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The Effects of Betaine on the Nuclear Fractal Dimension, Chromatin Texture, and Proliferative Activity in Hepatocytes in Mouse Model of Nonalcoholic Fatty Liver Disease

Published online by Cambridge University Press:  19 January 2018

Milena Vesković
Affiliation:
Institute of Pathophysiology, Ljubodrag Buba Mihailović, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Milica Labudović-Borović
Affiliation:
Institute of Histology and Embryology, Aleksandar Đ. Kostić, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Ivan Zaletel
Affiliation:
Institute of Histology and Embryology, Aleksandar Đ. Kostić, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Jelena Rakočević
Affiliation:
Institute of Histology and Embryology, Aleksandar Đ. Kostić, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Dušan Mladenović
Affiliation:
Institute of Pathophysiology, Ljubodrag Buba Mihailović, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Bojan Jorgačević
Affiliation:
Institute of Pathophysiology, Ljubodrag Buba Mihailović, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Danijela Vučević
Affiliation:
Institute of Pathophysiology, Ljubodrag Buba Mihailović, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
Tatjana Radosavljević*
Affiliation:
Institute of Pathophysiology, Ljubodrag Buba Mihailović, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia
*
*Author for correspondence: Tatjana Radosavljević, E-mail: [email protected]
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Abstract

The effects of betaine on hepatocytes chromatin architecture changes were examined by using fractal and gray-level co-occurrence matrix (GLCM) analysis in methionine/choline-deficient (MCD) diet-induced, nonalcoholic fatty liver disease (NAFLD). Male C57BL/6 mice were divided into groups: (1) Control: standard diet; (2) BET: standard diet and betaine supplementation through drinking water (solution 1.5%); (3) MCD group: MCD diet for 6 weeks; (4) MCD+BET: fed with MCD diet + betaine for 6 weeks. Liver tissue was collected for histopathology, immunohistochemistry, and determination of fractal dimension and GLCM parameters. MCD diet induced diffuse micro- and macrovesicular steatosis accompanied with increased Ki67-positive hepatocyte nuclei. Steatosis and Ki67 immunopositivity were less prominent in the MCD+BET group compared with the MCD group. Angular second moment (ASM) and inverse difference moment (IDM) (textural homogeneity markers) were significantly increased in the MCD+BET group versus the MCD group (p<0.001), even though no difference between the MCD and the control group was evident. Heterogeneity parameters, contrast, and correlation were significantly increased in the MCD group versus the control (p<0.001). On the other hand, betaine treatment significantly reduced correlation, contrast, and entropy compared with the MCD group (p<0.001). Betaine attenuated MCD diet-induced NAFLD by reducing fat accumulation and inhibiting hepatocyte proliferation. Betaine supplementation increased nuclear homogeneity and chromatin complexity with reduction of entropy, contrast, and correlation.

Type
Biological Science Applications
Copyright
© Microscopy Society of America 2018 

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Footnotes

Cite this article: Vesković M, et al. (2018). The Effects of Betaine on the Nuclear Fractal Dimension, Chromatin Texture, and Proliferative Activity in Hepatocytes in Mouse Model of Nonalcoholic Fatty Liver Disease. Microscopy and Microanalysis. doi: 10.1017/S1431927617012806

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