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The objective of the current study was to compare the abilities of undigested and enzymatically digested bifidobacteria toinduce nitric oxide and cytokine release in RAW 264.7 macrophage cells. Nine different Bifidobacterium strains derivedfrom herbivorous animals were digested with pepsin and then pancreatin, and the precipitates and supernatants wereacquired via centrifugation. The RAW 264.7 cells were incubated with whole cells, the precipitate, or the supernatant, andthe macrophage culture supernatants were analyzed with respect to the induction of nitric oxide and cytokines. Pronouncedincreases in the production of nitric oxide, interleukin (IL)-1β, IL-6, IL-12, and tumor necrosis factor-α (TNF-α) wereobserved when cultured with whole cells and the precipitates. It is noteworthy that the precipitates in most of the Bifidobac-terium strains evidenced a trend toward superior IL-12 release compared with whole cells. The results showed that bothwhole cells and digested Bifidobacterium sp. are effective at stimulating RAW 264.7 cells to induce the production of nitricoxide and cytokines. The pepsin-pancreatin system used in the current study may be useful in unraveling the mechanism bywhich ingested lactic acid bacteria modulate the induction of macrophage mediators at the cellular level.Key words: macrophage, nitric oxide, cytokine, bifidobacteria

Effects of Bifidobacterium Strains Treated with Gastrointestinal Enzymes on Cytokine Induction in RAW 264.7 Macrophage Cells