CN112043680A - Application of nano-silver preparation in improvement of low-grade inflammation and intestinal flora disorder related to obesity induced by high-fat diet - Google Patents
Application of nano-silver preparation in improvement of low-grade inflammation and intestinal flora disorder related to obesity induced by high-fat diet Download PDFInfo
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Abstract
The invention discloses a nano-silver preparation for improving obesity-related low-grade inflammation and intestinal flora disorder and application thereof. According to the invention, through the related experiments of the nano-silver exposure on normal and obese mice, the treatment of a mouse model induced by high-fat diet by using the nano-silver preparation is realized for the first time, the lowest concentration of the nano-silver preparation which generates a significant antibacterial effect (p <0.05) on E.coli is less than or equal to 5.0mg/L, the nano-silver preparation does not show a significant growth inhibition effect on S.aureus under the highest test dose (50mg/L), the nano-silver preparation has a regulating effect on intestinal microorganisms of verruca microbia, deironium, Epsilonbacterota and actinomycetemta, and the nano-silver preparation also shows a significant slow inhibition effect on the systemic low-grade inflammation of the mice induced by the high-fat diet.
Description
Technical Field
The invention belongs to the field of nano-biomedicine, and particularly relates to application of a nano-silver preparation in improvement of high-fat diet-induced obesity-related low-grade inflammation and intestinal flora disorder.
Background
With the change of dietary structure and life style of people, obesity has become a global problem. Obesity can significantly increase the risk of death and the risk of developing fatty liver, diabetes, hypertension, insulin resistance syndrome, and even certain cancers in an individual. In recent years, a number of studies have shown that low grade inflammation and intestinal microbial disturbances play a crucial role in the development of obesity and related diseases. However, how to effectively intervene in low-grade inflammation and intestinal microbial disorder related to obesity and further relieve the occurrence process of obesity-related diseases is a scientific problem to be solved urgently in the field of nutrition and health research.
Low grade inflammation (low-grade inflammation), unlike acute inflammation, can produce long-term and systemic effects. Low grade inflammation is often characterized by persistence and produces a stable, low level of inflammation throughout the body, based on a small increase in immune system markers in the blood or tissues. At the human health level, one can simply analogize "normal-low grade inflammation-acute inflammation" to "healthy-sub-healthy-disease".
The existing research mainly aims at the regulation of low-grade inflammation and intestinal microbial disorder of targeted intestinal microorganisms, and intervenes the occurrence and development of low-grade inflammation and intestinal microbial disorder related to obesity by introducing prebiotics or probiotics, but the research is still in the development stage, and no product capable of realizing mass production is available.
Therefore, in order to solve the problem of the lack of products which can intervene in low-grade inflammation and intestinal microbial disorder related to obesity, an effective preparation with development prospect is urgently needed to meet the requirements of scientific research and markets.
Disclosure of Invention
The invention aims to provide the application of a nano-silver preparation in preparing a preparation for improving obesity-related low-grade inflammation caused by high-fat diet;
the invention also aims to provide the application of the nano-silver preparation in preparing a preparation for improving the intestinal flora disorder;
it is another object of the present invention to provide a regulator of obesity-related low grade inflammation for veterinary use;
it is another object of the present invention to provide a veterinary medicament for treating intestinal flora disorders.
The technical scheme adopted by the invention is as follows:
in a first aspect of the present invention, there is provided:
the application of the nano-silver preparation in preparing a preparation for improving obesity-related low-grade inflammation caused by high-fat diet; wherein, the components of the nano-silver preparation comprise PVP coated silver nano-particles (Ag NPs) with the concentration of 0.5-50 mg/L.
The nano silver has wide application in the field of biomedicine, has antibacterial property, can be applied to wound dressing and surface disinfection of medical instruments, and has unclear evaluation on the efficacy and biological safety of related products.
The nano silver used in the examples of the present invention is PVP coated silver nanoparticles with a particle size of 23.7 ± 5.5nm, and is available from guangzhou hongwu materials science and technology ltd.
High fat diets result in obesity, accompanied by low grade inflammation and intestinal microbial disturbances. Acute inflammation can be recovered quickly, while chronic low-grade inflammation usually has no obvious symptoms (and is therefore often ignored), and can only be accurately judged through the detection of a marker immune index at a molecular level, wherein C-reactive protein (CRP) and interleukin-6 (IL-6) are used as molecular indexes for characterizing low-grade inflammation in the invention. Moreover, researches also find that chronic low-grade inflammation plays a crucial role in the occurrence and development processes of various inflammatory diseases such as diabetes, heart disease, cancer, rheumatoid arthritis, senile dementia and the like, so that improvement of obesity-related low-grade inflammation is very important.
Further, the obesity-related low-grade inflammation is weight gain caused by high fat diet, increased serum triglyceride level, increased total cholesterol level, decreased glutamic-oxaloacetic transaminase/glutamic-pyruvic transaminase ratio, increased leukocyte count, and abnormal expression of C-reactive protein and interleukin-6.
Furthermore, the above C-reactive protein and interleukin-6 expression abnormalities include up-regulation of serum C-reactive protein and up-regulation of interleukin-6 expression.
In a second aspect of the present invention, there is provided:
application of nano-silver preparation in preparing preparation for improving intestinal flora disorder; the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
Further, the intestinal flora improvement disorder is improvement of intestinal microorganism abundance and inhibition of harmful bacteria activity.
Further, the improvement of the intestinal microbial abundance includes up-regulation of microbial abundance of phylum Vibrionaceae and deironium, and down-regulation of microbial abundance of phylum Epsilonbacteraeota and Actinomycetes.
Further, the above-mentioned activity against harmful bacteria includes an activity against Escherichia coli.
In a third aspect of the present invention, there is provided:
a regulator of obesity-related low-grade inflammation for animals is prepared from nano-silver; the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
In a fourth aspect of the present invention, there is provided:
a veterinary medicament for treating intestinal flora disorder is prepared from a nano-silver preparation; the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
The invention has the beneficial effects that:
1. the invention realizes the alleviation and the repair of low-degree inflammation and intestinal flora disorder induced by high-fat diet by the nano-silver preparation for the first time.
2. The nano-silver preparation has a minimum concentration which has a significant antibacterial effect (p <0.05) on typical gram-negative bacteria escherichia coli (E.coli) and is less than or equal to 5.0mg/L, does not show a significant growth inhibition effect on typical gram-positive bacteria staphylococcus aureus (S.aureus) under the condition of the highest test dose (50mg/L), and has a regulating effect on intestinal microorganisms of Verrucomicrobia (Verrucomicrobia), Deferribacter (Deferribacteria), Epsilon bacteria aeota and Actinobacillus (Actinobacillus).
3. The nano-silver preparation exposure of the invention significantly eases the mouse systemic low-grade inflammation response induced by high-fat diet and does not generate significant toxic effect on normal and obese mice.
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Coli, wherein vehicle represents a negative control without nanosilver;
fig. 2 is an in vitro antibacterial activity of nano-silver against typical gram-positive bacteria s.aureus, wherein vehicle represents a negative control without nano-silver;
FIG. 3 is a graph of the effect of long-term low-dose nanosilver exposure on body weight in normal and obese mice;
FIG. 4 is a graph of the effect of long-term low dose nanosilver exposure on serum Triglyceride (TG) (a), Total Cholesterol (TC) (b) and aspartate aminotransferase/glutamate pyruvate transaminase (AST/ALT) (c) levels in normal and obese mice, where ND represents a normal diet and HFD represents a high fat diet;
FIG. 5 is a graph of the effect of nanosilver exposure on the number of White Blood Cells (WBC) (a), C-reactive protein (CRP) (b) and interleukin-6 (IL-6) (C) expression in serum reflecting low levels of inflammation;
FIG. 6 is a graph of the effect of low dose nanosilver (5mg/L) exposure on mouse intestinal microbial abundance at the phylum classification level.
Detailed Description
In order to make the objects, technical solutions and technical effects of the present invention more clear, the present invention will be described in further detail with reference to specific embodiments. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.
The experimental materials and reagents used are, unless otherwise specified, all consumables and reagents which are conventionally available from commercial sources.
The nanosilver used in the examples described below was PVP coated silver nanoparticles 23.7 ± 5.5nm in size, available from guangzhou macrowu materials science and technology ltd.
Construction of obese mouse model
Male C57BL/6J mice (purchased from the laboratory animal center of Beijing military medical academy of sciences) of 5 weeks were adaptively raised for 1 week, and then fed with mouse growth and breeding feed (control group, mouse growth and breeding feed components: corn, wheat middling, soybean meal, flour, vegetable oil, calcium hydrogen phosphate, fish meal, choline chloride, additives, salt, antioxidant, purchased from Tay (Beijing) Biotech Co., Ltd.) and mouse 45% fat-energized high fat feed (test group, mouse 45% fat-energized high fat feed components: growth and breeding feed, lard, cholesterol, bile salt, sucrose, casein, premix and maltodextrin, purchased from Tay (Beijing) Biotech Co., Ltd.) respectively, high fat diet obese mouse models in the test group were prepared by continuously drinking water to which low-dose nano-silver (0, 0.5, 5, 50mg/L) was added, four groups of test groups of HFD0, HFD 0.5, HFD 5 and HFD 50 are respectively constructed; control group mouse model through continuously drinking water added with low dose of nano silver (0, 0.5, 5, 50mg/L), four groups of ND 0, ND 0.5, ND 5 and ND 50 control groups are respectively constructed to evaluate the regulating effect of nano silver on obesity-related low-grade inflammation and intestinal flora disorder. Mice were weighed and recorded every 2 weeks and sacrificed 24 weeks later.
1. Detection of in vitro antibacterial activity of nano-silver
E.coli (strain number: BL21(DE3) from Annu (Beijing) Biotechnology Co., Ltd.) and S.aureus (strain number: GDMCC1.1220 from Guangdong province culture Collection) were selected as representatives of gram-negative and gram-positive bacteria, respectively, and used for evaluating the antibacterial activity of the nano-silver against different bacteria.
The detection test of the in-vitro antibacterial activity of the nano-silver in the implementation comprises the following steps:
inoculating E.coli and S.aureus in logarithmic growth phase into LB culture medium containing different concentrations of nano-silver (0, 0.5, 5.0, 50mg/L), and diluting to specific concentration (OD)600nm0.06), culturing in a constant temperature incubator at 37 deg.C, sampling every 0.5 hr, and measuring OD of the sample600nmAnd (3) drawing a bacterial growth curve (shown in figure 1 and figure 2).
The results show that the lowest concentration of the nano-silver which has a significant bacteriostatic effect (p <0.05) on the typical gram-negative bacteria E.coli is less than or equal to 5.0mg/L (figure 1), and the nano-silver does not have a significant growth inhibition effect on the typical gram-positive bacteria S.aureus at the highest tested dose (50mg/L) (figure 2).
The results show that the nano-silver has different bacteriostatic activities on different types of microorganisms and is a potential intestinal microorganism regulator.
2. Toxicity test of nano-silver exposure to normal and obese mice
The test procedure was as follows:
the normal and fat mouse models constructed by the method are taken, mouse serum is separated, and the biochemical indexes of the mouse serum of each exposure group (test group and control group) are detected by an automatic serum biochemical analyzer (model: Chemray 240).
As a result:
the influence of the obtained high-fat diet feeding and nano-silver exposure on the body weight of normal and obese mice is monitored as shown in fig. 3, and the biochemical result of blood detection at the end point is shown in fig. 4, so that the results show that the body weight of the mice is remarkably increased, the blood lipid level (TG and TC) is increased, the AST/ALT ratio is reduced, and the nano-silver exposure of 0.5-50mg/L does not remarkably influence the biochemical indexes of the body weight and the blood serum for 24 weeks, which indicates that the nano-silver exposure does not have remarkable toxic effect on the normal and obese mice.
3. Nano-silver relieves systemic low-grade inflammation induced by high-fat diet
The test procedure was as follows:
collecting blood of normal and obese mice model constructed by the method, collecting blood in an anticoagulation tube containing EDTA salt, and detecting the amount of leucocyte by using a full-automatic blood cell analyzer (model: BC-2800 vet).
The normal and obese mouse models constructed by the method were used to isolate mouse serum, and CRP and IL-6 levels in the serum were measured using CRP and IL-6ELISA detection kits (purchased from Shanghai Jianglai Biotech, Inc. and Saimer Feishell science, Inc. (China), respectively).
By detecting the number of leucocytes in blood and the expression levels of C-reactive protein (CRP) and inflammatory factor IL-6 with low inflammation level in serum, the number of leucocytes in the blood of mice in an obese group (HFD0) can be obviously increased, and the levels of CRP and IL-6 in the serum are obviously higher than those of normal mice, which indicates that the obese mice are in a low-grade inflammation state. Low dose nanosilver exposure significantly alleviated the high fat diet-induced systemic low-grade inflammatory response described above (figure 5).
4. Nano-silver for repairing high-fat diet-induced intestinal microbial disorder
Mouse feces were collected 1 day before the mice constructed in the above-described procedure for constructing an obese mouse model, RNA was extracted, and microorganisms in the mouse feces of each exposure group (test group and control group) were analyzed by 16SRNA sequencing.
The results show that nanosilver exposure has a significant repairing effect on high fat diet-induced intestinal microbial disorders, in particular, high fat diet feeding significantly down-regulates the abundance of microorganisms of Verrucomicrobia (Verrucomicrobia, a in fig. 6), desferriobacteria (d in fig. 6), and up-regulates the abundance of microorganisms of epsilon bacillaeota (b in fig. 6) and actinomycetia (Actinobacteria, c in fig. 6), resulting in intestinal microbial disorders. And the low-dose nano-silver exposure has a remarkable repairing effect on the intestinal microbial disorder induced by high-fat diet (the abundance of microorganisms of the phylum of verrucomicrobia and deironium is adjusted up to be close to a normal group, and the abundance of microorganisms of the phylum of Epsilonbacteraeota and actinomycete is adjusted down to be close to a normal group).
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. The application of the nano-silver preparation in preparing a preparation for improving obesity-related low-grade inflammation caused by high-fat diet; wherein the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
2. The use according to claim 1, wherein the manifestations of high fat diet-induced obesity include weight gain, elevated triglyceride levels, elevated total cholesterol levels; the obesity-related low-grade inflammation is expressed by any one of an increase in the number of leukocytes, an abnormality in the expression of C-reactive protein, and an abnormality in the expression of interleukin-6 caused by obesity.
3. The use of claim 2, wherein said C-reactive protein expression abnormality comprises an upregulation of serum C-reactive protein expression and said interleukin-6 expression abnormality comprises an upregulation of serum interleukin-6 expression.
4. Application of nano-silver preparation in preparing preparation for improving intestinal flora disorder; wherein the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
5. The use of claim 4, wherein the intestinal flora improvement disorder is improvement of intestinal microbial abundance and inhibition of harmful bacterial activity.
6. The use of claim 5, wherein said improving gut microbial abundance comprises up-regulating microbial abundance of phyla wartomycetes and deinocentomomycota and down-regulating microbial abundance of phyla epsilon bacteraeota and actinomyceta.
7. The use of claim 5, wherein said inhibiting harmful bacterial activity comprises inhibiting escherichia coli activity.
8. A regulator of obesity-related low-grade inflammation for animals is characterized in that the regulator of obesity-related low-grade inflammation for animals is prepared from a nano-silver preparation; the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
9. A veterinary medicament for treating intestinal flora disorder is characterized by being prepared from a nano-silver preparation; the components of the nano-silver preparation comprise PVP coated silver nano-particles with the concentration of 0.5-50 mg/L.
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| CN104258394A (en) * | 2014-09-28 | 2015-01-07 | 中国科学技术大学 | Nano-silver combined autophagy inhibitor capable of killing and damaging tumor |
| US20200078397A1 (en) * | 2016-11-30 | 2020-03-12 | Xeno Biosciences Inc. | Pharmaceutical preparations and methods to manage weight and to modulate the gut microbiota |
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| CN104258394A (en) * | 2014-09-28 | 2015-01-07 | 中国科学技术大学 | Nano-silver combined autophagy inhibitor capable of killing and damaging tumor |
| US20200078397A1 (en) * | 2016-11-30 | 2020-03-12 | Xeno Biosciences Inc. | Pharmaceutical preparations and methods to manage weight and to modulate the gut microbiota |
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