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Evaluating the effect of a mixture of two main conjugated linoleic acid isomers on hepatic steatosis in HepG2 cellular model

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Abstract

Hepatic steatosis is an early form of non-alcoholic fatty liver disease (NAFLD), caused by abnormal fat deposition in the hepatocytes. Conjugated linoleic acid (CLA) is a group of positional and geometric dienoic isomers of linoleic acid that attract significant attention because of its beneficial effects on chronic diseases such as cancer, obesity, and metabolic syndrome. This study examined the influence of a mixture of two main CLA isomers (CLA-mix) on lipid accumulation and lipid metabolism-related genes using HepG2 cells treated with palmitic acid (PA) as an in vitro model for hepatic steatosis. Methods and Results: HepG2 cells were treated for 24 h: control (BSA), model (BSA + PA), and treated groups (BSA-PA + non-toxic concentrations of CLA-mix). Intracellular lipid deposition, triglyceride (TG), total cholesterol (TC) and gene expression were measured by Oil-Red O staining, colorimetric assay kits and real-time PCR, respectively. CLA-mix at high concentrations had significantly decreased intracellular total lipid and TG deposition compared to the model group. However, none of the CLA-mix concentrations had a significant effect on the intracellular TC level. CLA-mix significantly increased the expression of some genes mainly regulated by PPARα but did not alter the expression of lipogenesis-related genes. Conclusions: These results demonstrate that high concentrations of CLA-mix protect against hepatic steatosis and play a role in regulating fatty acid oxidation and bile excretion through the PPARα pathway. It is suggested that the effect of different ratios of two main CLA isomers on the amount and ratio of bile compounds be investigated in future studies.

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Data availability

Data of this study are included in the article and the primary data can be provided from the corresponding author.

Abbreviations

ACC:

Acetyl-CoA carboxylase

AA:

Arachidonic acid

ABCG8:

ATP binding cassette subfamily G member 8

BA:

Bile acid

BSEP:

Bile salt export pump

BSA:

Bovine serum albumin

CPT-1:

Carnitine palmitoyltransferase-I:

Chol:

Cholesterol

CAA:

Conjugated arachidonic acid

CLA:

Conjugated linoleic acid

FXR:

Farnesoid X receptor

FABP:

Fatty acid-binding protein

FAS:

Fatty acid synthase

FFA:

Free fatty acid

Glc:

Glucose

Gly:

Glycerol

HMGCR:

3-Hydroxy-3methyl-glutaryl-coenzyme A reductase

HMGCS2:

3-Hydroxy-3-methylglutaryl-coa synthase 2

LA:

Linoleic acid

LPL:

Lipoprotein lipase

LXRα:

Liver X receptor alpha

MDR3:

Multidrug resistance protein 3

NAFLD:

Non-alcoholic fatty liver disease

NASH:

Non-alcoholic steatohepatitis

OA:

Oleic acid

PA:

Palmitic acid

ACOX1:

Peroxisomal acyl-coenzyme A oxidase1

α:

β, γ1, γ2, Peroxisome proliferator-activated receptors

PL:

Phospholipid

SA:

Stearic acid

SREBP-1c:

Sterol-regulatory element binding protein-1c

TG:

Triglyceride

TC:

Total cholesterol

UCP:

Uncoupling protein

VLDL:

Very low density lipoprotein

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Acknowledgements

This experimental study was part of a Ph.D. program and supported by a grant from the Tehran University of Medical Sciences (TUMS Grant Number: 96-02-30-35441).

Funding

This project was financially supported by grant (TUMS Grant Number: 96-02-30-35441) from the Deputy of Research, Tehran University of Medical Sciences.

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Ali Jalilian, Taghi Golmohammadi, Reza Meshkani, Neda Eivazi, Reyhaneh Babaei Khorzoughi, Seyed Reza Hosseini fard & Maliheh Paknejad

  2. Department of Clinical Biochemistry, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Mehdi Koushki

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  1. Ali Jalilian
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Contributions

AJ, MP and TG designed the project. AJ carried out all experiments and wrote the manuscript. MP provided all the required materials and advice about this study. TG provided valuable instructions and suggestions for this study. RM Helpful and effective tips during project implementation. NE helped for cell culture and optimizing of MTT assay. RB helped for optimizing of Oil Red assay and TG & TC measurements. MK helped for optimizing real-time PCR and analysis of data. SRH helped in editing the manuscript and designing the schematic image. All authors read and approved the final manuscript.

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Correspondence to Taghi Golmohammadi or Maliheh Paknejad.

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Jalilian, A., Golmohammadi, T., Meshkani, R. et al. Evaluating the effect of a mixture of two main conjugated linoleic acid isomers on hepatic steatosis in HepG2 cellular model. Mol Biol Rep 48, 1359–1370 (2021). https://doi.org/10.1007/s11033-021-06203-5

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