Composition for treating or preventing metabolic disease, containing, as active ingredient, extracellular vesicles derived from akkermansia muciniphila bacteria
vesicles, which is applied in the field of pharmaceutical and food compositions containing extracellular vesicles derived from akkermansia muciniphila bacteria, can solve the problems of not knowing the mechanism of inhibiting the local inflammatory response caused by pathogenic bacteria-derived vesicles, no research finding that shows a metabolic disease such as obesity or diabetes can be alleviated or treated, etc., to achieve akkermansia muciniphila muciniphila muciniphila a technology of akkermansia muciniphila a technology of akkermansia muciniphila a technology of akkermansia muciniphila a technology of a metabolic disease and a technology of a metabolic disease and a technology of a metabolic disease and a technology of a drug and food composition and a technology of akkermansia muciniphila bacteria drug
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example 1
Isolation of Extracellular Vesicles from Culture Medium of Akkermansia Muciniphila
[0056]An Akkermansia muciniphila strain (ATCC BAA-835) was cultured in 2 L of autoclaved Brain-heart infusion broth (BD 237500) in an anaerobic chamber for 72 hours so that an O.D value became 1.5, and then a culture medium was centrifuged at a high speed (10,000×g) for 20 minutes to obtain a supernatant excluding a precipitated bacterial cell pellet. The supernatant was filtered with a 0.45 μm filter and a 0.22 μm filter, thus obtaining a culture medium concentrated about 14 fold using the Quixstand benchtop system. The culture medium was ultracentrifuged (150,000 ×g) at 4° C. for 2 hours to obtain pellets, and then the pellets was dissolved with sterile saline (PBS), followed by protein quantification.
example 2
Characteristic Analysis of Extracellular Vesicles
[0057]In order to confirm whether the substance obtained by the method of Example 1 is Akkermansia muciniphila-derived extracellular vesicles, a 50 μg / ml sample obtained by quantifying proteins was observed through the JEM 1011 electron microscope (commercially available from Jeol Ltd., Japan). As a result, as shown in FIG. 1, it can be seen that the Akkermansia muciniphila-derived extracellular vesicle is spherical.
[0058]In addition, in order to confirm a size of an extracellular vesicle isolated from Akkermansia muciniphila using a dynamic light scattering (DLS) method, a 50 μg / ml sample was measured through the Zetasizer Nano ZS (commercially available from Malvern Instruments Ltd, UK). As a result, as shown in FIG. 2, it can be seen that Akkermansia muciniphila-derived extracellular vesicles have an average diameter of 20 to 300 nm.
[0059]In addition, in order to confirm a protein expression pattern of extracellular vesicles isolat...
example 3
Therapeutic Effects of Akkermansia Muciniphila-Derived Extracellular vesicles in obese and diabetic mouse induced by a high fat diet
[0061]In order to confirm whether the Akkermansia muciniphila-derived extracellular vesicles obtained in Example 1 inhibit obesity and diabetes induced by a high fat diet, an experiment was performed as follows.
[0062]The extracellular vesicles obtained in Example 1 were administered to each of normal mice which were fed a regular diet (RD) for 2 months and obese and diabetic mouse models induced by being fed a high fat diet (HFD) for 2 months at a dose of 10 μg / mouse for 3 weeks at two day intervals (See FIG. 5). In this case, a group of only RD-fed mice and a group of RD-fed and extracellular vesicle-administered mice were used as negative controls.
[0063]In a result obtained by measuring weight while extracellular vesicles are administered at two day intervals, as shown in FIG. 6, it can be seen that a weight was not increased for 3 weeks in a HFD-fed,...
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