| Summary: | 碩士 === 國立中興大學 === 動物科學系所 === 107 === Lactobacillus paracasei Y310 has been proved to be able to produce anti-inflammatory tryptophan metabolite. Lb. paracasei shows anti-inflammatory ability both in vitro and in vivo, including inhibition of lipopolysaccharide (LPS)-induced pro-inflammatory cytokine production in murine macrophage cell line, improvement of chemical-induced colitis in mice, as well as inhibition of intraperitoneal LPS infection-induced inflammation in Lanyu pig model. However, considering the fermentation cost, using alternative culture medium is more economic for commercialization. Accordingly, the aim of this study was to use two kinds of whey-derived materials, including whey protein concentrate (WPC) and lactose as the alternative medium materials for cultivation of Lb. paracasei Y310, and further evaluated the anti-inflammatory ability in vitro and in dextran sulfate sodium (DSS)-induced mice colitis prevention model.
Firstly, we selected and adjusted alternative media materials. Since Lb. paracasei Y310 was isolated from raw milk, we compared the effects of different types of milk-derived materials on the growth performance and anti-inflammatory tryptophan metabolite production of Lb. paracasei Y310. The result revealed that using whey powder as the medium could produce more tryptophan metabolite than which in milk powder and skim milk powder. According to the result, we chose whey powder as the major material for culturing Lb. paracasei Y310. However, the casein residue in whey powder coagulated after fermentation. Therefore, we further change WPC as nitrogen source and lactose as carbon source for formula adjustment. The results showed that the use of 40.4% WPC and 47.92% lactose did not cause coagulation, and the number of bacteria remained above 108 CFU/mL. In addition, the anti-inflammatory performance of Lb. paracasei Y310 cultivated by WPC and lactose alternative medium in mice microphage RAW 264.7 cell line and DSS-induced mice colitis prevention model were evaluated. The in vitro trial used Lb. paracasei Y310 cultivated by WPC and lactose alternative medium or commercial culture medium MRS broth (104, 105, 106 CFU/mL) for microphage co-culture, after that we collected the supernatant for cytokine analysis. The result revealed that, Lb. paracasei Y310 could induce cytokine productions in murine macrophage cells. Furthermore, in the mice colitis prevention model, 8 weeks old C57BL/6 female mice were used for a total of 14 days in the experimental period. During the period, each mouse was orally administered with 107 CFU or 108 CFU of Lb. paracasei Y310 cultivated by WPC and lactose alternative medium or commercial MRS broth respectively. The result revealed that, whether using WPC and lactose alternative medium or MRS broth, oral administration of 107 CFU and 108 CFU Lb. paracasei Y310 improved the symptoms in mice. But in histopathological evaluation, only administration of Lb. paracasei Y310 that cultivated by MRS broth could improve the damage induced by DSS.
In summary, the use of WPC and lactose may have the potential as alternative medium for anti-inflammatory probiotic Lb. paracasei Y310 to reduce the cost of fermentation. In further studies, the difference between the production of anti-inflammatory tryptophan metabolites in alternative medium, and the interaction between metabolites and anti-inflammatory tissues still needs to be investigated.
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