Investigation of the Therapeutic Effect of Silymarin, the Silybum marianum Medicinal Plant Extract, in an Empirically Induced in Vitro Model of Non-Alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD) is a common digestive disorder that can cause some additional liver problems. Nowadays developing efficient therapies for the prevention and control of hepatic diseases is crucial. Recently, replacing synthetic drugs with natural herbal remedies has been more favored due to their ever-growing demand and public acceptance. Our goals in this study were the experimental induction of non-alcoholic fatty liver disease in the hepatocyte cell line and the evaluation of the therapeutic efficiency of silymarin, the Silybum marianum medicinal plant extract. The HepG2 cells were cultured in DMEM/F12 medium supplemented with 10% fetal bovine serum and 2 mM GlutaMAX. The effect of various concentrations of fructose in the induction of the fatty liver condition was assessed. Subsequently, the diseased hepatocytes were treated with different concentrations of silymarin. The therapeutic effect of silymarin was evaluated by fluorescent microscopy of the cells stained with a lipid-specific fluorescent dye and by quantifying the hepatic enzymes. Cultivation of HepG2 cells for seven days in the medium containing 30 mM fructose was found appropriate in the induction of the NAFLD condition. The lipid-specific staining, as well as the measurement of the hepatic enzymes, revealed the therapeutic efficiency of the 200 and 250 μM concentrations of silymarin. The experiments showed that Nile Red stain, silymarin, and DMSO were not notably cytotoxic. A high-energy diet provided by the added fructose in the culture medium was effective in resembling the NAFLD condition in HepG2 cells in Vitro. Furthermore, silymarin was found to be effective in curing the NAFLD condition in a time and dose-dependent manner.