Effect of p38 MAPK Inhibition on Apoptosis Marker Expression in the Process of Peritoneal Adhesion Formation

Irina A. Shurygina, Nataliya I. Aushinova, Michael G. Shurygin

International Journal of Biomedicine. 2018;8(4):342-346.   
DOI: 10.21103/Article8(4)_OA15
Originally published December 15, 2018  


Background: Apoptosis, one of the most important mechanisms for maintaining homeostasis, is carried out under both physiological and pathological conditions. The aim of our study was to investigate the expression of markers of apoptosis through caspase-dependent and caspase-independent pathways during the reparative regeneration after serosal injuries of the peritoneum in the context of the prolonged p38 MAPK inhibition.
Methods: Peritoneal adhesions in the animal models were induced by a method developed by the authors that included opening the serous-muscular layer of the caecum with a 1cm cut followed by closing the wound with a Schmieden suture and scarifying a 1.5×1.5 cm area of the parietal peritoneum of the right lateral channel. Experiments were carried out on male 9-month-old Wistar rats. When closing the wound, control animals were intraperitoneally administered 3 mL of 0.9% sodium chloride solution (n=40) while experimental group rats were administered slow-release drug Seroguard® (Pharmasyntez JSC) (n=40). Animals were sacrificed within the period of 2 hours to 30 days post surgery. Expression of apoptosis markers was studied by immunohistochemical (Bcl-2, Bcl-x) and immunofluorescent (PARP-1) staining.
Results: It is interesting that, in cases of the natural regeneration of the peritoneal injury, expression of anti-apoptosis markers at the injury site came in two waves: it was the most pronounced on days 1–3 post surgery while the second peak of activity was observed on day 14. Within this time window, granulation tissue was actively growing and mature connective-tissue vascularized adhesions were being formed. By the end of the observation period (day 30), expression of anti-apoptosis proteins at the injury site became extremely low and a significant reduction in the amount of connective tissue cells was observed. It was found that a prolonged inhibition of the p38 activity resulted in a moderate increase in Bcl-2 expression on days 3–7, and a decrease in the activity on day 14 was followed by another increase in expression by day 30. The Bcl-xl expression was observed 12 hours to 3 days post surgery and then it went down to the minimum. Positive PARP-1 staining observed on days 3 to 30, which reached its maximum on day 14, was also typical of the experimental group.
Conclusion: The performed study demonstrated that a prolonged p38 MAPK inhibition in the adhesion formation models results in the activation of fibroblast apoptosis at the reparation site, which, in the authors’ opinion, predetermines a significant decrease in the adhesion formation in the experimental group.

adhesions • peritoneal cavity • p38 MAPK • apoptosis • Seroguard • Bcl-2 • Bcl-xl • PARP-1

1. Karp G. Cell and molecular biology: Concepts and experiments. 6th ed. John Wiley&Sons, Hoboken, NY; 2010.
2. Gordeeva AV, Labas YA, Zvyagilskaya RA. Apoptosis in unicellular organisms: mechanisms and evolution. Biochemistry (Mosc). 2004;69(10):1055-66. PubMed
3. Popgeorgiev N, Jabbour L, Gillet G. Subcellular Localization and Dynamics of the Bcl-2 Family of Proteins. Front Cell Dev Biol. 2018;6:13. doi: 10.3389/fcell.2018.00013. PubMed 
4. Kale J, Osterlund K, Andrews DW. BCL-2 family proteins: changing partners in the dance towards death. Cell Death Differ. 2018;25(1):65-80. doi: 10.1038/cdd.2017.186.  PubMed
5. Chao DT, Korsmeyer SJ. BCL-2 family: regulators of cell death. Annu Rev Immunol. 1998;16:395-419. PubMed
6. Siddiqui WA, Ahad A, Ahsan H. The mystery of BCL2 family: Bcl-2 proteins and apoptosis: an update. Arch Toxicol. 2015;89(3):289-317. doi: 10.1007/s00204-014-1448-7.  PubMed
7. Yu SW, Andrabi SA, Wang H, Kim NS, Poirier GG, Dawson TM, Dawson VL. Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. Proc Natl Acad Sci U S A. 2006;103(48):18314-9.  PubMed
8. Ling XX, Liu JX, Zhou L, DU YJ, Chen SQ, Chen JL et al. Poly(ADP-ribosyl)ation of Apoptosis Antagonizing Transcription Factor Involved in Hydroquinone-Induced DNA Damage Response. Biomed Environ Sci. 2016;29(1):80-4. doi: 10.3967/bes2016.008. PubMed
9. Desmoulière A, Redard M, Darby I, Gabbiani G. Apoptosis mediates the decrease in cellularity during the transition between granulation tissue and scar. Am J Pathol. 1995; 146(1):56-66.  PubMed
10. Hinz B. Formation and function of the myofibroblast during tissue repair. J Invest Dermatol. 2007;127(3):526-37. PubMed
11. Ayushinova NI, Lepekhova SA, Shurygina IA, Roj TA, Shurygin MG, Zaritskaja LV, Gol'dberg OA. Method for simulating abdominal commissures. Patent RU 2467401, 27.07.2011.
12. Ayushinova NI, Shurygina IA, Shurygin MG, Balykina AV, Malgataeva ER, Popova AD, Yankelevich SA. Experimental model for developing of ways to prevent adhesions in the abdominal cavity. Siberian Med J. 2012;109(2):51-53. Google Scholar
13. Shurygina IA, Shurygin MG, Rodionova LV, Aushinova NI. Interleukin expression in the area damaged by the development of abdominal cavity adhesions. International Journal of Biomedicine. 2017;7(4):293-297. doi: 10.21103/Article7(4)_OA4.  Google Scholar
14. Shurygin MG, Shurygina IA. Compounds, pharmaceutical compositions and a method for the prophylaxis and treatment of the adhesion process. Patent WO2012156938, 20.03.2014.
15. diZerega GS. Peritoneum, peritoneal healing and adhesion formation. In: diZerega GS, editor. Peritoneal surgery. Berlin-Heidelberg-New York: Springer; 2006:3-38.
16. Shurygin MG, Shurygina IA, Dremina NN, Kanya OV. Endogenous progenitors as the source of cell material for ischemic damage repair in experimental myocardial infarction under conditions of changed concentration of vascular endothelial growth factor. Bull Exp Biol Med. 2015; 158(4):528-31. doi: 10.1007/s10517-015-2801-4. PubMed
17. Binnebösel M, Klinge U, Rosch R, Junge K, Lynen-Jansen P, Schumpelick V. Morphology, quality, and composition in mature human peritoneal adhesions. Langenbecks Arch Surg. 2008;393(1):59-66.  PubMed
18. Saed GM, Jiang Z, Fletcher NM, Diamond MP. Modulation of the BCL-2/BAX ratio by interferon-gamma and hypoxia in human peritoneal and adhesion fibroblasts. Fertil Steril. 2008;90(5):1925-30.  PubMed
19. Shurygina IA, Shurygin MG, Ayushinova NI. Expression of apoptosis markers in adhesions in the abdominal cavity under the experimental conditions. Vestnik Rossiiskoi Akademii Meditsinskikh Nauk. 2014;(5-6):29-33.

Download Article
Received September 15, 2018.
Accepted October 22, 2018.
©2018 International Medical Research and Development Corporation.