Lactobacillus separation strains with anti-inflammatory activity and application thereof
An isolated strain, Lactobacillus technology, applied to Lactobacillus isolates with anti-inflammatory activity and the field of use thereof, can solve problems such as reduced resistance to infection and the like
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Embodiment 1
[0071] Embodiment 1. has the preliminary screening experimental material and method of the lactobacillus isolate strain of anti-inflammatory activity:
[0072] A. Source and preparation of test strains:
[0073] The applicant obtained the gastrointestinal tract specimens of many healthy adults through the Affiliated Hospital of China Medical University (Taichung City) in February 1991, and isolated more than 400 strains of Lactobacillus isolates from these specimens. In order to find probiotics with the potential to be used in the treatment of rheumatoid arthritis, applicants analyzed the anti-inflammatory activity of these Lactobacillus isolates and used IL-10 as a screening marker.
[0074] Each test strain was inoculated into Lactobacillus Bacto MRS broth medium (DIFCO, Cat. No. 0881), and then anaerobically cultured at 37° C. for 12 to 18 hours. The resulting bacterial culture was centrifuged at 4,000 rpm for 15 minutes, the supernatant was discarded, and the precipitate ...
Embodiment 2
[0091] Example 2. Evaluation of Lactobacillus isolates stimulating the ability of human PBMC to secrete IL-10 and IFN-γ
[0092] Experimental materials and methods:
[0093] A. Preparation of human peripheral blood mononuclear cells:
[0094] Take human white blood cell concentrate (Tainan Blood Donation Center) and Ficoll-Paque that have passed the inspection and are used for experiments TM PLUS (17-1441-03, Amersham Biosciences) was used for density gradient centrifugation (720 g x 20 min at 4° C.) at a volume ratio of 1:1. Afterwards, the erythrocytes were removed, and the resulting human PBMCs were adjusted to a cell concentration of 4×10 in RPMI1640 medium containing 10% FBS. 6 cells / mL for later use.
[0095] B. Evaluation of Lactobacillus isolates stimulating human PBMC to secrete IL-10:
[0096] The ability of Lactobacillus isolates GMNL-19, GMNL-76, GMNL-78, GMNL-89, GMNL-94, GMNL-101 and GMNL-161 to stimulate human PBMC to secrete IL-10 is basically the "experim...
Embodiment 3
[0118] Example 3. Evaluation of the therapeutic effect of Lactobacillus isolates GMNL-76 and GMNL-89 on rats with collagen-induced arthritis
[0119] Experimental materials and methods:
[0120] A. Experimental animals:
[0121] Male LEW / SsNNarl rats (6 weeks old, weighing approximately 200 to 250 g) obtained from the National Applied Research Laboratories National Laboratory Animal Center were used in the following experiments. All experimental animals were kept in an individually air-conditioned animal room with 12 hours of light and 12 hours of darkness, room temperature maintained at 25±1°C and humidity maintained at 60±5%, and water and feed were adequately supplied. Give the animals a week to acclimate to the environment, and start the experiment after the animals are stable. The handling of experimental animals and all experimental procedures are carried out in accordance with the standard regulations of the Experimental Animal Committee.
[0122] B. Preparation of t...
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