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.

[A new antioxidant flavonoid glycoside from flowers of Hosta plantaginea].

Phytochemical investigation of the flowers of Hosta plantaginea led to isolate of one new flavonoid glycoside,plantanone C( 1) by silica gel,Sephadex LH-20,and RP-HPLC column chromatographies. Its structure was extensively determined on basis of HR-ESI-MS and NMR spectroscopic data. Compound 1 exhibited moderate antioxidant activity against DPPH radical scavenging activity,with an IC50 value of 240. 2 µmol·L~(-1).

Anti-inflammatory and antioxidant activities of flavonoids from the flowers of Hosta plantaginea.

Hosta plantaginea was a traditional Chinese medicinal plant used to treat inflammation-related diseases with little scientific validation. Twelve flavonoids, including two new compounds namely plantanones A (1) and B (2), were isolated from the flowers of Hosta plantaginea. Their structures were elucidated by NMR and HRMS as well as comparison with literature data. All of the isolated compounds showed significant inhibitory activities against ovine COX-1 and COX-2 at a concentration of 50 µM, with inhibition ratios from 53.00% to 80.55% for COX-1 and from 52.19% to 66.29% for COX-2. Further detailed testing showed that compounds 1, 2, 4 and 12 inhibited the COX-1 and COX-2 enzymes with IC50 values 12.90-33.37 µM and 38.32-46.16 µM, respectively. Moreover, the antioxidant effects of these isolates against DPPH free radical-scavenging were also evaluated in vitro, and a tight structure-activity relationship was discussed. Our results suggested that the anti-inflammatory and antioxidant activities of H. plantaginea flowers are partly attributed to these flavonoids.