ASPECTS OF IMMUNOLOGICAL STATUS AND CARBON METABOLISM IN PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE IN CORRELATION WITH ITS FIBROUS TRANSFORMATION
About the author:
Stepanov Y. M., Didenko V. I., Konenko I. S., Tatarchuk O. M., Petishko O. P.
CLINICAL AND EXPERIMENTAL MEDICINE
Type of article:
Today, non-alcoholic fatty liver disease (NAFLD) is becoming more widespread worldwide. NAFLD is diagnosed in almost half of the adult population, 25-30% of whom are at risk to progress to non-alcoholic steatohepatitis. Among the most important factors influencing the prognosis of NAFLD, tactics and effectiveness of treatment is the progression rate of fibrotic liver transformation. Purpose of the investigation: to determine the special features of immunological status changes in patients with NAFLD depending on their stage of fibrosis. Object and methods of investigation. The research involved 101 patients with NAFLD who were undergoing treatment at the Department of Liver and Pancreatic Diseases of «The Institute of Gastroenterology of NAMSU (National Academy of Medical Sciences of Ukraine)». Among the surveyed there were 32 men (31.7%) and 69 women (68.3%), the average age was (48.4±2.9) years. Shear-wave elastography was performed on a Soneus P7 apparatus (Ukraine, Switzerland). Then from these measurements the average values that characterized the liver stiffness in kilopascals (kPa) were determined. Ferraioli G threshold values were used to assess the fibrosis stage. In addition, the same apparatus was used to measure the ultrasound attenuation coefficient in dB/cm – a method of quantitative assessment of the liver steatosis degree. The subpopulation composition of lymphocytes was determined using the monoclonal antibodies of the firm «Sorbent TM» to molecules CD3, CD19, CD4, CD8, CD16. The mononuclear cells were isolated from the peripheral venous blood of patients in a density gradient of 1,077 g/cm. The immune status assessment was performed according to the guidelenes of R.V. Petrov (1992). Circulating immune complexes (CICs) were determined by the method of V. Haskov (1977). During the investigation, sets of reagents of the company «Vector-BEST» were used to measure the amount of TNF-α, IL-6, IL-10 and those of the company «DRG» Germany – to measure insulin in the blood serum. The enzyme-linked immunosorbent assay was performed using a «Stat Fax 303 Plus» analyzer (USA). HOMA-IR = fasting plasma glucose (mmol/l) times fasting serum insulin (mU/ml) devided by 22.5. Research results. While analyzing the immunological indicators in patients with NAFLD it was found that the level of lymphocytes was increased in 38.6% of cases. In 61.4% of patients a significant decrease in the T-helper subpopulation was observed. It should be noted that the median of relative content in CD4+ lymphocytes decreased significantly by 1.3 times (p <0.05) as compared to the control group. In 59.4% of patients with NAFLD, the CIC level was significantly increased (p <0.05) as compared to the control level. According to SWE results, a moderate fibrosis was diagnosed in 52 (51.5%) patients with NAFLD, while 13 (12.9%) patients were diagnosed with marked fibrosis. The highest median values of insulin were observed in patients with severe fibrosis, as evidenced by the correlation between this index and liver stiffness according to SWE data (r = 0.42, p <0.05). In addition, a significant increase in glucose levels of 1.5 times (p <0.05) and 1.3 times (p <0.05) has been observed in NAFLD patients with marked and moderate fibrosis respectively, as compared to the control group levels. The HOMA-IR values in patients with NAFLD were above 3.0 and were significantly different from those in the control group. These changes were more pronounced in patients with marked fibrosis (p <0.05). In accordance with the results of correlation analysis, patients with NAFLD were found to have significant correlations between cellular and humoral immunity indicators, the cytokine level, carbohydrate metabolism and structural changes in the liver. In particular, the detected correlation between the HOMA-IR index values and the attenuation coefficient of the ultrasound (r = 0.56, p <0.01) has proved that NAFLD progression is accompanied by insulin resistance. At the same time, the development of fibrotic transformation in the liver occurs on the background of marked inflammation, as evidenced by the established positive correlations between the liver stiffness index according to SWE and the CIC level (r = 0.56, p <0.01), TNF-α content (r = 0.55, p <0.05) and the ratio value of proinflammatory and anti-inflammatory cytokines (r = 0.50, p <0.05). Conclusions. The progression of liver fibrosis is accompanied by a lack of cellular immunity and an increase in the level of pro-inflammatory cytokines. A significant imbalance between pro-inflammatory and anti-inflammatory cytokines (TNF-α / IL-10) has been detected in patients with marked liver fibrosis. Significant fibrotic liver changes in patients with NAFLD occur on the background of profound disorders in the carbohydrate metabolism.
nonalcoholic fatty liver disease, shear wave elastography, lymphocytes, insulin resistance
- Younossi ZM, Marchesini G, Pinto-Cortez H, Petta S. Epidemiology of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis: implications for liver transplantation. Transplantation. 2019 Jan;103(1):22-7. DOI: 10.1097/TP.0000000000002484
- Sanyal AJ. Past, present and future perspectives in nonalcoholic fatty liver disease. Nat Rev Gastroenterol Hepatol. 2019 Jun;16(6):377-86. DOI: 10.1038/s41575-019-0144-8. Review. PubMed PMID: 31024089.
- Wong VW, Vergniol J, Wong GL, Foucher J, Chan HL, Le Bail B, et al. Diagnosis of fibrosis and cirrhosis using liver stiffness measurement in nonalcoholic fatty liver disease. Hepatology. 2010 Feb;51(2):454-62. DOI: 10.1002/hep.23312. PubMed PMID: 20101745.
- Tatarchuk OM, Didenko VI, Melanich SL, Kudrjavceva VJe. Imunologichna reaktyvnist’ u hvoryh na hronichni dyfuzni zahvorjuvannja pechinky. Gastroenterologija. 2018;4(52):222-6. DOI: https://doi.org/10.22141/23082097.52.4.2018.154142 [in Ukrainian].
- Sljadnev SA. Medyatorу mezhkletochnуh vzaymodeystvyj pry nealkogol’noj zhyrovoj bolezny pecheny. Vestnyk molodogo uchenogo. 2015;3(10):3-8. [in Russiаn].
- Ostanyn AA, Starostyna NM, Meledyna YV, Shypunov MV, Leplyna OJu, Shevela EJa, y dr. Mul’typleksnуj analyz 26 cytokynov, sekretyruemуh kletkamy krovy bol’nуh cyrrozom pecheny. Medycynskaja ymmunologyja. 2015;6(17):539-52. Dostupno: https://doi. org/10.15789/1563- 0625-2015-6-539-552 [in Russiаn].
- Bellot P, Frances R, Such J. Pathological bacterial translocation in cirrhosis: pathophysiology, diagnosis and clinical implications. Liver Int. 2013;1(33):31-9.
- Leibowitz A, Rehman A, Paradis P, Schiffrin EL. Role of T regulatory lymphocytes in the pathogenesis of high-fructose diet-induced metabolic syndrome. Hypertension. 2013;61:1316-21.
- Mohsen Maher, Tarek Yossef, Hesham Darwesh, Ahmed El Saady, Amal I. Sabry, Waled A Hamed, et al. Role of Interleukin 6 as a Predictor of Hepatic encephalopathy in Critically Ill Cirrhotic Patients. Life Sci J. 2013;10(12s):987-91.
- Aldasheva ZhA. Nekotorуe cytokynу v dyagnostyke nealkogol’nogo steatoza pecheny. Cytokynу y vospalenye. 2013;3(12):95-9. [in Russiаn].
- Shadrin OG, Chernega NF, Marushko RV, Brjuzgina TS. Stan zhyrno-kyslotnogo spektru krovi u ditej iz zahvorjuvannjamy pechinky ta jogo zv’jazok z pokaznykamy cytokinovogo statusu. Visnyk problem biologii’ і medycyny. 2015;1(117):193-8. [in Ukraіnian].
- Oh H, Jun DW, Saeed WK, Nguyen MH. Non-alcoholic fatty liver diseases: update on the challenge of diagnosis and treatment. Clin Mol Hepatol. 2016 Sep;22(3):327-35. PubMed PMID: 27729634; PubMed Central PMCID: PMC5066376.
- Wieckowska A, Papouchado BG, Li Z, Lopez R, Zein NN, Feldstein AE. Increased hepatic and circulating interleukin-6 levels in human nonalcoholic steatohepatitis. Am J Gastroenterol. 2008 Jun;103(6):1372-9. DOI: 10.1111/j.1572-0241.2007.01774.x. Epub 2008 May 28. PubMed PMID: 18510618.
- Carter-Kent C, Zein NN, Feldstein AE. Cytokines in the pathogenesis of fatty liver and disease progression to steatohepatitis: implications for treatment. Am. J. Gastroenterol. 2008;103(4):1036-42. DOI: 10.1111/j.1572-0241.2007.01709.x 1
- Glushhenko SV. Optymizacija diagnostyky ta likuvannja nealkogol’nogo steatogepatytu u hvoryh na cukrovyj diabet 2 typu shljahom vyvchennja markeriv mitohondrial’noi’ dysfunkcii’ [dysertatsiya]. Harkiv: 2015. 18 s. [in Ukraіnian].
- Cukanov VV, Kasparov ЕV, Tonkyh JuL, Vasjutyn AV. Novуe aspektу nealkogol’noj zhyrovoj bolezny pecheny. Rossyjskyj zhurnal gastroеnterologyy, gepatologyy, koloproktologyy. 2015;2(25):28-40. [in Russiаn].
Publication of the article:
«Bulletin of problems biology and medicine» Issue 3 (152), 2019 year, 196-200 pages, index UDK 616.36 – 008