Probiotic preparation for preventing and treating vascular dementia and preparation method of probiotic preparation
A technology of vascular dementia and strains, applied in the field of biopharmaceuticals, can solve problems such as unreported, and achieve the effect of convenient use, good ecological mutual assistance, and improvement of intestinal flora imbalance
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Embodiment 1
[0026] Example 1 Isolation and identification of Clostridium butyricum (Clostridium butyricum) WZMC1016 bacterial strain
[0027] Using the feces of a healthy young man as an isolated sample, on the improved MRS agar medium, under anaerobic conditions at 37°C, smear, isolate and cultivate for 48 hours to obtain Clostridium butyricum WZMC1016 according to the present invention, and identify Clostridium butyricum.
[0028] The formula of the improved MRS medium: peptone 10g, beef extract 10g, yeast extract 5g, glucose 20g, K 2 HPO 4 2g, MgSO 4 7 hours 2 O 0.5g, MnSO 4 4H 2 O 0.2g, fructooligosaccharide 3g, diammonium citrate 2g, agar 15g, distilled water 1L, adjust the pH to 7.0, and sterilize at 115°C for 15 minutes.
[0029] Clostridium butyricum (Clostridium butyricum) WZMC1016 of the present invention has the following microbiological characteristics:
[0030] (1) Colony shape: milky white, round and slightly convex, with irregular edges, 1-3 mm in diameter, slightly...
Embodiment 2
[0035] Preparation of Example 2 Clostridium butyricum (Clostridium butyricum) WZMC1016 probiotic preparation composition
[0036] 1. Seed solution preparation: pick a single colony of Clostridium butyricum (Clostridium butyricum) WZMC1016, inoculate it in 50 mL of modified MRS liquid medium, pH 7.0±0.5, 37°C, anaerobic culture for 24 hours, that is the seed solution;
[0037] The formula of the improved MRS medium: peptone 10g, beef extract 10g, yeast extract 5g, glucose 20g, sodium acetate 5g, K 2 HPO 4 2g, MgSO 4 7 hours 2 O 0.5g, MnSO 4 4H 2 O 0.2g, fructooligosaccharide 3g, diammonium citrate 2g, distilled water 1L, adjust the pH to 7.0, and sterilize at 115°C for 15min.
[0038] 2. Fermentation culture: Take the seed solution in step 1, inoculate it into a new improved MRS medium with an inoculum amount of 1%, culture it anaerobically at 37°C for 24 hours, and end the culture;
[0039] 3. Collect the bacteria: take the fermentation liquid in step 2, remove the ferm...
Embodiment 3
[0042] Therapeutic animal experiment of embodiment 3 vascular dementia
[0043] Objective: To observe the effect of the Clostridium butyricum preparation of the present invention on the learning and memory, the oxidation index in the brain and the morphological changes of nerve cells in mice with vascular dementia.
[0044] 1. Materials and methods:
[0045] 1. Drug: Clostridium butyricum (Clostridium butyricum) WZMC1016 bacteria powder, the number of viable bacteria is 1×10 8 CFU / g, the specific preparation method, see Example 2 for details.
[0046] 2. Experimental animals: male ICR mice, weighing 18-22 grams.
[0047] 3. Vascular dementia (VaD) mouse model preparation and animal grouping: permanent unilateral right common carotid artery occlusion (rUCCAO), mice were anesthetized, fixed on the operating table, separated and sutured with permanent small silk thread The right common carotid artery was performed, and the body temperature of the mice was kept at a constant te...
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