Pediococcus acidilactici CCEM6432 capable of reliving depression, fermentation food thereof and application thereof
A technology of CCFM6432 and CCFM64322, applied in the field of Pediococcus lactis CCFM6432 and its fermented food, can solve the problems of large side effects, slow onset of action, inconsistent treatment effect, etc., to increase biosynthesis, increase abundance, and restore intestinal flora disorder. Effect
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Embodiment 1
[0062] Example 1: Pediococcus lactis CCFM6432 can reduce depression-like behavior in mice
[0063] Thirty-two 6-week-old male C57BL / 6J mice were randomly divided into four groups according to body weight after acclimating to the environment for one week: normal control group, depression model group, drug intervention group, and CCFM6432 intervention group, with 8 mice in each group. The animal groups and treatment methods are shown in Table 1.
[0064] Table 1 Grouping and processing methods of animal experiments
[0065]
[0066] Chronic unpredictable stress depression mouse model: 1-2 kinds of stimuli are randomly used every day, and the time of stimulation is randomly determined every day to avoid circadian rhythm. No more than three times of each method for a period of five weeks. Stimulus factors include: (1) fasting for 24 hours; (2) water deprivation + empty bottle stimulation for 24 hours; (3) tail clamping for 3 minutes; (4) wet litter for 24 hours; (5) braking f...
Embodiment 2
[0075] Example 2: CCFM6432 can significantly increase the level of neurotransmitter (5-HT) in the brain of depressed mice
[0076] The mice in Example 2 were euthanized at the end of the sixth week, and the brain tissues of the mice were taken, and the brainstem and prefrontal cortex were separated on ice. Take a certain amount of fresh brainstem and prefrontal cortex tissue (weight not less than 50mg), add 9 times the volume of sterile PBS buffer (equivalent to 1g tissue plus 9mL homogenate), and use a tissue homogenizer to homogenize Slurry and tissue fluid were centrifuged at 3000g for 20min to take the supernatant, added an equal volume of 5% perchloric acid to precipitate protein, centrifuged at 10000g for 10min, absorbed the supernatant and filtered it through a 0.22μm water filter membrane, and detected 5-HT with an ELISA kit content. Experimental results such as image 3 As shown, the results showed that administration of CCFM6432 could significantly reverse the redu...
Embodiment 3
[0077] Example 3: Pediococcus lactis CCFM6432 can stimulate enterochromaffin cells to secrete 5-HTP
[0078] Determination of 5-HTP in cell supernatant: RIN14B cells in 4×10 5 / mL density in 24-well plate, incubated for 72h. Discard the medium, wash the cells with HBSS (1 mL) containing 0.1% bovine serum albumin (BSA) and 2 μM fluoxetine, add 1 mL of HBSS suspension containing CCFM6432 (the control group is HBSS without bacteria), 37 Incubate at ℃ for 20 minutes, collect the supernatant, centrifuge at 6000g for 5 minutes to remove the precipitate, and take the supernatant and freeze it at -80℃ for testing. 5-HTP in the cell supernatant was detected by HPLC-FLD (reference example 3).
[0079] Tryptophan hydroxylase 1 (TPH1) mRNA determination: the adherent cells in the above steps were washed three times with HBSS, added 1mL Trizol, incubated on ice for 5-10min, blown to detach the cells, and transferred the lysate to a dry place. Enzyme EP tube. Total cellular RNA was extr...
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