Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder characterized by the accumulation of fat in the liver without alcohol consumption. NAFLD is a significant public health concern worldwide due to its increasing prevalence and potential progression to non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma (HCC). Various dietary and lifestyle factors, such as high-fat diets, sedentary lifestyle, and obesity, contribute to the development and progression of NAFLD. Currently, there is no effective pharmacological therapy for NAFLD, and lifestyle modifications such as weight loss and increased physical activity remain the mainstay of treatment. Natural products have been explored as alternative approaches for the modulating of NAFLD. Raspberry ketone (RK) and black seed oil (NS) are natural compounds that have shown promise in reducing the risk of metabolic disorders such as obesity, insulin resistance, and dyslipidemia. However, their potential therapeutic effects on NAFLD are yet to be fully elucidated. Aims of the study 1- To elucidate the relation between TLR4, SphK1 in NAFLD and its metabolic / inflammatory changes. 2- To evaluate the effect of diet, RK and/or NS on sphingosine kinase/sphingosine-1-phosphate and TLR4 pathways in NAFLD. The rats were divided into seven groups: normal control (NC), NAFLD (induced by HFFD), NAFLD rats shifted to normal diet (ND), calorie restricted (CR) group, RK group, NS group, and RK+NS combination group. The rats were fed with different diets and treated with RK, NS, or both for eight weeks. Body weight was reported weekly. After 14 weeks, blood and liver tissues samples were collected for biochemical and histological analyses. Serum levels of ALT, AST, TC, TAG, and HDL-C, were measured using standard diagnostic kits and non HDL-C was calculated according to the following formula: Non HDL-C (mg/dl) = TC – HDL-C Hepatic oxidative stress, inflammatory markers, and sphingosine pathway parameters were measured using relevant ELISA kits. Histopathological examination of liver tissues was performed by hematoxylin and eosin (HE) staining and oil red O (ORO) staining. Results this study revealed that NAFLD rats had significantly higher body and liver weights, as well as elevated serum levels of total cholesterol (TC), triglycerides (TAGs), and non high-density lipoprotein cholesterol (non HDL-C) and decreased high-density lipoprotein cholesterol (HDL-C) compared to normal control rats. NAFLD rats also had increased hepatic expression of sphingosine kinase 1 (SphK1), sphingosine-1-phosphate (S1P), and sphingosine-1-phosphate receptor 1 (S1PR1), as well as tumor necrosis factor alpha (TNF-α), toll-like receptor 4 (TLR4), and nuclear factor kappa B (NF-κB). They also had decreased antioxidant activities, as indicated by lower hepatic content of superoxide dismutase (SOD) and reduced glutathione (GSH). Histopathological examination of liver tissue showed hepatocyte damage and lipid accumulation in NAFLD rats. However, rats switched to a calorie-restricted diet (CR) or treated with raspberry ketone (RK) along with normal calorie diet showed improvements in body and liver weights, serum lipid levels, hepatic expression of SphK1, S1P, and S1PR1, TNF-α, TLR4, and NF-κB, as well as antioxidant activities. Additionally, the RK and NS groups showed a significant increase in tissue content of TLR4, S1P, and S1PR1 compared to the combination group (NS+RK). The NS+RK combination group showed a significant reduction in tissue content of SphK1 compared to the CRD group. Histopathological examination of liver tissue also showed reduced lipid accumulation and improved liver architecture in CR and RK-treated rats. The results suggest that RK and CR may have potential therapeutic effects on NAFLD by improving various tissue parameters. The study also found that RK treatment was more effective than CR alone in reducing lipid accumulation in the liver. Conclusion The present study highlights the potential of dietary interventions, such as raspberry ketone (RK), black seed oil (NS), and their combination NS+RK, as well as a calorie-restricted diet (CRD), in reducing weight gain, dyslipidemia, inflammation, and oxidative stress in a rat model of non-alcoholic fatty liver disease (NAFLD). The findings suggest that the dysregulation of the SphK1/S1P signaling pathway may contribute to the pathogenesis of NAFLD, and dietary interventions may play a role to attenuate this dysregulation. The results also suggest that the combination of NS+RK may have a synergistic effect in regulating S1P-related signaling pathways, and TLR-4 pathway along with antioxidant defense mechanisms. Recommendation These findings provide important insights into the potential of natural compounds (e.g raspberry ketone and black seed oil) as complementary agents for attenuating NAFLD complications. This may need further experimental and clinical trials on a large scale to make use of these results.