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HCC (Hepatocellular Carcinoma) is a critical health issue worldwide. Our previous animal experiment has confirmed that blueberry malvidin-3-galactoside (M3G) can regulate the progression of HCC. In this study, feces samples from the same batch of mice were collected to explore the regulatory mechanism of M3G on intestinal microbiota and microbial TCA cycle metabolism KEGG pathway in HCC mice based on 16S rRNA sequencing and metagenomics. Our results showed that blueberry M3G increased the microbial diversity and regulated the structure of intestinal microbiota in mice, such as increasing the abundance of Clostridia (butyric acid-producing bacteria), Oscillospira and Ruminococcus, and reducing the abundance of pathogenic Erysipelotrichi. Compared with the group of liver cancer (LC) and 5-fluorouracil (5 F), blueberry M3G significantly regulated microbial TCA cycle KEGG pathway via improving the expression of key proteins (porA, DLAT, aceE, PC and OGDH). Additionally, we found which the abundance of Muribaculum intestinale (M.intestinale) increased by blueberry M3G may be an important factor affecting the microbial TCA cycle KEGG pathway via the pearson correlation (R) analysis of protein and microbiota. Taken together, these results demonstrate that the blueberry M3G has the potential to be an intestinal microbiota regulator and an adjuvant to HCC therapy.

Malvidin-3-galactoside (Mv-3-gal) is the major anthocyanin monomer in blueberry anthocyanins. The compound is well-characterized by its anti-oncogenesis function in multiple organs including liver. In the current study, the mechanism driving the anti-hepatocellular carcinoma (HCC) function of Mv-3-gal was further explored by focusing on apoptosis and metastases pathways. HCC cell line Huh-7 was firstly administrated with Mv-3-gal of different concentrations and the effect of Mv-3-gal on cell proliferation, apoptosis, colony formation ability, metastasis potential as well as the activity of Akt/PTEN and MAPK pathways was assessed. The results showed that Mv-3-gal inhibited the cell proliferation and colony formation ability, induced cell cycle arrest and apoptosis in a dose-dependent manner. Regarding the metastasis potential, Mv-3-gal suppressed the migration and invasion potential of Huh-7 cells by regulating MMPs expression. Taken together, the findings highlighted the anti-HCC potent of Mv-3-gal, which was associated with the inhibition of Akt/PTEN, MAPK and MMP pathways.

Proteins and anthocyanins coexist in complex food systems. This research mainly studied the steady-state protective design and mechanism of the preheated protein against anthocyanins. Multispectral and molecular dynamics are utilized to illustrate the interaction mechanism between preheated whey protein isolate (pre-WPI) and anthocyanins. The pre-WPI could effectively protect the stability of anthocyanins, and the effect was better than that of the natural whey protein isolate (NW). Among them, NW after preheating treatment at 55 °C showed better protection against anthocyanin stability. Fluorescence studies indicated that pre-WPI there existed a solid binding affinity and static quenching for malvidin-3-galactoside (M3G). Multispectral data showed a significant variation in the secondary structure of pre-WPI. Furthermore, molecular dynamics simulation selects AMBER18 as the protein force field, and the results showed that hydrogen bonding participated as an applied force. Compared with NW, pre-WPI could better wrap anthocyanins and avoid damage to the external environment due to tightening of the pockets. Protein protects anthocyanins from degradation, and this protective effect is influenced by the preheating temperature of protein and the structure of protein. On the basis of the above results, it is possible to pinpoint the interaction mechanism between preheated proteins and anthocyanins. [ABSTRACT FROM AUTHOR]

Blueberry fruits have a wide range of health benefits because of their abundant anthocyanins, which are natural antioxidants. The purpose of this study was to investigate the inhibitory effect of blueberry’s two main anthocyanins (malvidin-3-glucoside and malvidin-3-galactoside) on inflammatory response in endothelial cells. These two malvidin glycosides could inhibit tumor necrosis factor-alpha (TNF-α) induced increases of monocyte chemotactic protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) production both in the protein and mRNA levels in a concentration-dependent manner. Mv-3-glc at the concentration of 1 μM could inhibit 35.9% increased MCP-1, 54.4% ICAM-1, and 44.7% VCAM-1 protein in supernatant, as well as 9.88% MCP-1 and 48.6% ICAM-1 mRNA expression (p < 0.05). In addition, they could decrease IκBα degradation (Mv-3-glc, Mv-3-gal, and their mixture at the concentration of 50 μM had the inhibition rate of 84.8%, 75.3%, and 43.2%, respectively, p < 0.01) and block the nuclear translocation of p65, which suggested their anti-inflammation mechanism was mediated by the nuclear factor-kappa B (NF-κB) pathway. In general malvidin-3-glucoside had better anti-inflammatory effect than malvidin-3-galactoside. These results indicated that blueberry is good resource of anti-inflammatory anthocyanins, which can be promising molecules for the development of nutraceuticals to prevent chronic inflammation in many diseases.