CN115644451A - Mineral composition and product and application thereof - Google Patents
Mineral composition and product and application thereof Download PDFInfo
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- CN115644451A CN115644451A CN202211230454.9A CN202211230454A CN115644451A CN 115644451 A CN115644451 A CN 115644451A CN 202211230454 A CN202211230454 A CN 202211230454A CN 115644451 A CN115644451 A CN 115644451A
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Abstract
The present invention provides a mineral composition comprising: calcium ions, magnesium ions and sulfate ions. The composition has effects in improving intestinal barrier and permeability, and regulating expression of intercellular tight junction protein.
Description
Technical Field
The invention relates to the technical field of food, in particular to a mineral composition, a product and application thereof.
Background
Besides digestion and absorption, normal intestinal functions also have a strong barrier function for resisting harmful organisms in the intestinal tract and various toxins produced by the harmful organisms, and the intestinal health is ensured to mainly depend on three barriers of the intestinal tract, namely an intestinal mucosal epithelial barrier, an immune barrier formed by intestinal immune cells and secretions thereof and a biological barrier formed by normal microbial flora in the intestinal tract.
The existing scientists have made many researches on finding the regulating and controlling substances of the intestinal barrier, starting from functional saccharides, such as chitosan, which can improve the growth performance and the humoral immunity of early weaned animals by up-regulating IGF-artificial induced growth axis, relieving matrix oxidative stress and improving the matrix of the intestinal barrier. Starting from probiotics, intestinal epithelial cell metabolic activity and membrane integrity are protected, for example by oral administration of enterococcus faecium EF 1.
Therefore, it is highly desirable to provide a mineral composition that improves the intestinal barrier.
Disclosure of Invention
In view of the above, the present invention provides a mineral composition for improving intestinal barrier.
The present invention provides a mineral composition comprising: calcium ions, magnesium ions and sulfate ions.
Preferably, the mass ratio of the calcium ions to the magnesium ions to the sulfate ions is (1-25): 1: (0.01-16).
Preferably, the mass ratio of the calcium ions, the magnesium ions and the sulfate ions is (1.5-22): 1: (0.03-10).
Preferably, the source of calcium ions comprises one or more of calcium acetate, calcium caseinate, calcium chloride, calcium citrate malate, calcium gluconate, calcium lactate, calcium malate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium sulfate, calcium ascorbate, calcium glycerophosphate, or milk calcium;
the source of the magnesium ions comprises one or more of magnesium sulfate, magnesium chloride, magnesium carbonate, magnesium bicarbonate or magnesium gluconate;
sources of the sulfate ions include: one or more of zinc sulfate, ferrous sulfate, copper sulfate or manganese sulfate.
The invention provides application of the composition in any one of the technical schemes in preparing a product for improving intestinal barrier.
Preferably, the improving intestinal barrier comprises improving permeability, promoting expression of claudin and/or enhancing cell viability.
The invention provides a product for improving intestinal barrier, which comprises the composition of any one of the technical schemes.
Preferably, the concentration of calcium ions in the product is 0.9-14.4 mmol/L;
the concentration of magnesium ions in the product is 0.1-1.6 mmol/L.
Preferably, the concentration of sulfate ions in the product is 0.1-1.6 mmol/L.
Preferably, the product is a dairy product; the dairy products comprise infant formula milk powder, children formula milk powder and adult formula milk powder.
In contrast to the prior art, the present invention provides a mineral composition comprising: calcium ions, magnesium ions and sulfate ions. The composition has effects in improving intestinal barrier and permeability, and regulating expression of intercellular tight junction protein.
Drawings
FIG. 1 is a Caco-2 cell morphology map (X200) at different times in example 2 of the present invention;
FIG. 2 shows TEER values of the normal group and the model group as a function of time in example 3 (A) of the present invention; (B) normal and model group Papp cases;
FIG. 3 shows the Papp values of different processing groups in embodiment 4 of the present invention;
FIG. 4 is an immunofluorescence plot (x 200) for each panel of example 5 of the present invention.
Detailed Description
The invention provides a mineral composition and a product and application thereof, and a person skilled in the art can use the content to realize the mineral composition by appropriately improving process parameters. It is expressly intended that all such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the scope of the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The present invention provides a mineral composition comprising: calcium ions, magnesium ions and sulfate ions.
According to the invention, the mass ratio of the calcium ions, the magnesium ions and the sulfate ions is (1-25): 1: (0.01-16).
In a preferred embodiment of the present invention, the mass ratio of the calcium ion, the magnesium ion and the sulfate ion is (1.5 to 22): 1: (0.03-10).
In a preferred embodiment of the present invention, the mass ratio of the calcium ion, the magnesium ion and the sulfate ion is (1.5 to 21.5): 1: (0.04-8).
In a preferred embodiment of the present invention, the mass ratio of the calcium ion, the magnesium ion and the sulfate ion is (1.7 to 21.5): 1: (0.04-6).
In some preferred embodiments of the present invention, the mass ratio of calcium ions, magnesium ions, and sulfate ions is (1.88 to 21.5): 1: (0.04-5).
In the present invention, the source of calcium ions comprises one or more of calcium acetate, calcium caseinate, calcium chloride, calcium citrate malate, calcium gluconate, calcium lactate, calcium malate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium sulfate, calcium ascorbate, calcium glycerophosphate, or milk calcium;
the source of the magnesium ions comprises one or more of magnesium sulfate, magnesium chloride, magnesium carbonate, magnesium bicarbonate or magnesium gluconate;
sources of the sulfate ions include: one or more of zinc sulfate, ferrous sulfate, copper sulfate or manganese sulfate.
The inventor finds that the mineral composition provided by the invention can be used for improving intestinal barrier, and comprises the following components: repair damaged intestinal barriers, improve intestinal permeability (TEER and Papp values), regulate intercellular tight protein junction expression, and the like.
The components and the proportion of the invention are the technical characteristics that the functions mutually support and have interaction relation; the technical effects of the present invention can be achieved as long as the above-specified components and proportions are satisfied.
The invention provides the use of a composition according to any of the preceding claims for the preparation of a product for improving the intestinal barrier.
In particular, the improvement of intestinal barrier comprises improving permeability, promoting expression of claudin and/or enhancing cell viability.
The invention provides a product for improving intestinal barrier, which comprises the composition of any one of the technical schemes.
The products of the present invention include, but are not limited to, food and/or health care products.
The product of the invention is a dairy product;
the dairy product comprises liquid milk (pasteurized milk, sterilized milk, concocted milk and fermented milk); milk powder (whole milk powder, skimmed milk powder, partially skimmed milk powder, modified milk powder, and bovine colostrum powder); other dairy products. The method specifically comprises the following steps: the first is liquid milk. Mainly comprises sterilized milk, yoghourt and the like. The second type is the milk powder type. Including whole milk powder, skimmed milk powder, whole sweetened milk powder, flavored milk powder, infant milk powder and other formula milk powder. The third category is condensed milk. The fourth type is milk fat. Including cream for cake making, cream for common bread, and the like. The fifth category is cheeses and processed cheeses. The sixth category is the milk ice cream category. The seventh category is other dairy categories. Mainly comprises casein, lactose, milk tablets and the like.
The dairy product preferably comprises one or more of concocted milk, concocted milk powder, fermented milk, milk slices, cheese or milk snacks. More preferably, the dairy product comprises infant formula, children formula and adults formula.
The components and proportions of the composition are clearly described above, and are not described in detail herein.
The invention does not limit the rest ingredients or auxiliary materials of the dairy product, and the auxiliary materials are well known to those skilled in the art.
According to the invention, the concentration of calcium ions in the product is preferably 0.9-14.4 mmol/L; more preferably 0.9 to 12mmol/L; most preferably 0.9 to 10mmol/L; particularly preferably 0.9 to 3.6mmol/L;
the concentration of magnesium ions in the product is preferably 0.1-1.6 mmol/L.
The concentration of sulfate ions in the product is preferably 0.1-1.6 mmol/L.
In some preferred embodiments of the invention, the product comprises:
the concentration of calcium ions is 0.9-14.4 mmol/L, the concentration of magnesium ions is 0.1-1.6 mmol/L, and the concentration of sulfate ions is 0.1-1.6 mmol/L;
in some of the preferred embodiments of the invention, the product comprises:
the concentration of calcium ions is 0.9-3.6 mmol/L, the concentration of magnesium ions is 0.1-1.6 mmol/L, and the concentration of sulfate ions is 0.1-1.6 mmol/L;
in one of the preferred embodiments of the invention, the product comprises:
the concentration of calcium ions is 1.8mmol/L, the concentration of magnesium ions is 0.4mmol/L, and the concentration of sulfate ions is 0.4mmol/L;
in one of the preferred embodiments of the invention, the product comprises:
the concentration of calcium ions is 1.8mmol/L, the concentration of magnesium ions is 1.6mmol/L, and the concentration of sulfate ions is 1.6mmol/L;
the invention also provides a preparation method of the product for improving the intestinal barrier, which comprises the step of mixing the raw materials.
The intestinal barrier damage model is preferably constructed by culturing Caco-2 cells in a calcium-free culture medium.
The composition with the specific proportion and the specific concentration can repair the damaged intestinal tract and improve the barrier function of the intestinal tract.
The composition can ensure that the tightly-connected protein is in a honeycomb network structure, the edge of the tightly-connected protein is clear and can emit strong green fluorescence, the tightly-connected protein tightly wraps cell nucleuses and is in a paving stone shape, the cell gaps are small, and the compactness is high; namely, the expression of the intercellular tight junction protein can be regulated; and simultaneously improves the cell activity.
To further illustrate the present invention, a mineral composition and its products and uses provided by the present invention are described in detail below with reference to the examples.
Example 1Effect of combination of calcium ions, magnesium ions and sulfate radicals on intestinal barrier injury model
And (3) adopting a calcium-free culture medium to culture Caco-2 cells so as to construct an intestinal barrier damage model. After the Caco-2 cells are recovered, the cells are subcultured to grow into compact monolayer cells. When the transmembrane Resistance value (TEER) of the monolayer cell exceeds 400 omega cm 2 And then, the intestinal barrier model is established successfully. TEER values in excess of 400. Omega. Cm in Caco-2 model 2 And then, continuously culturing the normal group by using a normal group replacement solution (a normal DMEM culture medium), culturing the calcium-free group by using the calcium-free culture medium until the intestinal barrier is damaged, and detecting related indexes of each group. Each set was provided with 3 parallel holes. Media information is as follows: DMEM high-glucose medium, DMEM calcium ion-free medium, calcium ion-free Phosphate Buffer (PBS), fetal Bovine Serum (FBS): gibco, inc. of USA; 0.25% tryptic digest (EDTA-containing): beijing Solaibao science and technology, inc.;
after model construction was complete, the morphology of the cells of the Caco-2 model was observed and the transmembrane resistance of a monolayer of cells was measured using a Millicell-ERS 2 resistance meter. The apparent permeability coefficient (Papp) values in the model were then determined using fluorescein (free acid) from sigma aldrich.
Firstly, a CC-8 kit is utilized to respectively detect the influence of tested substances (calcium ions, magnesium ions and sulfate ions) on the survival rate of Caco-2 cells. The inoculum concentration in 96-well plates was 1.0X 10 5 The cell suspension is obtained per mL and placed in an incubator for culture, and Ca with different concentrations is added after the cells grow compactly 2+ The ion culture is continued for 4 days, then CCK-8 solution is added, the incubation is carried out for 2h in the dark, and the OD is measured 450nm And calculating the cell survival rate.
After constructing Caco-2 intestinal barrier model, ca-free was used 2+ After a Caco-2 cell is cultured by a culture medium to establish an injury model,and (4) setting a model group (continuously culturing in a calcium-free culture medium), adding culture solution with the concentration corresponding to the tested substance into the other groups, and measuring the related indexes after culturing for 4 d.
Grouping cells: model group, calcium group (Ca) 2+ Concentration: 1.8 mmol/L), calcium-Low magnesium-Low sulfate group (Ca) 2+ Concentration: 1.8mmol/L, mg 2+ Concentration: 0.4mmol/L, sulfate ion concentration: 0.4 mmol/L) and Ca-Homomagnesium-Homomosulfate group (Ca) 2 + Concentration: 1.8mmol/L, mg 2+ Concentration 1.6mmol/L, sulfate ion concentration 1.6 mmol/L)
The transmembrane resistance TEER and Papp values of each group were then determined. And finally, measuring the forms of the intercellular tight junction proteins ZO-1, occludin and Claudin-1 and the expression conditions of immune factors by adopting an immunofluorescence method.
Example 2: morphological observation of Caco-2 cells
In the model of example 1, caco-2 cells were smooth and cobblestone, with vacuoles in the middle of a few cells, and a tightly connected cell population was observed at 7 d. At 14d, the cells proliferated faster, a few dead cells fell off and floated, and the medium gradually became turbid. At 21d, the cells appeared irregular polygons and gradually fused into a monolayer of cells, increasing compactness. Indicating that a monolayer intestinal barrier model has formed at this point. FIG. 1 shows morphology diagrams (X200) of Caco-2 cells at different times.
Example 3: method for constructing intestinal barrier injury model by adopting calcium-free culture medium
The normal group and model group construction method of example 1 as described above, changes in TEER and Papp values of the normal group and model group were tracked to determine that the intestinal barrier injury model was completed. The TEER values of the calcium-free and normal groups trended similar and rising over time during the first 2 days after barrier formation, when the injury model was initiated. On day 3, however, the TEER values of the calcium-free group suddenly dropped below 100. Omega. Cm 2 And continues to descend; while TEER in the normal group still rose steadily. At 4d, the Papp values of the normal group cells were less than 1.0X 10-6cm/s, while the Papp values of the calcium-free group were significantly higher than that of the normal group (P)<0.01 ); FIG. 2 (A) Normal group and dieType group TEER values varied with time; (B) Normal and model group Papp cases
Example 4 Effect of calcium, magnesium and sulfate combinations on a model of intestinal Barrier Damage
Example 1 the effect of normal, model, calcium-high magnesium-high sulfate, and calcium-low magnesium-low sulfate on Papp values was determined and it was found that the Papp values of the model group were the highest, indicating the strongest intestinal permeability and the worst compactness, whereas the papps of the remaining 3 groups were significantly reduced compared to the model group, and the Papp values of the calcium-high magnesium-high sulfate group and the calcium-low magnesium-low sulfate group were not statistically different from the normal group (P > 0.05). The experimental result shows that the two test proportions can repair the damaged intestinal tract and improve the barrier function of the intestinal tract.
Example 5 different groups of intercellular tight junction protein profiles
The expression of the tight junction protein between groups of different examples 1 was determined by means of fluorescence immunoassay. The nucleus shrinkage of the model group is reduced, the three kinds of tight junction proteins are also shrunk into small rings, are scattered and distributed and emit weak green fluorescence, part of the tight junction proteins are broken or lost, and obvious large gaps are formed among cells, so that the damage degree of the cell barrier is high, and the compactness is low. The cell nucleuses of other groups are large and mellow, the boundary is clear, and strong dark blue fluorescence is emitted; the three closely-connected proteins are in a honeycomb network structure, the edges of the three closely-connected proteins are clear and visible, strong green fluorescence is emitted, the three closely-connected proteins tightly wrap cell nucleuses and are in a paving stone shape, the cell gaps are small, and the compactness is high.
Example 6 Effect of calcium, magnesium and sulfate ions on cell viability
To determine whether calcium and magnesium ions have an effect on cell viability, the effect of different concentrations of calcium and magnesium ions on cell viability was also determined experimentally. With Ca 2+ The survival rate of Caco-2 cells is in a trend of increasing first and then decreasing when the concentration gradient is increased. Three lower concentrations of Ca, 0.9, 1.8 and 3.6mmol/L 2+ Under the environment, the cell survival rate is obviously higher than that of the other concentration groups(P < 0.05), but the survival rate of the cells at these 3 concentrations was not significantly different (P > 0.05). According to the result, 1.8mmol/L (cell viability 75.88%) Ca was added 2+ Concentration as and subsequent to Mg 2+ And the compatible concentration of sulfate ions. Followed by 1.8mmol/L Ca 2+ Concentration and different concentrations of Mg 2+ And sulfate ions, the cell survival rate of all dosage groups exceeds 100 percent, wherein Mg 2+ And sulfate ion concentration of 1.6mmol/L, the cell survival rate is lowest but still exceeds 100%. According to this result, the two concentrations of 0.4mmol/L and 1.6mmol/L were set as the concentrations of magnesium and sulfate in the calcium-low magnesium-low sulfate group and the calcium-high magnesium-high sulfate group, respectively.
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.
Claims (10)
1. A mineral composition, comprising: calcium ions, magnesium ions and sulfate ions.
2. The composition according to claim 1, wherein the mass ratio of the calcium ions, the magnesium ions and the sulfate ions is (1 to 25): 1: (0.01-16).
3. The composition according to claim 2, wherein the mass ratio of the calcium ions to the magnesium ions to the sulfate ions is (1.5 to 22): 1: (0.03-10).
4. The composition according to any one of claims 1 to 3,
the source of the calcium ions comprises one or more of calcium acetate, calcium caseinate, calcium chloride, calcium citrate malate, calcium gluconate, calcium lactate, calcium malate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium sulfate, calcium ascorbate, calcium glycerophosphate or milk calcium;
the source of the magnesium ions comprises one or more of magnesium sulfate, magnesium chloride, magnesium carbonate, magnesium bicarbonate or magnesium gluconate;
sources of the sulfate ions include: one or more of zinc sulfate, ferrous sulfate, copper sulfate or manganese sulfate.
5. Use of a composition according to any one of claims 1 to 4 for the manufacture of a product for improving intestinal barrier.
6. The use according to claim 5, wherein improving gut barrier comprises improving permeability, promoting claudin expression and/or enhancing cell viability.
7. A product for improving gut barrier comprising the composition of any one of claims 1 to 4.
8. The product according to claim 7, wherein the concentration of calcium ions in the product is 0.9 to 14.4mmol/L;
the concentration of magnesium ions in the product is 0.1-1.6 mmol/L.
9. The product of claim 7, wherein the concentration of sulfate ions in the product is from 0.1 to 1.6mmol/L.
10. The product of claim 7, wherein the product is a dairy product; the dairy products comprise infant formula milk powder, children formula milk powder and adult formula milk powder.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000008955A1 (en) * | 1998-08-12 | 2000-02-24 | Kjell Reidar Thoresen | Synthetic milk mineral |
| JP2016029065A (en) * | 2015-09-18 | 2016-03-03 | 糧三 齋藤 | Composition for improving intestinal environment and intestinal tract barrier |
| CN108697823A (en) * | 2016-02-13 | 2018-10-23 | 段维新 | Solid solution, can be by the bone implant product of bio-absorbable and preparation and method for implantation |
| CN110623243A (en) * | 2019-09-11 | 2019-12-31 | 内蒙古蒙牛乳业（集团）股份有限公司 | High calcium salt compound and preparation method thereof |
| CN110720628A (en) * | 2019-11-07 | 2020-01-24 | 上海奥医生物医药科技有限公司 | Special clinical nutrition formula for protecting chemotherapy-damaged intestinal mucosa barrier function and preparation method thereof |
| CN113613718A (en) * | 2019-03-26 | 2021-11-05 | 三得利控股株式会社 | Composition for improving intestinal barrier function |
| CN114947138A (en) * | 2022-05-25 | 2022-08-30 | 河南磐康健康管理股份有限公司 | Milk calcium composition for preventing and improving calcium deficiency disease, and preparation method and application thereof |
-
2022
- 2022-09-30 CN CN202211230454.9A patent/CN115644451A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000008955A1 (en) * | 1998-08-12 | 2000-02-24 | Kjell Reidar Thoresen | Synthetic milk mineral |
| JP2016029065A (en) * | 2015-09-18 | 2016-03-03 | 糧三 齋藤 | Composition for improving intestinal environment and intestinal tract barrier |
| CN108697823A (en) * | 2016-02-13 | 2018-10-23 | 段维新 | Solid solution, can be by the bone implant product of bio-absorbable and preparation and method for implantation |
| CN113613718A (en) * | 2019-03-26 | 2021-11-05 | 三得利控股株式会社 | Composition for improving intestinal barrier function |
| CN110623243A (en) * | 2019-09-11 | 2019-12-31 | 内蒙古蒙牛乳业（集团）股份有限公司 | High calcium salt compound and preparation method thereof |
| CN110720628A (en) * | 2019-11-07 | 2020-01-24 | 上海奥医生物医药科技有限公司 | Special clinical nutrition formula for protecting chemotherapy-damaged intestinal mucosa barrier function and preparation method thereof |
| CN114947138A (en) * | 2022-05-25 | 2022-08-30 | 河南磐康健康管理股份有限公司 | Milk calcium composition for preventing and improving calcium deficiency disease, and preparation method and application thereof |
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