CN111117935A - Microbial agent for inhibiting muscle synthesis and application thereof - Google Patents
Microbial agent for inhibiting muscle synthesis and application thereof Download PDFInfo
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- CN111117935A CN111117935A CN202010084590.6A CN202010084590A CN111117935A CN 111117935 A CN111117935 A CN 111117935A CN 202010084590 A CN202010084590 A CN 202010084590A CN 111117935 A CN111117935 A CN 111117935A
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- 210000003205 muscle Anatomy 0.000 title claims abstract description 29
- 230000000813 microbial effect Effects 0.000 title claims abstract description 19
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 17
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 17
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 15
- 241000194017 Streptococcus Species 0.000 claims abstract description 27
- 239000003795 chemical substances by application Substances 0.000 claims description 14
- 239000003814 drug Substances 0.000 claims description 11
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims 1
- 201000000585 muscular atrophy Diseases 0.000 abstract description 30
- 206010028289 Muscle atrophy Diseases 0.000 abstract description 29
- 230000020763 muscle atrophy Effects 0.000 abstract description 28
- 230000000968 intestinal effect Effects 0.000 abstract description 18
- 244000005700 microbiome Species 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 3
- 208000024891 symptom Diseases 0.000 abstract description 2
- 230000006872 improvement Effects 0.000 description 16
- 108090000623 proteins and genes Proteins 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 235000021120 animal protein Nutrition 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 241000606125 Bacteroides Species 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- 241000194033 Enterococcus Species 0.000 description 4
- 241000589565 Flavobacterium Species 0.000 description 4
- 241000588748 Klebsiella Species 0.000 description 4
- 241001057811 Paracoccus <mealybug> Species 0.000 description 4
- 241000192031 Ruminococcus Species 0.000 description 4
- 241001148134 Veillonella Species 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000007774 longterm Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 238000002591 computed tomography Methods 0.000 description 3
- 230000000875 corresponding effect Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 241000193403 Clostridium Species 0.000 description 2
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 2
- 241000605909 Fusobacterium Species 0.000 description 2
- 108010064851 Plant Proteins Proteins 0.000 description 2
- 241000605947 Roseburia Species 0.000 description 2
- 241001608455 Rossia <Bacillariophyta> Species 0.000 description 2
- 108010046377 Whey Proteins Proteins 0.000 description 2
- 102000007544 Whey Proteins Human genes 0.000 description 2
- 235000019577 caloric intake Nutrition 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 238000010219 correlation analysis Methods 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 235000021118 plant-derived protein Nutrition 0.000 description 2
- 235000021075 protein intake Nutrition 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 229940071440 soy protein isolate Drugs 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 235000021119 whey protein Nutrition 0.000 description 2
- 108091093088 Amplicon Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
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- 230000001684 chronic effect Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 235000021196 dietary intervention Nutrition 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
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- 230000036541 health Effects 0.000 description 1
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- 238000011134 hematopoietic stem cell transplantation Methods 0.000 description 1
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- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000025161 regulation of muscle atrophy Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 210000001578 tight junction Anatomy 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
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- Bioinformatics & Cheminformatics (AREA)
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Abstract
The invention provides a microbial agent for inhibiting muscle synthesis and application thereof, relating to the technical field of application of microorganisms; the microbial agent comprises Streptococcus enterica (Streptococcus). The intestinal flora in the microbial agent can possibly cause muscle atrophy or promote the development of the muscle atrophy, and can be used as a new direction for treating the symptoms of the muscle atrophy.
Description
Technical Field
The invention belongs to the technical field of application of microorganisms, and particularly relates to a microbial agent for inhibiting muscle synthesis and application thereof.
Background
Skeletal muscle accounts for about 40% of body weight and plays a vital role in maintaining human health, including amino acid and glucose metabolism, protein storage and exercise. Muscle atrophy is a chronic metabolic disease commonly faced by people in modern society, and people lack of exercise for a long time, old people, cancer patients and people treated by large dose of hormones face different degrees of muscle atrophy. Studies have shown that muscle atrophy has become an independent factor affecting death. Therefore, how to effectively improve muscle atrophy has become a health problem to be solved urgently.
The previous research is mainly to research the mechanism of muscle atrophy from the perspective of cell signaling pathways. Unfortunately, it has not been possible to screen for drugs that either block the cell signaling pathways associated with regulation of muscle atrophy, or that do not produce new side effects. Therefore, new ways for improvement are urgently sought. Since the human intestinal flora is a complex ecosystem, it can influence the absorption of nutrients by biotransformation of food complexes, while their metabolites can enter the blood circulation through the intestinal tight junctions and enter the muscle tissue. These research clues suggest that intestinal flora may be an effective target for regulating muscle atrophy. However, no research is available to find intestinal microbial strains capable of promoting muscle atrophy and inhibiting muscle synthesis.
Disclosure of Invention
In view of the above, the present invention aims to provide a microbial agent for inhibiting muscle synthesis, which can promote muscle atrophy and inhibit muscle synthesis, and has a significant reverse relationship with respect to improvement of muscle atrophy, thereby providing a new basis for deep-level mechanism research of muscle atrophy.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a microbial agent for inhibiting muscle synthesis, which comprises Streptococcus enteric (Streptococcus).
The invention also provides a medicament for inhibiting muscle synthesis, and the active ingredient of the medicament comprises the microbial agent.
Preferably, the medicine also comprises pharmaceutically acceptable auxiliary materials.
The invention provides a microbial agent for inhibiting muscle synthesis, which comprises Streptococcus enteric (Streptococcus). The intestinal flora is obtained by screening the excrement of a hematopoietic stem cell transplantation patient with long-term muscle atrophy by a clinical queuing method; then, the abundance value of the intestinal flora is correlated with the improvement degree of corresponding muscle atrophy, and the correlation coefficient of Streptococcus (Streptococcus) to the change of muscle grasping force reaches-0.895 (p is 0.016) and the correlation coefficient of Streptococcus (Streptococcus) to the change of muscle area reaches-0.949 (p is 0.004). The intestinal flora may therefore induce or promote the development of muscle atrophy, as a new direction for the treatment of symptoms of muscle atrophy.
Drawings
FIG. 1 is a flow chart of the screening of intestinal microorganisms;
FIG. 2 is a graph showing the correlation between Streptococcus (Streptococcus) and changes in muscle grip;
FIG. 3 is a graph showing the correlation between Streptococcus (Streptococcus) and changes in muscle area.
Detailed Description
The invention provides a microbial agent for inhibiting muscle synthesis, which comprises Streptococcus enteric (Streptococcus).
The source of the microbial agent is not particularly limited, and the microbial agent can be obtained from commercially available common strains (strains) and can also be screened from intestinal tracts or intestinal excretions. In the present example, for the purpose of illustrating the effects, the intestinal excreta of a hematopoietic stem cell transplant patient with long-term muscle wasting was screened according to the flowchart shown in fig. 1. The present invention preferably further comprises prior to said screening, performing a two month quality protein nutritional intervention on said patient, said intervention preferably comprising caloric intake of 35kcal/kg/d, protein intake of 1.5 g/kg/d; the protein comprises animal protein and plant protein, and the mass ratio of the animal protein to the plant protein is 1: 1. The animal protein of the present invention preferably comprises a soy protein isolate, preferably purchased from the prince royal group (cat No. YP928H, shandong, china). The animal protein according to the present invention preferably comprises whey protein, preferably purchased from Livingston (hilmr 9410, Livingston, CA, USA). According to the invention, the patients after the dry prognosis are preferably judged to have the muscle atrophy improvement effect through muscle CT scanning, and are divided into two groups (the muscle atrophy improvement population and the non-improvement population) according to the quality of the muscle improvement effect, and the intestinal microorganism specimens of the two groups of populations before and after protein intervention are subjected to 16srDNA sequencing respectively. After the sequencing, the sequence set is divided into operable classification units (OTUs) by using UCLUST under a 97% consistency threshold, and each OTU is classified and distributed by similarity through a GLSEARCH program; then using Metastats analysis (analysis of significant difference between groups), which improved the population by comparing muscle atrophy and did not improve the intestinal strain of 1/5 before abundance of intestinal flora in the population, among the strains with significant statistical difference, i.e., Enterococcus (Enterococcus), Veillonella (Veillonella), Ruminococcus (Ruminococcus), Bacteroides (Bacteroides), Klebsiella (Klebsiella), flavobacterium (flavobacterium), clostridium (Fusobacterium), paracoccus (paracoccus), Streptococcus (strepococcus) and rossia (Roseburia) were searched for by the analysis of difference between groups; finally, the differential bacteria are subjected to correlation analysis according to the abundance value and the corresponding muscle atrophy improvement degree, and Streptococcus (Streptococcus) is found to show a remarkable backward correlation with the muscle atrophy improvement and show a remarkable forward correlation with the inhibition of muscle synthesis.
The invention also provides a medicament for inhibiting muscle synthesis, and the active ingredient of the medicament comprises the microbial agent. The medicine of the invention preferably also comprises pharmaceutically acceptable auxiliary materials. The dosage form of the drug is not particularly limited in the present invention.
The following examples are provided to illustrate the microbial agents for inhibiting muscle synthesis of the present invention in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
The protocol of this example was carried out using the method of clinical Cohort (Cohort study). The embodiment of the invention is registered by a Chinese clinical registration center (ChiCTR 1800017765).
Screening was performed according to the scheme shown in FIG. 1: by recruiting hematopoietic stem cell transplant patients with long-term muscular atrophy and by performing a two month long-term intervention with high quality protein nutrition (35 kcal/kg/d caloric intake, 1.5g/kg/d protein intake; wherein the protein consists of animal protein and vegetable protein in a 1:1 mass ratio, wherein the vegetable protein is soy protein isolate (YP928H, limon, shandong, china) and the animal protein is whey protein (hilar 9410, Livingston, CA, USA)) on all cohorts. The muscle atrophy improvement effect was judged by muscle CT scan two months later. Then, dividing the group into two groups according to the quality of the muscle improvement effect (the muscle CT scanning and the holding power value after the intervention of the patient, calculating the difference value of the CT muscle area change and the holding power change before and after the intervention, listing the group with positive difference values of the two indexes as a muscle improvement group, listing the group with negative difference values of the two indexes as a muscle atrophy non-improvement group), and respectively carrying out 16srDNA sequencing on the intestinal microorganism samples before and after the protein intervention of the two groups of people.
The intestinal flora 16srDNA analysis method comprises the following steps:
DNA was extracted from the thawed fecal samples. DNA quality was determined by 2% agarose gel electrophoresis and spectrophotometry. The V3-V4 region of the 16srDNA gene was amplified with Fastpfu DNA polymerase (TransStartTM, TransGen Biotech).
The primer adopted is 336F (SEQ ID NO. 1): 5'-GTACTCCTAGGGGGCAGCA-3', respectively;
806R(SEQ ID NO.2)：5'-GTGGACTACHVGGGTWTCTAAT-3'。
the PCR cycling conditions were as follows: pre-denaturation at 95 ℃ for 5 min; denaturation at 85 ℃ for 45s, annealing at 55 ℃ for 50s, extension at 72 ℃ for 45s, 28 cycles; prolonging the temperature at 72 ℃ for 10 min.
The purified amplicons were pooled in equimolar amounts and paired-end sequencing was performed according to standard protocols on the Illumina MiSeq platform (2 × 300). The sequence set is divided into actionable taxons at a consistency threshold of 97% using UCLUST (otu). Using the GLSEARCH program, each OTU was assigned by similarity, yielding a total of 1311239 high quality sequences with coverage over 99%. The OTU representative sequences are analyzed by alignment using RDP Classifier algorithm (default) or blast, uclust consensus xonomy assigner and the like, and species information of the colonies is annotated at various levels (kingdom, phylum, compendium, family, genus and species).
Method for the study of improvement of Streptococcus (Streptococcus) and muscle atrophy:
using Metastats analysis, i.e., analysis of significant differences between groups, which improved the population by comparison to muscle atrophy and did not improve the intestinal strain of pre-abundance 1/5 of intestinal flora in the population, strains with significant statistical differences were sought by analysis of differences between groups, i.e., Enterococcus (Enterococcus), Veillonella (Veillonella), Ruminococcus (Ruminococcus), Bacteroides (Bacteroides), Klebsiella (Klebsiella), flavobacterium (flavobacterium), clostridium (Fusobacterium), paracoccus (paracoccus), Streptococcus (Streptococcus), and rossia (Roseburia); finally, the differential bacteria are subjected to correlation analysis according to the abundance value and the corresponding muscle atrophy improvement degree (the result is shown in figures 2-3), and the Streptococcus (Streptococcus) is found to be in a significant reverse correlation with the muscle atrophy improvement degree.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
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<120> microbial agent for inhibiting muscle synthesis and application thereof
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Claims (3)
1. A microbial agent for inhibiting muscle synthesis, which comprises Streptococcus enterica (Streptococcus).
2. A medicament for inhibiting muscle synthesis, wherein the active ingredient of the medicament comprises the microbial agent of claim 1.
3. The medicament of claim 2, further comprising a pharmaceutically acceptable excipient.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010084590.6A CN111117935A (en) | 2020-02-10 | 2020-02-10 | Microbial agent for inhibiting muscle synthesis and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010084590.6A CN111117935A (en) | 2020-02-10 | 2020-02-10 | Microbial agent for inhibiting muscle synthesis and application thereof |
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| CN202010084590.6A Pending CN111117935A (en) | 2020-02-10 | 2020-02-10 | Microbial agent for inhibiting muscle synthesis and application thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102686112A (en) * | 2009-10-27 | 2012-09-19 | 塞路尔控股有限公司 | Novel strains of lactic acid bacteria and combinations thereof for the manufacture of probiotic preparations |
| CN109517765A (en) * | 2019-01-11 | 2019-03-26 | 谭瑛 | A kind of streptococcus fecalis and its application |
| CN110461337A (en) * | 2017-02-23 | 2019-11-15 | 英特塞普特医药品公司 | Pharmaceutical compositions with bile acid derivatives and microbiome and uses thereof |
| CN110452860A (en) * | 2019-09-24 | 2019-11-15 | 厦门大学 | A streptococcus salivarius and its application in the treatment of inflammatory bowel disease |
-
2020
- 2020-02-10 CN CN202010084590.6A patent/CN111117935A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102686112A (en) * | 2009-10-27 | 2012-09-19 | 塞路尔控股有限公司 | Novel strains of lactic acid bacteria and combinations thereof for the manufacture of probiotic preparations |
| CN110461337A (en) * | 2017-02-23 | 2019-11-15 | 英特塞普特医药品公司 | Pharmaceutical compositions with bile acid derivatives and microbiome and uses thereof |
| CN109517765A (en) * | 2019-01-11 | 2019-03-26 | 谭瑛 | A kind of streptococcus fecalis and its application |
| CN110452860A (en) * | 2019-09-24 | 2019-11-15 | 厦门大学 | A streptococcus salivarius and its application in the treatment of inflammatory bowel disease |
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