Application of bacteroides cellulosilyticus in prevention and/or treatment of inflammatory bowel diseases

A technology of inflammatory bowel disease and cellulose, applied in the field of microorganisms, can solve the problems that the application has not been disclosed, and achieve the effect of great application value, no toxic side effects, and excellent resistance

Pending Publication Date: 2021-10-22
GUANGZHOU ZHIYI PHARMA INC
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Problems solved by technology

Although the bacterium itself and its use in food have been published,...
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Abstract

The invention relates to application of bacteroides cellulosilyticus in prevention and/or treatment of inflammatory bowel diseases, in particular to application of bacteroides cellulosilyticus in preparing medicines, pharmaceutical compositions, food, health-care products, food additives and the like for preventing and/or treating the inflammatory bowel diseases. Results of related in-vivo and in-vitro experiments prove that the bacteroides cellulosilyticus has excellent resistance to inflammatory bowel diseases, including ulcerative colitis and Crohn's disease, and has no toxic or side effect; and the bacteroides cellulosilyticus has durable and effective effect when applied to preparation of the medicines, the pharmaceutical compositions, the food, the health care products or the food additives and the like for preventing and/or treating the inflammatory bowel diseases. The medicines, the pharmaceutical compositions, the food, the health care products or the food additives can be used for preventing and treating the inflammatory bowel diseases and have great application value.

Application Domain

Bacteria material medical ingredientsDigestive system +1

Technology Topic

Crohn's diseaseBowels diseases +10

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  • Application of bacteroides cellulosilyticus in prevention and/or treatment of inflammatory bowel diseases
  • Application of bacteroides cellulosilyticus in prevention and/or treatment of inflammatory bowel diseases
  • Application of bacteroides cellulosilyticus in prevention and/or treatment of inflammatory bowel diseases

Examples

  • Experimental program(6)

Example Embodiment

[0054] Examples of the culture of a decomposer
[0055] Ultrafenal activated strains: DSM 14838T pyraculin propanmetic bacteria, the linear activation is carried out with BHA (cerebral intervention agar medium) plate. Anaerobic, 37 ° C constant temperature culture for 2-7 days, resulting in single colonies.
[0056] Follow Characteristics: After culturing 48 h on BHA plate, it exhibits circular micro-convex, translucent, white, and smooth colonies.
[0057] Microscope Morphology: Cellolatin Pylorgi Performing a Gram Dyeing Mirror, which is a typical rod shape, which is typical rod shape, alone, and in the middle of the middle. figure 1.
[0058] Preparation of the mud: Choose a single colonies in the pancreatic bleak soup for fermentation culture for 8 hours (temperature at a temperature of 37 ° C), the resulting bacteria centrifugally, the rotational speed is 3000r / min, centrifuged for 15 min, to the supernatant, collect the precipitate, Polyfiberus pincobacterial mud.

Example Embodiment

[0059] Example of the drug effect experiment of diphonitrobenzene propagapply bacteria to treat trimitrobenzenesulfonic acid (TNBs) combined with ethanol induced mouse Croan disease
[0060] experimental design
[0061] 60 SPF-class healthy BALB / C mice, females, an average weight of 18 to 22 g, with average weekly age of 6-8 weeks. Adaptive feeding 1 week, randomly groups were as follows: blank group, model group, methazine group, pyraculin propagonal (BC) low, medium and high dose groups.
[0062] Model: In addition to the blank group, once a week, 3% hydrated chloroformolydehyde is injected with intraperitoneal injection. After anesthesia, 2.0 mg TNBS / 50% ethanol is administered 0.1 mL. The blank group was once a week, and the enema was given a bihydrate 0.1 mL. Media for 6 weeks.
[0063] Divided: After 6 weeks of modeling -1 , Low dose 10 6 CFU / only, medium dose 10 8 CFU / only, high dose 10 10 CFU / only. For 14 days, once a day. During modeling and administration, the mice were observed daily, eating drinking water, hair gloss, mental state and activity, and mouse DAI scores were performed according to the standards established by GANTA.
[0064] After 14 days of administration, mice were sacrificed at the 15th day, anatomy, and extracted colon tissue, and cytokine detection was performed.
[0065] Experimental results:
[0066] 1, mice eat water, hair gloss, mental state and activity
[0067] Taking the blank group eating water, hair gloss, mental state and activity as full score (100), score the model group and various administration groups. Such as figure 2 Indicated.
[0068] The model group of eating water, hair gloss, mental state, and activity are significantly lower than the blank group, and the modeling is successful. Although hair gloss is not significantly improved, the positive pharmaceutical beauty sarice is significantly improved by mice eating water, mental state and activity. The effect of betae proponglobacteria is similar to methazine and does not have dose dependence.
[0069] 2, mouse DAI score, the score base is as follows:
[0070] Table 1 DAI rating standard
[0071]
[0072] DAI = body weight loss rate score + stool suspensioned blood / meat blood score, 0-8 points.
[0073] Such as image 3 Indicated.
[0074] The model group scores a long white group (8/0 points), methazine improves blood and rare (5 points), and the score of extrapolatin propoblastine is similar to the methazine, and the cellulin proponorus There is no dose-dependent. It can be seen that the pharmaceutical effects of cellulose propactobacillus alleviate Crohn disease mice are similar to methazine.
[0075] 3, cytokine detection:
[0076] The level of IL-17 and INF-γ cytokines in BALB / C rats were detected by Luminex technology. Depend on Figure 4A , 4B It can be seen that the model group IL-17, INF-γ is much higher than the blank group, and the positive pharmaceutical harazine reduces the level of pro-inflammatory factors; the low, medium and high dosages can also make mice proactive factors in the low, high dosage The level is lowered, which is slightly better than the methazine, but does not have significant. It is illustrated that the cellulose propactor can reduce the acute factor level and have the effect of inhibiting inflammation.
[0077] In summary, the decomposer pylorga has a certain therapeutic effect on Crohn disease, which is similar to methazine and does not have dose dependence.

Example Embodiment

[0078] Examples of Three-Pincellulipin Pikacillus Treatment of Sulfate Sodium Salt Salt (DSS) induced by Chronic Ulcerative Colitis in Rats
[0079] experimental design:
[0080] 60 healthy mature SD rats, male and female, with a body mass 200 ± 20g, purchased from the Guangdong Experimental Animal Center, randomly divided into 6 groups, 10 each group, all rats Adapt to the environment 1 week, feeding ordinary rat feed.
[0081] Solution of 5% sulfate sodium Sodium, DSS, purchased from Sigma, USA, induced by chronic ulcerative colitis, and 60 SD rats were randomly divided into the following groups:
[0082] Normal control group, model group, methazine group, BC low dose group, BC dose group, BC high dose group, using DAI integration and tissue damage score to detect the efficacy of each intervention group. The animal packet table is as follows:
[0083] Table 3 Animal Group Table
[0084] serial number Mold Group Animal only A no Normal control group 10 B Yes Model group 10 C Yes Merazine group 10 D Yes BC low dose group 10 E Yes Distinguishing group in BC 10 F Yes BC high dose group 10
[0085] 1, experimental animal model:
[0086] 50GDSS was added to 1000 ml of distilled water, fully dissolved, and equipped into 5% DSS solution, freshly prepared daily. Method according to Cooper et al (Cooper, HS, Murthy, SN, Shah, RS, Sedergran, DJ, 1993.Clinic opathologic study of dextran sulfate sodium experimental murinecolitis.Laboratory Investigation 69,238-249.), 5% DSS in drinking rats After 7 days of solution, normal drinking water was 10 days, and the above was 1 cycle, repeated four cycles, and the chronic ulcerative colitis model of rats was established. 10 SD rats were a normal control group, and the remaining 50 rats were divided into 6 groups using randomized methods, 10 in each group, respectively, model group, methazine group, BC low dose group, BC Dosage group, BC high dose group, dose design: BC low dose (10 6 CFU / only), dose in BC (10 8 CFU / only), BC high dose (10 10 CFU / only); sarained, 0.4g / kg.d -1 The model group only does not administer the mold; after the mold is successful, it will be administered, once a day, 1 week, continuously intravascation for 1 week.
[0087] 2. Calculation of the activity points of rat disease:
[0088] Hamamoto reference standards (Murano M, MaemuraK, HirataI, ToshinaK, NishikawaT, Hamamoto N, Sasaki S, Saitoh O, Katsu K.The rapeutic effect of intracolonicallyadministered nuclear factor kappa B (p65) antisense oligonucleotide on mousedextran sulphate sodium (DSS) - InducedColitis.Clin Exp Immunol 2000; 120: 51-58), daily observation of the body quality, stool and vulneous blood of rats, calculating the degree of disease integral activity (DAI) per rats, and assessing the degree of colitis activity.
[0089] 3, pathological observation:
[0090] Pre-cooled physiological saline will wash the large intestine, cut 0.5 cm of large intestine at 1 cm from the end of the large intestine, 4% polymethyl formaldehyde soaked, paraffin embed, slice, He staining to do pathological examination. The remainder of colon tissue conducts histopathological score. The degree of histological damage is expressed in inflammation, depth, crypt damage and lesion score score, and take an average as a colon tissue damage count.
[0091] Experimental results
[0092] 1. Experimental observation indicators:
[0093] After successful modeling, the dosage, BC low dose, dose, BC high dose, treatment 1 week, no rat died during the entire experiment, the observation is as follows:
[0094](1) The model group is lowered, the hair is dry, the arch back and the tail, is rare, and some peral can be seen.
[0095] (2) The hair color of the macharazine group is smoother, the spirit is not active, and the reaction is not flexible, but it is better than the model group;
[0096] (3) BC low, medium, high-dose group rat hair color is smoother, the spirit is not active, the reaction is not flexible, similar to the methazine group.
[0097] The above results show: BC low, medium and high dose groups improve the effect of chronic ulcerative colitis and the methazine group.
[0098] 2. Changes in rat disease activities Reference Hamamoto et al. (Murano M, Maemura K, Hiratai, Toshina K, Nishikawa T, Hamamoto N, Sasaki S, Saitoh O, Katsu K.The RapeutiRer Fact of intRacolonically Administered Nuclear Factor Kappa B (P65) AntiSenseoligonucleotide on Mouse Dextran Sulphate Sodium (DSS) -induced colitis.Clinexp Immunol 2000; 120: 51-58) Comprehensive rating of the body quality decreases, stool traits and fecalvascular blood conditions per rat. The results are as follows:
[0099] (1) The normal control group DAI score is always stable in zero level;
[0100] (2) The model group rat inflammation is gradually increased;
[0101] (3) The DAI rating of the methazine group and the bc dose group and the joint group also gradually increased, but it was significantly lower than the model group, see Figure 5.
[0102] The above results show: BC is low, medium and high-dose group comprehensive scores are significantly better than the model group, similar to the methazine group. This suggests that the BC dose group improves the weight loss of chronic ulcerative colitis and the degree of blood and the degree of blood in the blood.
[0103] 3, damage of intestinal mucosa tissue pathological changes like reference Dieleman score (DielemanL A, PalmenMJ, AkolH, etal.Chronic experimental colitis induced by dextran sulfatesodium (DSS) is characterized by Th1 and Th2 Cytokines [J] .Clinical & ExperimentalImmunology, 1999,114 (3): 385-91.)
[0104] Tissue damage score for colon pathological sections per rats. See Image 6 Specifically, the colon inflammation score, lesion score, hood damage, and overall tissue score is divided into various groups of colon inflammation. Normal control group was tested into 0 points, and the model group score 9.40 points. The dose group in the methazine group, BC low dose group, BC, BC high-dose group, tissue damage score is 6.89, 6.65, 7.35, 7.47, and the difference between the dose group and the model group of BC (P <0.01), low dose The effect is slightly better than the methazine, but this difference is not significant.
[0105] The above results show that the pharmacodynamic effect of each dose group in BC is substantially similar to positive pharmaceutical saladidine, which has a significant role in the treatment of chronic ulcerative colitis. This treatment is or derived from the immune function of the extinctellifier propactory, promoting the immune recovery of the columns, thereby effectively correcting inflammatory and anti-inflammatory dynamic imbalance, reducing inflammatory response, reducing the release of inflammatory factors. ability.

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