Crocetin ameliorates non-alcoholic fatty liver disease by modulating mitochondrial dysfunction in L02 cells and zebrafish model.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese medicine considers that the etiology and pathogenesis of non-alcoholic fatty liver disease (NAFLD) are related to liver depression and qi stagnation. Saffron and its active ingredient, crocetin (CCT), are used for the treatment of metabolic diseases owing to their "Liver deobstruent" and "Liver tonic" effects. However, the effect of CCT on NAFLD has not been fully elucidated. In the present study, the effect and potential molecular mechanism of CCT were explored in both in vivo and in vitro models of NAFLD. MATERIALS AND METHODS: CCT was isolated from saffron and purity and structure characterization were performed using HPLC, MS, 1H-NMR, and 13C-NMR. The effect of CCT on the viability of L02 cells and its maximum tolerable concentration (MTC) in zebrafish were investigated. Free fatty acids (FFA) and thioacetamide (TAA) were used to induce lipid accumulation in L02 cells and steatosis in zebrafish, respectively. The effects of CCT on indexes related to lipid metabolism, oxidative stress, and mitochondrial function in NAFLD models were explored using biochemical assay kits, Western blot analysis, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), histopathology analysis, and determination of mitochondrial membrane potential (ΔΨm). Morphological analysis of mitochondria was performed using transmission electron microscopy (TEM). RESULTS: The levels of triglyceride (TG), total cholesterol (TC), malondialdehyde (MDA), and alanine/aspartate aminotransferases (ALT/AST) activities in FFA treated L02 cells were significantly reduced after CCT treatment. CCT treatment significantly increased ATP concentration, ΔΨm, and activities of superoxide dismutase (SOD), catalase (CAT), and cytochrome c oxidase (COX IV) in FFA treated L02 cells. TEM images showed restoration of mitochondrial morphology. CCT decreased ATP concentration and upregulated expression of B-cell lymphoma-2 (Bcl-2) and COX IV, whereas, CCT downregulated expression of BCL2-Associated X (Bax) and cleaved caspase-3 in TAA treated zebrafish. These findings indicated that mitochondrial dysfunction was alleviated after CCT treatment. Oil Red O staining of L02 cells and zebrafish showed that CCT treatment reversed the accumulation of lipid droplets. CONCLUSION: In summary, CCT treatment effectively alleviated the symptoms of NAFLD and restored mitochondrial function in L02 cells and zebrafish NAFLD model.

Effect of crocin on the insulin resistance and lipid profile of streptozotocin-induced diabetic rats.

Crocin is the only water soluble carotenoid in nature, and it has a known powerful antioxidant activity. The aim of this work was to investigate the hypoglycemic and hypolipidemic effects of crocin in streptozotocin (STZ)-induced type 2 diabetic rats. Neonatal male Wistar rats (2-5 days old) were randomly divided into five groups. Three groups were intraperitoneally injected with STZ (90 mg/kg body weight). Among them, two groups were treated with intraperitoneal injection of crocin (50 or 100 mg/kg), and the third group was treated with vehicle only. Two control groups were also considered, and one of them was treated with crocin. After 5 months, their blood and urine samples were collected, and the animals were sacrified. The results indicate a significant lower body weight (P < 0.001) and abnormal parameters in the diabetic rats compared with the normal group. An administration of both doses of crocin significantly decreased the levels of serum glucose, advanced glycation end products, triglyceride, total cholesterol, and low-density lipoprotein and increased the high-density lipoprotein in the diabetic rats. The treatments were also effective in decreasing HbA1c and microalbuminuria, as well as homeostatic model assessment for insulin resistance as a measure of insulin resistance in the diabetic rats.