regulates TGF-beta1-induced pulmonary responses via a TNF-alpha-signaling pathway. Am J Respir Cell Mol Biol. 2008;38(3):346-353. PubMed, PubMedCentral, CrossRef
- Zyablitsev SV, Penskyy PYu, Litvinets ML, Kovalova AV, Salamaha AA. Dynamics of morphological manifestations of the experimental acute aspiration bronchopneumonia development. Morphologia. 2021;15(1):48-59. CrossRef
- Stoscheck CM. Quantitation of protein. Methods Enzymol. 1990;182:50-68. PubMed, CrossRef
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-685. PubMed, CrossRef
- Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA. 1979;76(9):4350-4354. PubMed, PubMedCentral, CrossRef
- Kuwano K, Hagimoto N, Maeyama T, Fujita M, Yoshimi M, Inoshima I, Nakashima N, Hamada N, Watanabe K, Hara N. Mitochondria-mediated apoptosis of lung epithelial cells in idiopathic interstitial pneumonias. Lab Invest. 2002;82(12):1695-1706. PubMed, CrossRef
- Li T , Liu Y, Li G, Wang X, Zeng Z, Cai S, Li F, Chen Z. Polydatin attenuates ipopolysaccharide-induced acute lung injury in rats. Int J Clin Exp Pathol. 2014;7(12):8401-8410. PubMed, PubMedCentral, CrossRef
- Takahara M, Aoyama-Ishikawa M, Shuno K, Yamauhi C, Miyoshi M, Maeshige N, Usami M, Yamada T, Osako T, Nakao A, Kotani J. Role of endogenous IL-18 in the lung during endotoxin-induced systemic inflammation. Acute Med Surg. 2013;1(1):23-30. PubMed, PubMedCentral, CrossRef
- Li X, Yan YH, Feng DY. Apoptosis and caspase-3 in the model of rat silicosis. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2005;30(4):441-443. PubMed
- Seitz DH, Perl M, Mangold S, Neddermann A, Braumüller ST, Zhou S, Bachem MG, Huber-Lang MS, Knöferl MW. Pulmonary contusion induces alveolar type 2 epithelial cell apoptosis: role of alveolar macrophages and neutrophils. Shock. 2008;30(5):537-544. PubMed, CrossRef
- Robb CT, Regan KH, Dorward DA, Rossi AG. Key mechanisms governing resolution of lung inflammation. Semin Immunopathol. 2016;38(4):425-448.
PubMed, PubMedCentral, CrossRef - Belchamber KBR, Hughes MJ, Spittle DA, Walker EM, Sapey E. New Pharmacological Tools to Target Leukocyte Trafficking in Lung Disease. Front Immunol. 2021;12:704173. PubMed, PubMedCentral, CrossRef
- Nuovo GJ, Magro C, Shaffer T, Awad H, Suster D, Mikhail S, He B, Michaille JJ, Liechty B, Tili E. Endothelial cell damage is the central part of COVID-19 and a mouse model induced by injection of the S1 subunit of the spike protein. Ann Diagn Pathol. 2021;51:151682. PubMed, PubMedCentral, CrossRef
