Collagens of the third and fourth types in various forms of alcoholic liver disease

The aim of the study was to determine the clinical and diagnostic significance of collagens of the third type (Col3) and the fourth type (Col4) in various forms of alcoholic liver disease (ALD). Materials and methods. 98 patients with ALD were examined: 13 (13.3%) with liver steatosis (LS), 15 (15.3%) with steatohepatitis (SH), 56 (57.1%) with liver cirrhosis (LC), 14 (14.3%) with severe alcoholic hepatitis against the background of liver cirrhosis (SAH-LC). Among the examined there were 59 (60.2%) men and 39 (39.80%) women, the average age was 53.48±11.45 years. The content of fibrogenesis proteins in the blood serum - type III collagen (Col3) and type IV collagen (Col4) was determined by enzyme immunoassay (test systems “Kit For Collagen Type III (Col3)” and “Kit For Collagen Type IV (Col4)”, “Cloud-Clone Corp”, USA), a marker of hepatocyte apoptosis - fragments of cytokeratin-18 (FCK-18) (“Biotech” test system, Sweden), cytokines - IL-1β, IL-4, IL-6, IL- 8, TNF-α (“Vector-Best”, Russia). Results. The level of Col3 in healthy individuals was 6.12±0.73 ng/ml and Col4-9.45±0.32 ng/ml. In all forms of ALD, the content of both types of collagen exceeded that in healthy individuals: Col3 in LS was 6.52±0.94 ng/ml (p<0.05), in SH it was 12.50±3.18 ng/ml (p<0.05), in LC - 20.96±4.93 ng/ml (p<0.05), in SAH-LC - 26.90±3.87 ng/ml (p<0.05); Col4-10.14±0.66 ng/ml, 13.87±1.15 ng/ml, 79.56±33.10 ng/ml and 107.12±39.09 ng/ml, respectively. Col3 positively correlated with ALT (r=0.27, p=0.02) and alkaline phosphatase (r=0.28, p=0.04). Col4 correlated with bilirubin (r=0.74), AST (r=0.65), glutamyl transpeptidase (r=0.58), cytokeratin-18 (r=0.56), sedimentation rate of erythrocytes (r=0.61), C-reactive protein (r=0.59), IL -6 (r= 0.51) and leukocytes (r=0.45) (all p<0.001). Conclusion. Col4 demonstrated greater diagnostic and clinical significance in alcoholic liver disease compared to Col3. The level of Col4 increased by an average of 8 times with the progression of the disease from liver steatosis to cirrhosis, and Col3 - only by 3 times. There were more diverse and stronger associations between Col4 and markers of hepatocellular damage and inflammation than between Col3 and these indicators. The maximum rise in the levels of both types of collagen found in SAH-LC confirmed the important role of this form of liver disease in the development of fibrosis, and hence in the further decompensation of liver cirrhosis.

collagen 3 type, collagen 4 type, fibrosis, alcoholic liver disease, apoptosis, cytokines

1. Asrani S.K., Devarbhavi H., Eaton J., Kamath P. S. Burden of liver diseases in the world. J Hepatol. 2019;70:151-71. doi: 10.1016/j.jhep.2018.09.014.

2. Griswold M. G., Fullman N., Hawley C., Arian N., Zimsen S. R.M., Tymeson H. D. et al. Alcohol use and burden for 195 countries and territories, 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet. 2018;392:1015-1035. doi: 10.1016/S0140-6736(18)31310-2.

3. Karsdal M. A., Daniels S. J., Nielsen S. H., Bager C., Rasmussen D. G.K., Loomba R. et al. Collagen biology and non-invasive biomarkers of liver fibrosis. Liver International. 2020;40:736-750. doi: 10.1111/liv.14390.

4. Karsdal M. A., Hjuler S. T., Luo Y., Rasmussen D. G.K, Nielsen M. J., Nielsen S. H. Assessment of liver fibrosis progression and regression by a serological collagen turnover profile. Am J Physiol Gastrointest Liver Physiol. 2019; 316: G25-G31. doi: 10.1152/ajpgi.00158.2018.

5. Delgado M.E., Cárdenas B. I., Farran N., Fernandez M. Metabolic Reprogramming of Liver Fibrosis. Cells. 2021; 10 (3604). doi: 10.3390/ cells10123604.

6. Friedman S. L., Pinzani M. Hepatic fibrosis 2022: Unmet needs and a blueprint for the future. Hepatology. 2022; 75(2):473-488. doi: 10.1002/hep.32285.

7. Gressner A.M., Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-β as major players and therapeutic targets. J. Cell. Mol. Med. 2006; 10 (1): 76-99. doi: 10.1111/j.1582-4934.2006.tb00292.x.

8. Vinnitskaya E.V., Drozdov V. N., Yunusova Yu.M., Varvanina G. G., Shaposhnikova N. A. Petrakov A. V. et al. Diagnostic significance of serum markers of fibrosis in chronic liver diseases. Therapeutic archive. 2013; 85(2):27-31.@@ Винницкая Е. В., Дроздов В. Н., Юнусова Ю. М., Варванина Г. Г., Шапошникова Н. А. Петраков А.В и др. Диагностическая значимость сывороточных маркеров фиброза при хронических болезнях печени. Терапевтический архив. 2013; 85(2):27-31.

9. Ivanov A.S., Tarasenko E. V., Garmasch I. V., et al. Markers of endothelial dysfunction, cytokine status, collagen COL1A1_1 gene on the development of liver fibrosis in in alcohol abusers. Patologicheskaya Fiziologiya i Eksperimental`naya terapiya. (Pathological Physiology and Experimental Therapy, Russian Journal). 2019; 63(3): 55-63 (in Russ.) doi: 10 25557/0031-2991 2019 03 55-63.@@ Иванов А. С., Тарасенко Е. В., Гармаш И. В., Мяндина Г. И., Аришева О. С., Желудова Е. М. и др. Влияние маркеров эндотелиальной дисфункции, цитокинового статуса, гена коллагена COL1A1_1 на развитие фиброза печени у пациентов, злоупотребляющих алкоголем. Патологическая физиология и экспериментальная терапия. 2019; 63(3): 55-63. DOI: 10.25557/0031-2991.2019.03.55-63.

10. Madsen B. S., Thiele M., Detlefsen S., Kjærgaard M., Møller L. S., Trebicka J. et al. PRO-C3 and ADAPT algorithm accurately identify patients with advanced fibrosis due to alcohol-related liver disease. Alimentary pharmacology and therapeutics. 2021 54(5): 699-708. doi: 10.1111/apt.16513.

11. Nielsen M.J., Kazankov K., Leeming D. J., Karsdal M. A., Krag A., Barrera F. et al. Markers of Collagen Remodeling Detect Clinically Significant Fibrosis in Chronic Hepatitis C Patients. PLoS ONE. 2015; 10 (9): e0137302. doi: 10.1371/journal.pone.0137302.

12. Daniels S.J., Leeming D. J., Eslam M., Hashem A. M., Nielsen M. J. Krag A. et al. ADAPT: an algorithm incorporating PRO-C3 accurately identifies patients with NAFLD and advanced fibrosis. Hepatology. 2019;69:1075-1086. doi: 10.1002/hep.30163.

13. Luo Y.I., Oseini A., Gagnon R., Charles E. D., Sidik K., Vincent R. et al. An evaluation of the collagen fragments related to fibrogenesis and fibrolysis in nonalcoholic steatohepatitis. Sci Rep. 2018;8:12414. doi: 10.1038/s41598-018-30457-y.

14. Boyle M., Tiniakos D., Schattenberg J. M., Ratziu V., Bugianessi E. Petta S. et al. Performance of the PRO-C3 collagen neo-epitope biomarker in non-alcoholic fatty liver disease. JHEP Reports. 2019;1(3):188-198. doi: 10.1016/j.jhepr.2019.06.004.

15. Bril F., Leeming D. J., Karsdal M. A., Kalavalapalli S., Barb D., Lai J. et al. Use of Plasma Fragments of Propeptides of Type III, V, and VI Procollagen for the Detection of Liver Fibrosis in Type 2 Diabetes. Diabetes Care. 2019;42:1348-1351 doi: 10.2337/dc18-2578.

16. Thiele M., Johansen S., Gudmann N. S., Madsen B., Kjærgaard M., Nielsen M. J. et al. Progressive alcohol-related liver fibrosis is characterised by imbalanced collagen formation and degradation. Alimentary pharmacology and therapeutics. 2021; 54 (8):1070-1080. doi: 10.1111/apt.16567.

17. Kerbert A.J.C., Gupta S., Alabsawy E., Dobler I., Lønsmann I., Hall A. Biomarkers of extracellular matrix formation are associated with acute-on-chronic liver failure. JHEP Reports. 2021. 3(6):100355. doi: 10.1016/j.jhepr.2021.100355.

18. Praktiknjo M., Lehmann J., Nielsen M. J., Schierwagen R., Uschner F. E., Meyer C et al. Acute decompensation boosts hepatic collagen type III deposition and deteriorates experimental and human cirrhosis. Hepatology communications. 2018; 2 (2): 211-222. doi: 10.1002/hep4.1135.

19. Hirayama C., Suzuki H., Takada A., Fujisawa K., Tanikawa K., Igarashi S. Serum type IV collagen in various liver diseases in comparison with serum 7S collagen, laminin, and type III procollagen peptide. J Gastroenterol. 1996; 31: 242-248. doi: 10.1007/BF02389524.

20. Roehlen N., Crouchet E., Baumert T. F. Liver Fibrosis: Mechanistic Concepts and Therapeutic Perspectives. Cells. 2020; 9 (4), 875. doi: 10.3390/cells9040875.