The oligosaccharides extracted from the seeds of peas, specifically consisting of raffinose, stachyose, and verbascose, fall under the category of raffinose family oligosaccharides (RFOs). The effect of RFOs on intestinal microflora and the anti-inflammatory mechanism were investigated by in vitro fermentation and cell experiments. Firstly, mouse feces were fermented in vitro and different doses of RFOs (0~2%) were added to determine the changes in the representative bacterial community, PH, and short-chain fatty acids in the fermentation solution during the fermentation period. The probiotic index was used to evaluate the probiotic proliferation effect of RFOs and the optimal group was selected for 16S rRNA assay with blank group. Then, the effects of RFOs on the inflammatory response of macrophage RAW264.7 induced by LPS were studied. The activity of cells, the levels of NO, ROS, inflammatory factors, and the expression of NF-κB, p65, and iNOS proteins in related pathways were measured. The results demonstrated that RFOs exerted a stimulatory effect on the proliferation of beneficial bacteria while concurrently inhibiting the growth of harmful bacteria. Moreover, RFOs significantly enhanced the diversity of intestinal flora and reduced the ratio of Firmicutes-to-Bacteroides (F/B). Importantly, it was observed that RFOs effectively suppressed NO and ROS levels, as well as inflammatory cytokine release and expression of NF-κB, p65, and iNOS proteins. These findings highlight the potential of RFOs in promoting intestinal health and ameliorating intestinal inflammation.

2’-Fucosyllactose (2’-FL), the functional oligosaccharide naturally present in milk, has been shown to exert health benefits. This study was aimed to investigate the effect of 2’-fucosyllactose (2’-FL) on the browning of white adipose tissue in 3T3-L1 adipocytes and C3H10T1/2 cells. The results revealed that 2’-FL decreased lipid accumulations with reduced intracellular triglyceride contents in vitro. 2’-FL intervention increased the mitochondria density and the proportion of UCP1-positive cells. The mRNA expressions of the mitochondrial biogenesis-related and browning markers (Cox7a, Cyto C, Tfam, Ucp1, Pgc1α, Prdm16, Cidea, Elovl3, Pparα, CD137, and Tmem26) were increased after 2’-FL intervention to some extent. Similarly, the protein expression of the browning markers, including UCP1, PGC1α, and PRDM16, was up-regulated in the 2’-FL group. Additionally, an adenosine monophosphate-activated protein kinase (AMPK) inhibitor, compound C (1 μM), significantly decreased the induction of thermogenic proteins expressions mediated by 2’-FL, indicating that the 2’-FL-enhanced beige cell formation was partially dependent on the AMPK pathway. In conclusion, 2’-FL effectively promoted the browning of white adipose in vitro.

Huai chrysanthemum (Chrysanthemum morifolium Ramat. cv. Huaiju), collected from Jiaozuo, Henan Province, China, is a medicinal and edible plant rich in phenolic compounds. Polyphenols are natural bioactive compounds with potential antibacterial activity. In this study, HPLC-MS/MS was used to identify and quantify the phenolic compounds in huai chrysanthemum polyphenol extract, and its antibacterial activity against representative Gram-negative and Gram-positive foodborne bacteria was evaluated. A total of 22 polyphenolic compounds were identified, including 10 phenolic acids, 10 flavonoids, and two other phenolic compounds. Caffeic acid was the predominant phenolic acid, while luteoloside was the predominant flavonoid. Flavonoids accounted for 94.03% of the total identified polyphenols. The extract inhibited the growth of both Escherichia coli and Staphylococcus aureus, with an MIC₈₀ of 12.5 mg/mL for both strains. Further analysis showed that the extract reduced intracellular protein content and ATP levels, increased extracellular alkaline phosphatase activity, altered cell surface charge, affected superoxide dismutase activity, and caused marked morphological changes in bacterial cells. These results suggested that the antibacterial effect of huai chrysanthemum polyphenol extract was related to cell envelope damage, energy metabolism disturbance, and redox imbalance. This study provided a basis for further application of huai chrysanthemum polyphenol extract as a natural antibacterial ingredient in food preservation.