The aim of this study was to investigate the antihypercholesterolemic activity and potential molecular mechanism of columbamine (COL) that was prepared by extraction from Rhizoma Coptidis in hamsters and HepG2 cells. The results displayed that the COL from Rhizoma Coptidis was a safe natural compound with a LD50 0f 1524.6mg/kg and no detectable toxic symptoms during the observation of chronic toxicity. COL dose-dependently reversed the abnormal lipid levels induced by HFHC diet. Specifically, COL(M) and COL(H) significantly reduced the blood lipid levels(TC, TG and LDL-c) and enhanced the fecal contents of TBA by 21.8% and 25.1% respectively in hamsters. COL up-regulated the genes of CYP8B1, CYP7A1 and LDLR in mRNA and protein level, and down-regulated those of HMGCR to a different degree. Especially, CYP7A1 were significantly up-regulated by COL in hamsters (p<0.01). Further analysis indicated that COL obviously activated the mRNA and protein expression of the transcription factors FTF, HNF-4α, and inhibited those of SHP. Promoter luciferase assay showed that COL induced the expression of FTF and HNF-4α, further transactivating CYP7A1, which accelerated the conversion of liver cholesterol to bile acids. It concluded that the COL showed high lipid-lowering activities through indirectly transactivating CYP7A1 by upregulating FTF and HNF-4α, and directly activating CYP7A1 catalytic activity by strongly interacting with receptor and ligand, therefore promoting cholesterol catabolism and accelerating the excretion of bile acids.
Keywords: CYP7A1; Cholesterol (PubChem CID:5997); Columbamine; Columbamine (PubChem CID:72310); Creatinine (PubChem CID:588); Dimethyl sulfoxide (PubChem CID:679); FTF; HNF-4α; Hypercholesterolemia; Methylthio-tetrazole (PubChemCID:97664); Paraformaldehyde (PubChem CID:712); Penicillin (PubChem CID:5904); Simvastatin (PubChem CID:54454); Streptomycin (PubChem CID:19649); Triglyceride (PubChem CID:5322095).
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