CN116832070A - Application of saccharomycetes curvularia in preparing medicines for treating and/or improving non-alcoholic fatty liver disease - Google Patents
Application of saccharomycetes curvularia in preparing medicines for treating and/or improving non-alcoholic fatty liver disease Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
Abstract
The invention discloses an application of a sacculus tectorial membrane spore yeast (Saccharomycopsis fibuligera) in preparing a medicine for treating and/or improving non-alcoholic fatty liver disease. The invention also discloses application of the saccharium notatum in preparing medicines for treating and/or improving enteritis and liver lipid accumulation. The invention also discloses application of the saccharum complex film spore yeast in preparing medicines for reducing concentration of glutamic pyruvic transaminase, serum aspartate transaminase, serum triglyceride or serum inflammatory cytokines in serum. The invention proves that the sacculus tectorial membrane spore yeast has obvious improving effect on the non-alcoholic fatty liver; the invention can be applied to the storage of the probiotic preparation for treating the non-alcoholic fatty liver, and has potential application value in the aspect of clinical prevention and treatment of the non-alcoholic fatty liver.
Description
Technical Field
The invention relates to application of a covered cyst membrane spore yeast in preparing a medicine for treating and/or improving non-alcoholic fatty liver disease, belonging to the field of probiotic treatment.
Background
The food and agricultural and world health organizations of the united nations define probiotics as "active microorganisms that bring health benefits to the host when ingested in sufficient quantities", most of which are bacteria similar to the beneficial bacteria naturally found in the human intestinal tract. As a functional food, probiotics have been proved to be capable of improving metabolic diseases such as obesity, diabetes, inflammatory bowel disease and the like in both in vivo and in vitro experiments and clinical application.
Non-alcoholic fatty liver disease (NAFLD), which is a metabolic disease, has been estimated to have a global prevalence of about 25% and has become the most common chronic liver disease. Importantly, sustained fatty liver can denature simple fat into steatohepatitis
(NASH), characterized by the simultaneous presence of inflammation and hepatocellular injury, followed by progression to fibrosis, and ultimately to cirrhosis and hepatocellular carcinoma (HCC). The pathogenesis of non-alcoholic fatty liver is complex, and despite extensive research and development efforts, there is no approved drug specifically directed to metabolic liver disease, and compounds in the later stages of development are generally limited in therapeutic efficacy.
Currently, NAFLD clinically treats multiple targeting FXR, pparα etc. signal pathway targets and inflammatory corpuscles. The propargyl cysteine as described in the patent application (application number: 201910111434.1, propargyl cysteine and the application thereof in preparing new target medicaments) can directly bind to and stimulate the activation of target Epidermal Growth Factor Receptor (EGFR), thereby reducing the blood fat of NAFLD and liver injury caused by NAFLD; the propargyl cysteine described in the patent application can be used for preparing medicines for preventing and treating non-alcoholic fatty liver and non-alcoholic steatohepatitis. However, the targets in this patent application are not novel enough and there may be potential toxic side effects to the targets. Therefore, NAFLD is in urgent need for a new therapeutic drug with little side effects, whereby many scientists direct their eyes to the intestinal flora of the recent strong fire. However, most of the useful treatments for NAFLD are probiotic bacteria, fresh probiotic fungi.
Disclosure of Invention
The invention aims to: the invention aims to solve the technical problem of providing application of the saccharomycete aschersonia (Saccharomycopsis fibuligera) in preparing medicaments for treating and/or improving non-alcoholic fatty liver.
The technical scheme is as follows: in order to solve the technical problems, the invention provides application of the aschersonia aleyrodis (Saccharomycopsis fibuligera) in preparing medicines for treating and/or improving non-alcoholic fatty liver.
The invention also provides application of the saccharomycete aschersonia (Saccharomycopsis fibuligera) in preparing medicines for treating and/or improving enteritis.
The invention also provides application of the saccharomycete aschersonia aleyrodis (Saccharomycopsis fibuligera) in preparing medicaments for treating and/or improving liver lipid accumulation.
The invention also provides application of the saccharomycete aschersonia (Saccharomycopsis fibuligera) in preparing medicaments for reducing the concentration of glutamic pyruvic transaminase in serum.
The invention also provides application of the saccharomycetes curvularia (Saccharomycopsis fibuligera) in preparing medicaments for reducing the concentration of serum aspartate aminotransferase.
The invention also provides application of the aschersonia aleyrodis (Saccharomycopsis fibuligera) in preparing medicines for reducing serum triglyceride concentration.
The invention also provides application of the aschersonia aleyrodis (Saccharomycopsis fibuligera) in preparing medicines for reducing the concentration of serum inflammatory cytokines.
Wherein the concentration of the aschersonia aleyrodis (Saccharomycopsis fibuligera) is 10 8 cfu/ml or more.
Wherein, the dosage form of the medicine comprises tablets, capsules, powder, granules or oral administration agents.
Wherein the medicament comprises a probiotic formulation.
The mechanism of the invention: the Zostera Marinae has the advantages that the weight and liver weight of NAFLD disease mice are obviously improved, the systemic inflammation (spleen weight reduction, IL-6 and IL-1beta reduction) is reduced, the intestinal inflammation caused by NAFLD 'intestinal-liver axis' is reduced (colon shortening is improved), the cell fat in the liver is reduced, and the related NAFLD biochemical indexes of glutamic pyruvic transaminase, aspartate transaminase and triglyceride are obviously reduced in serum.
The beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages: 1. the method has the advantages that the saccharomycete for covering the membrane has obvious improvement effect on the non-alcoholic fatty liver; 2. the invention can be applied to the storage of probiotic preparations for the treatment of non-alcoholic fatty liver, and has potential application value in the aspect of clinical prevention and treatment of non-alcoholic fatty liver.
Drawings
FIG. 1 is a schematic diagram of a mouse experimental design;
FIG. 2 shows the change in body weight of mice in each group;
FIG. 3A is a representative image of the spleens of each group of mice, and FIG. 3B is a statistical plot of the spleens of each group of mice;
fig. 4A is a representative image of the liver of each group of mice, and fig. 4B is a statistical graph of the liver weights of each group of mice;
fig. 5A is a representative image of the colon of each group of mice, and fig. 5B is a statistical plot of the colon length of each group of mice;
FIG. 6A is a representative image of H & E staining of liver sections of mice in each group, and FIG. 6B is a NAS score for the extent of pathology of liver NAFLD in each group;
FIG. 7A is a representative image of H & E staining of the colon of each group of mice, and FIG. 7B is a colon pathology score of each group of mice;
FIG. 8 shows serum alanine aminotransferase levels in mice of each group;
FIG. 9 is serum aspartate aminotransferase levels for each group of mice;
FIG. 10 is serum triglyceride levels for each group of mice;
FIG. 11 shows serum IL-1β levels in mice of each group;
FIG. 12 shows serum IL-6 levels in mice of each group.
Detailed Description
The technical scheme of the invention is further described below with reference to the accompanying drawings.
EXAMPLE 1 cultivation of Saccharomyces curcas and preparation of gastric lavage stock solution for mice
1) Saccharomycetes curcas strain (Saccharomycopsis fibuligera, purchased from Northnano-organism https:// www.bncc.com/, accession number: BNCC 189428) was aerobically cultured in sterilized YM medium (available from Haibo Biotechnology Co., ltd.) at 28℃in a shaker according to the instructions.
2) After 24 hours, the medium was discarded after centrifugation at 500g for 2min at 4℃and the colonies were washed twice with sterilized 1 XPBS (pH 7.4), centrifuged, resuspended in PBS and counted by a cell counter plate using a cell counter, and the total yeast count in PBS was adjusted to 10 8 cfu/ml. These procedures were performed entirely in an ultra clean bench and finally sub-packaged with sterile EP tubes as mouse gastric lavage stock.
EXAMPLE 2 study of Fusarium on the treatment of non-alcoholic fatty liver disease in HFD-fed mice
1) Raising mice: healthy male C57BL/6 mice (from Nanjing university model animals) 5-6 weeks old were housed in a conventional and pathogen-free facility at Nanjing university medical college. All mice were SPF-rated, and the feeding conditions were 23+ -2deg.C and 55+ -5% humidity. The normal drinking of the mice is ensured by adopting 12-hour light/dark alternation. The mice diet and study met the requirements of various aspects of ARRIVE guideline list 2.0. Mice were purchased and fed with maintenance feed for one week for the experiment. The experimental method follows the relevant rules of the experimental animal management committee of Jiangsu province. All experimental procedures were approved by the committee for ethics standards for animal experiments at the university of south Beijing university medical school.
2) Grouping: mice were evenly divided into 3 groups (n=6) after adaptive feeding: CON group: feeding with maintenance feed without other treatment; HFD group (model group): feeding with 60% fat-powered high-fat feed (purchased from Jiangsu province collaborative medical bioengineering Limited liability company) with an average of 20g feed/day; as shown in FIG. 1, the third group (HFD+SF) had the same diet as the model group, and the above-described bursa was infused every other day from week 7Fusarium PBS suspension (number of Fusarium sacchari is 10) 8 cfu/ml) of 100 μl per lavage. The experiment was continued for 7 weeks, and all mice were weighed every other week. The results are shown in figure 2, whereas HFD-fed mice showed a significant increase in body weight compared to chow-fed normal mice (CON group), while HFD-fed mice with gavage had significantly improved body weight (p=0.0171), statistically different body weights (p<0.05)。
3) Tissue sampling: after 7 weeks, the mice were harvested from their eyeballs and placed in a 1.5mL enzyme-free EP tube for 30min, then centrifuged at 2000rpm for 20min, and 50. Mu.L of serum was split into 200. Mu.L EP tubes and stored at-80 ℃. After blood collection, the mice were euthanized by cervical dislocation. The spleen, liver and colon are precisely dissected, and are subjected to subsequent weighing, photographing, cutting and other treatments, and stored at-80 ℃ for further analysis. 1. Representative images and weight statistics of each group of spleens are shown in fig. 3: as inflammation causes splenomegaly, the HFD group has significantly enlarged spleen and increased weight compared to the CON group; whereas the hfd+sf group improved significantly (p=0.0285), spleen weights were statistically significant (p < 0.05). 2. Representative images and weight statistics for each group of livers are shown in fig. 4: NAFLD causes liver lipid accumulation, resulting in increased liver weight and lightening. Liver weight increased and color lighter in HFD group compared to CON group; the hfd+sf group significantly improved the color of liver weight and fatty liver, the color was significantly darkened, and liver weight was statistically significant (p=0.0026 < 0.05). 3. Representative images and length statistics for each set of colon are shown in fig. 5: NAFLD is often accompanied by enteritis, resulting in abnormal shortening of the colon. Compared with the CON group, the HFD group showed significantly shorter colon length, and the treatment with the aschersonia aleyrodis improved abnormal shortening of the colon (p=0.0027), with a statistically significant colon length (p < 0.05).
4) H & E staining
A portion of the colon and liver of the mice was fixed in 4% paraformaldehyde, embedded in paraffin, sectioned into paraffin sections, then stained with hematoxylin and eosin (H & E), examined under a microscope (Nikon ECLIPSE Ti-U) for tissue morphology and photographed. The results showed representative images of liver section hematoxylin and eosin (H & E) staining and NAS scores reflecting the extent of liver NAFLD lesions (fig. 6): compared with CON group, the HFD group liver cells generate obvious balloon-like and fat changes, and the liver cells are obviously improved after the treatment of the aschersonia aleyrodis; NAS scoring of liver NAFLD lesion extent showed that HFD group liver lesions were severe and significantly improved (p=0.0054) after treatment with aschersonia aleyrodis had statistical significance (p < 0.05). Representative images of colon H & E staining and colon histological scores are shown in fig. 7: compared to the CON group, the HFD group showed a disruption of the intestinal barrier, a pathological change in the colonic histomorphology, a significant improvement in the hfd+sf group after treatment (p= 0.0343), and a statistically significant colonic histological score (p < 0.05).
5) Serum biochemical index detection
The detection was performed on a fully automatic biochemical analyzer (Rayto, chemray 800) according to the instructions using glutamic pyruvic transaminase, aspartic transaminase, triglyceride assay kit (purchased from shenzhen radu life sciences ltd). The results show that: serum glutamic pyruvic transaminase (ALT) was significantly increased in the HFD group compared to the CON group, significantly decreased (p < 0.0001) after treatment in the hfd+sf group, and serum ALT levels were statistically significant (p < 0.05) (fig. 8); the HFD group had a significant increase in serum aspartate Aminotransferase (AST) and serum Triglyceride (TG) compared to the CON group, and the hfd+sf group had a decrease after treatment, but no significant difference (fig. 9-10);
6) Serum ELISA experiments
Serum assays were performed as indicated using the Mouse IL-1beta Uncoated ELISA kit and the Mouse IL-6Uncoated ELISA kit (both available from Simer Feishmania technologies). The results show that: serum IL-1β and IL-6 were significantly elevated in HFD versus SF group compared to CON group, serum IL-1β was significantly reduced (p= 0.0281) after treatment, statistically significant (p < 0.05), and IL-6 was reduced with no statistical difference (fig. 11-12).
Claims (10)
1. Application of Saccharomycetes curcas (Saccharomycopsis fibuligera) in preparing medicaments for treating and/or improving non-alcoholic fatty liver disease.
2. Use of a oocyst membrane-covered yeast (Saccharomycopsis fibuligera) in the manufacture of a medicament for treating and/or ameliorating enteritis.
3. Use of a rhodosporidium cinquefoil (Saccharomycopsis fibuligera) for the preparation of a medicament for the treatment and/or amelioration of liver lipid accumulation.
4. Application of Saccharomycetes curcas (Saccharomycopsis fibuligera) in preparing medicines for reducing glutamic pyruvic transaminase concentration in serum is provided.
5. Use of a rhodosporidium curvatus (Saccharomycopsis fibuligera) for the preparation of a medicament for reducing serum aspartate aminotransferase concentration.
6. Use of a rhodosporidium curvatus (Saccharomycopsis fibuligera) for the preparation of a medicament for reducing serum triglyceride concentration.
7. Use of a rhodosporidium curvulus (Saccharomycopsis fibuligera) for the preparation of a medicament for reducing serum inflammatory cytokine concentration.
8. The use according to any one of claims 1 to 7, wherein the concentration of said aschersonia aleyrodis (Saccharomycopsis fibuligera) is 10 8 cfu/ml or more.
9. The use according to any one of claims 1 to 7, wherein the pharmaceutical dosage form comprises a tablet, capsule, powder, granule or oral dosage form.
10. The use according to any one of claims 1 to 7, wherein the medicament comprises a probiotic preparation.
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| CN202310739132.5A CN116832070A (en) | 2023-06-21 | 2023-06-21 | Application of saccharomycetes curvularia in preparing medicines for treating and/or improving non-alcoholic fatty liver disease |
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| CN202310739132.5A CN116832070A (en) | 2023-06-21 | 2023-06-21 | Application of saccharomycetes curvularia in preparing medicines for treating and/or improving non-alcoholic fatty liver disease |
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|---|---|
| CN (1) | CN116832070A (en) |
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|---|---|---|---|---|
| WO1999041996A1 (en) * | 1998-02-20 | 1999-08-26 | Almqvist & Wallenbeck Ab | Probiotic composition and the use thereof |
| KR101334157B1 (en) * | 2013-03-12 | 2013-11-29 | 농업회사법인 주식회사 엘바이오텍 | Surfur removing poison produced using effective microorganism and uses thereof |
| CN111088174A (en) * | 2019-12-30 | 2020-05-01 | 上海应用技术大学 | Saccharomycetes Fungiensis with oxidation resistance and whitening effect and application thereof |
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2023
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|---|---|---|---|---|
| WO1999041996A1 (en) * | 1998-02-20 | 1999-08-26 | Almqvist & Wallenbeck Ab | Probiotic composition and the use thereof |
| KR101334157B1 (en) * | 2013-03-12 | 2013-11-29 | 농업회사법인 주식회사 엘바이오텍 | Surfur removing poison produced using effective microorganism and uses thereof |
| CN111088174A (en) * | 2019-12-30 | 2020-05-01 | 上海应用技术大学 | Saccharomycetes Fungiensis with oxidation resistance and whitening effect and application thereof |
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| SUI WANG 等: "The important role of fungi in inflammatory bowel diseases", SCANDINAVIAN JOURNAL OF GASTROENTEROLOGY, vol. 56, no. 11, 31 December 2021 (2021-12-31), pages 1312 * |
| ZHIJIE DAN 等: "Effects of fishmeal substitution by four fermented soybean meals on growth， antioxidant capacity and inflammation responses of turbot juveniles (Scophthalmus maximus L.)", AQUACULTURE, 28 May 2022 (2022-05-28), pages 10 * |
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