Mechanism of crocin I on ANIT-induced intrahepatic cholestasis by combined metabolomics and transcriptomics.

Background: Intrahepatic cholestasis (IC) is a disorder of bile production, secretion, and excretion with various causes. Crocin I (CR) is effective in the treatment of IC, but its underlying mechanisms need to be further explored. We aimed to reveal the therapeutic mechanism of crocin I for IC by combining an integrated strategy of metabolomics and transcriptomics. Methods: The hepatoprotective effect of CR against cholestasis liver injury induced by α-naphthylisothiocyanate (ANIT) was evaluated in rats. The serum biochemical indices, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acid (TBA), total bilirubin (TBIL), direct bilirubin (DBIL), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), and interleukin 1ß (IL-1ß), as well as the liver oxidative stress indexes and the pathological characteristics of the liver were analyzed. In addition, we also performed a serum metabolomics study using UPLC-Q Exactive HF-X technology to investigate the effect of CR on the serum of rats with ANIT-induced IC and screened potential biomarkers. The enrichment analysis of differential expressed genes (DEGs) was performed by transcriptomics. Finally, the regulatory targets of CR on potential biomarkers were obtained by combined analysis, and the relevant key targets were verified by western blotting. Results: CR improved serum and liver homogenate indexes and alleviated liver histological injury. Compared with ANIT group, the CR group had 76 differential metabolites, and 10 metabolic pathways were enriched. There were 473 DEGs significantly changed after CR treatment, most of which were enriched in the retinol metabolism, calcium signaling pathway, PPAR signaling pathway, circadian rhythm, chemokine signaling pathway, arachidonic acid metabolism, bile secretion, primary bile acid biosynthesis, and other pathways. By constructing the "compound-reaction-enzyme-gene" interaction network, three potential key-target regulation biomarkers were obtained, including 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), ATP-binding cassette transporter G5 (ABCG5), and sulfotransferase2A1(SULT2A1), which were further verified by western blotting. Compared with the ANIT group, the CR group significantly increased the expression of ABCG5 and SULT2A1, and the expression of HMGCR significantly decreased. Conclusion: Combined metabolomic and transcriptomic analyses show that CR has a therapeutic effect on IC through regulation of the biosynthesis of bile acids and bilirubin in the bile secretion pathway and regulation of the expression of HMGCR, ABCG5, and SULT2A1.

[Comparative Study on Protection Effects of CCl4-Induced Acute Liver Injury of Different Prepared Products from Gardenia jasminoides Fruits in Rats].

Objective: To provide the experimental evidence for the appropriate selection of the different prepared products from Gardenia jasminoides fruits by comparing their protection effects on carbon tetrachloride（ CCl4）-induced acute liver injury. Methods: The activities of ALT,AST,ADA,LDH,ALP and contents of PA,TP,TBIL,DBIL,TBA in serum,the activities of SOD and the content of MDA in liver tissue were measured in acute liver injury rats by carbon tetrachloride. Also the pathological changes of liver tissues were examined under microscope. Results: The biochemical indexes of AST,ALT,TBA,ADA,LDH and MDA were significantly improved in all groups of prepared products from Gardenia jasminoides fruits,but not SOD and ALP. The lesions of liver tissue had different degrees of reduction. Conclusion: The different prepared products from Gardenia jasminoides fruits had the effects of liver protection. The nut of Gardeniae Fructus was superior to the peel in enzyme decreasing and liver protection. The crude was superior to the stir-cooked in enzyme decreasing and liver protection.

[Study on chemical constituents of Gardenia jasminoides(III)].

OBJECTIVE: To investigate the chemical constituents of Gardenia jasminoides fruits. METHODS: Various column chromatography were used in the isolation and purification, and physiochemical constant determination and spectral analysis were adopted to determine the chemical structures. RESULTS: Twelve compounds were isolated from Gardenia jasminoides including jasminoside I (1), gardenoside (2), gardaloside (3), 3-hydroxy-urs-12-ene-11-ketone(4), 5, 4'-dihydroxyl-7, 3', 5'-trimethoxyflavone (5), 5, 7, 3', 4', 5'-pentamethoxyflavone(6), 3, 5, 6, 4'-tetrahydroxy-3', 5'-dimethoxyflavone (7), shikimic acid (8), 1, 2, 4-benzenetriol (9), 3, 4-dimethoxy-benzoic acid (10), dibutyl phthalate (11) and diisobutyl phthalate (12). CONCLUSION: Compounds 4 - 7 and 9 -10 were isolated from this plant for the first time.

[Study on the chemical components of Gardenia jasminoides (II)].

OBJECTIVE: To investigate the chemical components of Gardenia jasminoides. METHODS: Various column chromatography were used in the isolation and purification, and physiochemical constant determination and spectral analysis were adopted to idenitify the chemical structures. RESULTS: Ten compounds were isolated and identified as jasminoside A(1), epijasminoside A(2), 6-O-methylscandoside methyl ester (3), 6-O-methyldeacetylasperulosidic acid methyl ester (4), gardenoside (5), phenylmethol (6), 4-hydroxy-phenylmethol-O-beta-D-glucopyranosyl- (1-->6) -beta-D-glucopyranoside (7), 3,4-dihydroxy-phenylmethol-O-beta-D-glucopyranosyl-(1-6)-beta-D-glucopyranoside (8), 3-hydroxy4-methoxy-phenylmethol-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside (9), 3-hydroxy-4-methoxyphenylmethol-O-beta-D-glucopyranoside (10). CONCLUSION: Compounds 6 -10 are isolated from this plant for the first time.