Hepatic transcriptome and proteome analyses provide new insights into the regulator mechanism of dietary avicularin in diabetic mice.

Many dietary flavonoids existing as glycosides in fruits and vegetables are considered bioactive food components with various potential health benefits. Type 2 diabetes mellitus (T2DM) is a complex and polygenic disease with increasing global prevalence and economic burden. In this study, the hypoglycemic effect of avicularin (quercetin-3-O-α-arabinofuranoside), a flavonoid glycoside commonly found in natural plants and fruits, was determined in a high fat diet/streptozotocin induced type 2 diabetes mouse model. Our results demonstrated that dietary avicularin treatment reduced levels of fasting blood glucose, serum TG and LDL-C, liver AST and ALT, and increased hepatic glycogen in T2DM mice. Furthermore, we used RNA-Seq and iTRAQ to compare the gene and protein expression in the livers of the normal control mice (NC), diabetic control mice (DC) and avicularin treated mice (DA100). The differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were analyzed based on gene annotations and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Integrated analysis of the RNA-Seq and iTRAQ data indicated that the fifteen DEGs/DEPs showed the same trend in mRNA and protein expression levels in comparisons of both NC vs DC and DC vs DA100. KEGG analysis revealed that four DEGs/DEPs (PKM, PEPCK, PYG, and PLA2) in the glycolysis, gluconeogenesis, and arachidonic acid pathway, and six DEPs (Ndufb4, Ndufa6, Cox5a, Cox5b, Cox6c, and ATPSß) in the oxidative phosphorylation signaling pathway, play important roles in avicularin's hypoglycemic effect. We also found six other DEGs/DEPs related to T2DM (CA1, Serpinb6a, AK, Pcolce, Cand2, and Atp2a3), and five related to cancer (Phgdh, Tes, Papss1, Psat1, and Fam49b). We did further verify by qRT-PCR and explored the possible binding modes of avicularin with targeted proteins with molecular docking simulations. Taken together, our results demonstrated the protective effects of avicularin against diabetes and provided a global view about the system-level hypoglycemic mechanisms of avicularin by the comprehensive analysis of transcriptomic and proteomic data in T2DM mice.

Effects of casein phosphopeptides on calcium absorption and metabolism bioactivity in vitro and in vivo.

Casein phosphopeptides (CPPs) are a series of peptides containing serine phosphate, which prevents calcium precipitation in the small intestine, so that it can be absorbed. Calcium metabolism studies have been typically carried out using traditional metabolic balance tests. Non-radioactive stable isotopes have rarely been used to examine the in vivo absorption and metabolism of calcium in animal models due to their high cost and the difficulty in their accurate detection. However, they have recently attracted intense research attention. In this study, we developed InertSep ME-1 using a chelating resin to effectively separate 42Ca from spectral-interference in the process of ICP-MS detection. The method effectively removes interfering ions like potassium and sodium and greatly improves the accurate detection of the calcium ion isotope. We also investigated the absorption, distribution, and metabolism of different CPPs through both in vitro cell experiments and in vivo animal experiments. The results indicate that CPPs strongly promote calcium absorption, especially the P5 active monomer component. The results of our in vivo studies show that the calcium isotope can be absorbed from the small intestine into the blood. Then, one part is transported to various organs through tissue fluids while another part is excreted into the urine through the kidneys. In general, our results reveal that CPPs promote the absorption of calcium significantly and positively affect calcium metabolism.