Background: Fatty liver is the most prevalent chronic liver disease. The progressive form of fatty liver disease is non-alcoholic steatohepatitis (NASH), which can lead to liver fibrosis and cirrhosis. Silymarin is a traditional liver treatment while Mesenchymal stem cells conditioned medium (MSCs-CM) is a new hope for treatment of chronic liver diseases. Our aim is to evaluate the effect of MSCs-CM versus Silymarin on experimentally induced fatty liver in adult male albino rats. Methods: Fifty-four adult male albino rats were divided into four groups: I (control group 27 rats) divided into 3 subgroups: (Subgroup Ia, Ib and Ic), II (fatty liver group), III (fatty liver -Silymarin) and IV (fatty liver -MSC-CM). Blood samples were collected for lipid profile. Liver tissue was prepared for H&E staining, Mallory's trichrome staining, osmic acid staining, immunohistochemically staining for alpha smooth muscle actin (α-SMA) and caspase 3 and electron microscope examination. Statistical and morphometrical studies were performed. Results: Regarding lipid profile, α-SMA and caspase-3 the highest mean values were observed in group II while the least in group IV. H&E-stained sections of group II showed degenerated hepatocytes with vacuolated cytoplasm and darkly stained nuclei. Ultra-structurally, group II revealed, rarified cytoplasm with swollen mitochondria and bundles of collagen while in group III showed some improvement, group IV most of hepatocytes appeared normal. Conclusions: MSCs-CM has a more ameliorative effect than Silymarin on induced non-alcoholic fatty liver disease through its anti-apoptotic and anti-inflammatory effects.