morphoМорфологические ведомостиMorphological newsletter1812-31712686-8741Private Medical University REAVIZ10.20340/mv-mn.2023.31(3).794morpho-794Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯRESEARCH ARTICLESМОРФОФУНКЦИОНАЛЬНАЯ ОЦЕНКА FAP+ И α-SMA+-КЛЕТОК В РАЗЛИЧНЫЕ СРОКИ ТОКСИЧЕСКОГО ФИБРОЗА ПЕЧЕНИ У КРЫСTHE MORPHOLOGICAL AND FUNCTIONAL ASSESSMENT OF FAP+ AND α-SMA+-CELLS IN DIFFERENT TIMES OF RAT TOXIC LIVER FIBROSISЛебедеваЕлена ИвановнаLebedevaElena I.

кандидат биологических наук, доцент кафедры гистологии, цитологии и эмбриологии

Candidate of Biological Sciences, Associate Professor of the Department of Histology, Cytology and Embryology

lebedeva.ya-elenale2013@yandex.byВитебский государственный медицинский университет, Витебск, Республика БеларусьБеларусьVitebsk State Medical University, Vitebsk, Republic of BelarusBelarus202305082023313МОРФОЛОГИЧЕСКИЕ ВЕДОМОСТИ - MORPHOLOGICAL NEWSLETTER 2023-31-41422Copyright © Лебедева Е.И., 20232023Лебедева Е.И.Lebedeva E.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.morpholetter.com/jour/article/view/794

Углубленное изучение источника фиброгенной популяции клеток применительно к этиологии и стадии фиброза, а также понимание молекулярных механизмов, регулирующих изменение фенотипа печеночных фибробластов, имеют первостепенное значение при разработке фармакологических препаратов. Цель исследования – морфофункциональная оценка активированных портальных фибробластов (FAP+) и жир-накапливающих клеток (α-SMA+) печени в различные сроки токсического фиброза печени крыс. Фиброз и цирроз печени у крыс-самцов Wistar индуцировали раствором тиоацетамида в течение 17 недель. Морфологическое исследование печени проводили на парафиновых срезах, окрашенных гематоксилином и эозином, методом Маллори, иммуногистохимическое исследование производили с помощью поликлональных кроличьих антител к антигену портальных фибробластов FAP и с помощью моноклональных мышиных антител к антигену α-SMA+ клеток. До наступления стадии фиброза печени F3/F4 с 3 по 7 неделю количество FAP+ и α-SMA+-клеток увеличивалось попеременно. На этапах трансформации фиброза в цирроз с 7 по 11 неделю повышение их количества происходило незначительно. На стадии неполного (F5) и до наступления достоверного цирроза (F6) с 11 по 15 неделю количество FAP+ и α-SMA+-клеток было непостоянным и происходило попеременное увеличение и уменьшение их количества. α-SMA+-клетки до начала процесса трансформации фиброза в цирроз (F4/F5) наблюдали в синусоидах и очагах некроза. Затем они выявлялись как в синусоидах, так и в соединительнотканных трабекулах. FAP+-клетки на стадии портального фиброза (F1) локализовались около междольковых сосудов и междольковых желчных протоков портальных зон, а с периода F2/F3 их выявляли в соединительнотканных трабекулах и синусоидах. В количественном отношении на всех стадиях фиброза преобладали α-SMA+-клетки. На основании полученных результатов можно предположить, что FAP+-клетки вносят основной вклад в развитие портального и начального этапа мостовидного фиброза. Их следует рассматривать как одну из популяций миофибробластов при тиоацетамид-индуцированном фиброгенезе печени.

Qualitative study of the source of the fibro-genic cell population in relation to the etiology and stage of fibrosis, as well as an understanding of the molecular mechanisms that regulate changes in the phenotype of hepatic fibroblasts, are of paramount importance in the development of pharmacological drugs. The purpose of the study was a morphological and functional assessment of activated portal fibroblasts (FAP+) and fat-accumulating cells (α-SMA+) of the liver at various stages of toxic liver fibrosis in rats. Liver fibrosis and cirrhosis in male Wistar rats were induced with thioacetamide solution for 17 weeks. Morphological examination of the liver was carried out on paraffin sections stained with hematoxylin and eosin using the Mallory method; immunohistochemical examination was carried out using polyclonal rabbit antibodies to the portal fibroblast antigen FAP and using monoclonal mouse antibodies to the α-SMA+ cell antigen. Before the onset of liver fibrosis stage F3/F4, from weeks 3 to 7, the number of FAP+ and α-SMA+ cells increased alternately. During the stages of transformation of fibrosis into cirrhosis from 7 to 11 weeks, their number increased slightly. At the stage of incomplete (F5) and before the onset of significant cirrhosis (F6) from weeks 11 to 15, the number of FAP+ and α-SMA+ cells were inconsistent and there was an alternating increase and decrease in their number. α-SMA+ cells before the start of the process of transformation of fibrosis into cirrhosis (F4/F5) were observed in sinusoids and foci of necrosis. Then they were detected both in sinusoids and in connective tissue trabeculae. FAP+ cells at the stage of portal fibrosis (F1) were localized near the interlobular vessels and interlobular bile ducts of the portal zones, and from the F2/F3 period they were detected in connective tissue trabeculae and sinusoids. In quantitative terms, α-SMA+ cells predominated at all stages of fibrosis. Based on the results obtained, it can be assumed that FAP+ cells make a major contribution to the development of the portal and initial stages of bridging fibrosis. They should be considered as one of the myofibroblast populations in thioacetamide-induced liver fibrogenesis.

печенькрысыфиброзFAP+ клеткиα-SMA+ клеткиliverratsfibrosisFAP+ cellsα-SMA+ cellsИсследование выполнено при финансовой поддержке Министерства здравоохранения Республики Беларусь в рамках государственной программы научных исследований «Фундаментальные и прикладные науки – медицине», проект № 20190107The study was financially supported by the Ministry of Health of the Republic of Belarus within the framework of the state research program "Fundamental and applied sciences for medicine", Research № 20190107ReferencesBaglieri J, Brenner DA, Kisseleva T. The Role of Fibrosis and Liver-Associated Fibroblasts in the Pathogenesis of Hepatocellular Carcinoma. Int J Mol Sci. 2019;20(7):1723.DOI:10.3390/ijms20071723Baglieri J, Brenner DA, Kisseleva T. The Role of Fibrosis and Liver-Associated Fibroblasts in the Pathogenesis of Hepatocellular Carcinoma. Int J Mol Sci. 2019;20(7):1723.DOI:10.3390/ijms20071723Lay AJ, Zhang HE, McCaughan GW et al. Fibroblast activation protein in liver fibrosis. Front Biosci (Landmark Ed). 2019;24(1):1-17. DOI:10.2741/4706Lay AJ, Zhang HE, McCaughan GW et al. Fibroblast activation protein in liver fibrosis. Front Biosci (Landmark Ed). 2019;24(1):1-17. DOI:10.2741/4706Dhar D, Baglieri J, Kisseleva T et al. Mechanisms of liver fibrosis and its role in liver cancer. Exp Biol Med (Maywood). 2020;245(2):96-108. DOI:10.1177/1535370219898141Dhar D, Baglieri J, Kisseleva T et al. Mechanisms of liver fibrosis and its role in liver cancer. Exp Biol Med (Maywood). 2020;245(2):96-108. DOI:10.1177/1535370219898141Luo N, Li J, Wei Y et al. Hepatic Stellate Cell: A Double-Edged Sword in the Liver. Physiol Res. 2021;70(6):821-829. DOI: 10.33549/physiolres.934755Luo N, Li J, Wei Y et al. Hepatic Stellate Cell: A Double-Edged Sword in the Liver. Physiol Res. 2021;70(6):821-829. DOI: 10.33549/physiolres.934755Sufleţel RT, Melincovici CS, Gheban BA et al. Hepatic stellate cells - from past till present: morphology, human markers, human cell lines, behavior in normal and liver pathology. Rom J Morphol Embryol. 2020;61(3):615-642. DOI:10.47162/RJME.61.3.01Sufleţel RT, Melincovici CS, Gheban BA et al. Hepatic stellate cells - from past till present: morphology, human markers, human cell lines, behavior in normal and liver pathology. Rom J Morphol Embryol. 2020;61(3):615-642. DOI:10.47162/RJME.61.3.01Sørensen MD, Thiele M, Krag A et al. Stage-dependent expression of fibrogenic markers in alcohol-related liver disease. Pathol Res Pract. 2022;231:153798. DOI: 10.1016/j.prp.2022.153798Sørensen MD, Thiele M, Krag A et al. Stage-dependent expression of fibrogenic markers in alcohol-related liver disease. Pathol Res Pract. 2022;231:153798. DOI: 10.1016/j.prp.2022.153798Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev. 2017;121:27-42. DOI: 10.1016/j.addr.2017.05.007Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev. 2017;121:27-42. DOI: 10.1016/j.addr.2017.05.007Sun Y, Liu B, Xie J et al. Aspirin attenuates liver fibrosis by suppressing TGF β1/Smad signaling. Mol Med Rep. 2022;25(5):181. DOI:10.3892/mmr.2022.12697Sun Y, Liu B, Xie J et al. Aspirin attenuates liver fibrosis by suppressing TGF β1/Smad signaling. Mol Med Rep. 2022;25(5):181. DOI:10.3892/mmr.2022.12697Wells RG. Portal Fibroblasts in Biliary Fibrosis. Curr Pathobiol Rep. 2014;2(4):185-190. DOI: 10.1007/s40139-014-0054-yWells RG. Portal Fibroblasts in Biliary Fibrosis. Curr Pathobiol Rep. 2014;2(4):185-190. DOI: 10.1007/s40139-014-0054-yFuji H, Miller G, Nishio T et al. The role of mesothelin signaling in portal fibroblasts in the pathogenesis of cholestatic liver fibrosis. Front Mol Biosci. 2021;8:790032. DOI: 10.3389/fmolb.2021.790032Fuji H, Miller G, Nishio T et al. The role of mesothelin signaling in portal fibroblasts in the pathogenesis of cholestatic liver fibrosis. Front Mol Biosci. 2021;8:790032. DOI: 10.3389/fmolb.2021.790032Yang AT, Kim YO, Yan XZ et al. Fibroblast Activation Protein Activates Macrophages and Promotes Parenchymal Liver Inflammation and Fibrosis. Cell Mol Gastroenterol Hepatol. 2022:S2352-345X(22)00253-3. DOI: 10.1016/j.jcmgh.2022.12.005Yang AT, Kim YO, Yan XZ et al. Fibroblast Activation Protein Activates Macrophages and Promotes Parenchymal Liver Inflammation and Fibrosis. Cell Mol Gastroenterol Hepatol. 2022:S2352-345X(22)00253-3. DOI: 10.1016/j.jcmgh.2022.12.005Banin VV, Belousova TA, Bykov VL i dr. Terminologia Histologica. Mezhdunarodnye terminy po tsitologii i gistologii cheloveka s ofitsial'nym spiskom russkikh ekvivalentov: spravochnoe posobie. M.: GEOTAR-Media, 2009.- 272s. In RussianBanin VV, Belousova TA, Bykov VL i dr. Terminologia Histologica. Mezhdunarodnye terminy po tsitologii i gistologii cheloveka s ofitsial'nym spiskom russkikh ekvivalentov: spravochnoe posobie. M.: GEOTAR-Media, 2009.- 272s. In RussianEverhart JE, Wright EC, Goodman ZD et al. HALT-C Trial Group. Prognostic value of Ishak fibrosis stage: findings from the hepatitis C antiviral long-term treatment against cirrhosis trial // Hepatology. 2010. Vol. 51, N 2. P. 585-594. DOI: 10.1002/hep.23315Everhart JE, Wright EC, Goodman ZD et al. HALT-C Trial Group. Prognostic value of Ishak fibrosis stage: findings from the hepatitis C antiviral long-term treatment against cirrhosis trial // Hepatology. 2010. Vol. 51, N 2. P. 585-594. DOI: 10.1002/hep.23315Kirik OV, Karpenklo MN, Korzhevskiy DE. Teoreticheskie osnovy i prakticheskoe primenenie metodov immunogistokhimii. Rukovodstvo. S-Pb.: SpetsLit, 2014.- 119s. In RussianKirik OV, Karpenklo MN, Korzhevskiy DE. Teoreticheskie osnovy i prakticheskoe primenenie metodov immunogistokhimii. Rukovodstvo. S-Pb.: SpetsLit, 2014.- 119s. In RussianAvery D, Govindaraju P, Jacob M et al. Extracellular matrix directs phenotypic heterogeneity of activated fibroblasts. Matrix Biol. 2018;67:90-106. DOI: 10.1016/j.matbio.2017.12.003Avery D, Govindaraju P, Jacob M et al. Extracellular matrix directs phenotypic heterogeneity of activated fibroblasts. Matrix Biol. 2018;67:90-106. DOI: 10.1016/j.matbio.2017.12.003Younis NS, Ghanim AMH, Elmorsy MA et al. Taurine ameliorates thioacetamide induced liver fibrosis in rats via modulation of toll like receptor 4/nuclear factor kappa B signaling pathway. Sci Rep. 2021 Jun 10;11(1):12296. DOI: 10.1038/s41598-021-91666-6Younis NS, Ghanim AMH, Elmorsy MA et al. Taurine ameliorates thioacetamide induced liver fibrosis in rats via modulation of toll like receptor 4/nuclear factor kappa B signaling pathway. Sci Rep. 2021 Jun 10;11(1):12296. DOI: 10.1038/s41598-021-91666-6Voutilainen SH, Kosola SK, Lohi J et al. Expression of 6 Biomarkers in Liver Grafts After Pediatric Liver Transplantation: Correlations with Histology, Biochemistry, and Outcome. Ann Transplant. 2020;25:e925980. DOI: 10.12659/AOT.925980Voutilainen SH, Kosola SK, Lohi J et al. Expression of 6 Biomarkers in Liver Grafts After Pediatric Liver Transplantation: Correlations with Histology, Biochemistry, and Outcome. Ann Transplant. 2020;25:e925980. DOI: 10.12659/AOT.925980Fitzgerald AA, Weiner LM. The role of fibroblast activation protein in health and malignancy. Cancer Metastasis Rev. 2020;39(3):783-803. DOI: 10.1007/s10555-020-09909-3Fitzgerald AA, Weiner LM. The role of fibroblast activation protein in health and malignancy. Cancer Metastasis Rev. 2020;39(3):783-803. DOI: 10.1007/s10555-020-09909-3Wu Y, Li Z, Xiu AY et al. Carvedilol attenuates carbon tetrachloride-induced liver fibrosis and hepatic sinusoidal capillarization in mice Drug Des Devel Ther. 2019;13:2667-2676. DOI: 10.2147/DDDT.S210797Wu Y, Li Z, Xiu AY et al. Carvedilol attenuates carbon tetrachloride-induced liver fibrosis and hepatic sinusoidal capillarization in mice Drug Des Devel Ther. 2019;13:2667-2676. DOI: 10.2147/DDDT.S210797Lebedeva EI. The Role of CK19-Positive Portal Zone Cells in Thioacetamide Induced Rat Liver Cirrhosis. Cell and Tissue Biology. 2021;15(6):568-576. DOI: 10.1134/S1990519X21060067Lebedeva EI. The Role of CK19-Positive Portal Zone Cells in Thioacetamide Induced Rat Liver Cirrhosis. Cell and Tissue Biology. 2021;15(6):568-576. DOI: 10.1134/S1990519X21060067Lemoinne S, Thabut D, Housset C. Portal myofibroblasts connect angiogenesis and fibrosis in liver. Cell Tissue Res. 2016;365(3):583-589. DOI: 10.1007/s00441-016-2443-5Lemoinne S, Thabut D, Housset C. Portal myofibroblasts connect angiogenesis and fibrosis in liver. Cell Tissue Res. 2016;365(3):583-589. DOI: 10.1007/s00441-016-2443-5Dendl K, Koerber SA, Kratochwil C et al. FAP and FAPI-PET/CT in Malignant and Non-Malignant Diseases: A Perfect Symbiosis? Cancers (Basel). 2021;13(19):4946. DOI: 10.3390/cancers13194946Dendl K, Koerber SA, Kratochwil C et al. FAP and FAPI-PET/CT in Malignant and Non-Malignant Diseases: A Perfect Symbiosis? Cancers (Basel). 2021;13(19):4946. DOI: 10.3390/cancers13194946Fitzgerald AA, Weiner LM. The role of fibroblast activation protein in health and malignancy. Cancer Metastasis Rev. 2020;39(3):783-803. DOI: 10.1007/s10555-020-09909-3Fitzgerald AA, Weiner LM. The role of fibroblast activation protein in health and malignancy. Cancer Metastasis Rev. 2020;39(3):783-803. DOI: 10.1007/s10555-020-09909-3Hamson EJ, Keane FM, Tholen S et al. Understanding fibroblast activation protein (FAP): substrates, activities, expression and targeting for cancer therapy. Proteomics Clin Appl. 2014;8(5-6):454-463. DOI: 10.1002/prca.201300095Hamson EJ, Keane FM, Tholen S et al. Understanding fibroblast activation protein (FAP): substrates, activities, expression and targeting for cancer therapy. Proteomics Clin Appl. 2014;8(5-6):454-463. DOI: 10.1002/prca.201300095

The authors declare that there are no conflicts of interest present.