阅读说明：本技术 一种含有水飞蓟提取物和后生元的组合物及其在护肝中的应用 (A composition containing herba Silybi Mariani extract and metazoan, and its application in protecting liver ) 是由 丁峰 李靖 于 2021-09-13 设计创作，主要内容包括：本发明公开了一种组合物,组合物含有水飞蓟提取物和后生元；水飞蓟提取物可以为水飞蓟提取物微胶囊形式,后生元可以选择为灭活的植物乳杆菌L137,后生元可以调节肠道菌群平衡,修复肠道,减少肠漏降低对肝炎的诱因,水飞蓟提取物与后生元的组合具有减少肝损伤的功效。(The invention discloses a composition, which contains silybum marianum extract and metazoan; the herba Silybi Mariani extract can be in the form of microcapsule, and the metazoan can be inactivated Lactobacillus plantarum L137, and can regulate intestinal flora balance, repair intestinal tract, reduce intestinal leakage, reduce cause of hepatitis, and reduce liver injury.)

1. A composition characterized by: contains herba Silybi Mariani extract and prebiotics.

2. The composition of claim 1, wherein: the herba Silybi Mariani extract comprises one or more of Silibinin A, Silibinin B, isosilybin A, isosilybin B, silychristin, isosilychristin, silydianin, and taxifolin;

and/or

One or more of silybin A, silybin B, isosilybin A, isosilybin B, silychristin, isosilychristin, silydianin, and taxifolin; thioalkyl lipid derivatives are preferred.

3. The composition of claim 1, wherein: the silybum marianum extract is in a silybum marianum extract microcapsule form and/or a silybum marianum extract liposome form and/or a silybum marianum extract self-microemulsifying form, and is preferably in one or more of a silybum marianum extract-phospholipid form, a silybum marianum extract-phospholipid-cholesterol form and a silybum marianum extract-cyclodextrin form; further preferably, the silybum marianum extract-phospholipid-cholesterol liposome form and the silybum marianum extract-phospholipid complex form are compounded, or the silybum marianum extract-phospholipid-cholesterol liposome form and the silybum marianum extract-cyclodextrin form are compounded.

4. The composition according to claim 1, characterized in that: the postbiotic is prepared from lactobacillus plantarum; further, lactobacillus plantarum L137 was used.

5. The composition according to claim 1, characterized in that: the content of herba Silybi Mariani extract in the composition is 1-99.9% (w/w), preferably 10-99.5% (w/w), and more preferably 90-99% (w/w); the content of Lactobacillus plantarum is 0.01-99% (w/w), preferably 0.05-90% (w/w), and more preferably 1-10% (w/w);

and/or, the composition may or may not contain a filler/carrier, and the amount of lactobacillus plantarum in the composition is 0.5-500 billion/g, further 1-100 billion/g, further 10-50 billion/g, further 20-30 billion/g.

6. The composition according to claim 1, characterized in that: the content of silibinin in the herba Silybi Mariani extract is more than 20%, preferably more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, more than 96%, more than 97%, more than 98%, more than 99% or 100%.

7. A product comprising the composition of any one of claims 1 to 6, wherein the product is in the form of one of injection, oral liquid, tablet, hard capsule, soft capsule, candy, solid beverage, liquid beverage, and soft extract.

8. The product of claim 7, further comprising an adjuvant; wherein the auxiliary materials comprise: water, excipient, pigment, sweetener, pH regulator, thickener, binder, disintegrating agent, and filler.

9. Use of the composition according to any one of claims 1 to 6, the product according to claims 7, 8 in a liver-protecting pharmaceutical product, a nutraceutical product, a dietary supplement and/or a food product;

or/and in anti-fatigue drugs, health foods, dietary supplements and/or foods;

or/and in antidepressant and anxiolytic drugs, health food, dietary supplements and/or food.

Technical Field

The invention relates to a composition, in particular to a composition with the effect of reducing liver injury and the like and application thereof.

Background

Silybum marianum, also known as milk thistle, originally produced in the Mediterranean region has been used as a medicinal plant for nearly two thousand years and is still widely used as a very effective liver-protecting plant. It can help the liver detoxify, and can also be used to control chronic liver disease. Silybum marianum can activate regeneration of liver protein, recover activity of liver cell destroyed by alcohol and virus, scavenge free radical, maintain permeability of liver cell membrane, reduce damage of toxin to liver, and protect people with liver injury due to long-term drinking and acute and chronic hepatitis, fatty liver, liver cirrhosis and toxic liver injury.

The silybum marianum extract is prepared from dried fruits of silybum marianum, and the commercial extract is generally standardized to contain 80% of total silymarin. Silymarin is a complex mixture of polyphenols, including seven closely related flavonol-lignin adducts (silybin a, silybin B, isosilybin a, isosilybin B, silychristin, isosilychristin, and silydianin) and a flavonoid taxifolin (taxifolin). Silibinin is the semi-purified fraction of silymarin, mainly a mixture of two diastereomers (silibinin a and silibinin B).

When we are in a healthy state, the intestinal wall of the small intestine only allows nutrients to pass through the intestinal wall into the blood, through the blood circulation to other parts of the body where it is needed. Other substances that are not allowed to enter are uniformly rejected. The intestinal wall cells are connected with each other by tight junctions, however, the tight junctions are easily damaged, such as obesity, chronic fatigue, dysbacteriosis, etc.

For sub-healthy people with high stress, obesity, metabolic disorders, intestinal disorders and the like, intestinal cells and tight junction proteins are damaged, and gaps between cells become larger, so that lipopolysaccharide derived from microorganisms or food passes through the intestinal wall and binds with lipoprotein to enter liver circulation, thereby causing liver inflammation. For sub-health people, the supplement of antibacterial, anti-inflammatory and antioxidant such as silybum marianum can only reduce the inflammation which is already generated, and the supplement cannot play a role in protecting the liver inflammation caused by the externally invaded lipopolysaccharide and the like.

The invention combines silybum marianum and metazoan, thereby playing roles of preventing and curing liver diseases.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provide a composition containing silybum marianum extract and prebiotics.

Further, the herba Silybi Mariani extract comprises one or more of Silibinin A, Silibinin B, isosilybin A, isosilybin B, silychristin, isosilychristin, silydianin, and taxifolin; it is further preferred that the silybum marianum extract comprises a combination of silybin a and other types of silybum marianum extracts.

Furthermore, the silybum marianum extract is in a silybum marianum extract microcapsule form and/or a silybum marianum extract liposome form and/or a silybum marianum extract self-microemulsifying form, and is preferably in one or more of a silybum marianum extract-phospholipid form, a silybum marianum extract-phospholipid-cholesterol form and a silybum marianum extract-cyclodextrin form.

Further preferably, the silybum marianum extract-phospholipid-cholesterol liposome form and the silybum marianum extract-phospholipid complex form are compounded, or the silybum marianum extract-phospholipid-cholesterol liposome form and the silybum marianum extract-cyclodextrin form are compounded.

Further, the metazoan is dead bacteria, or comprises live bacteria and dead bacteria. Preferably, the microorganism contains at least dead cells. In the combination of dead cells and live cells, the ratio of dead cells to live cells was 99:1, 95:5, 90:10, 80:20, 75:25, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90, 1: 99.

Furthermore, metazoan contains bacteria and metabolites of bacteria.

Further, the metazoan is a metazoan made of lactobacillus plantarum.

Further, the lactobacillus plantarum is lactobacillus plantarum L137.

Further, Lactobacillus plantarum is a dead cell of L-137.

Further, the content of the extract of Silybum marianum in the composition is 1-99.9% (w/w), preferably 10-99.5% (w/w), and more preferably 90-99% (w/w); the Lactobacillus plantarum is 0.01-99% (w/w), preferably 0.05-90% (w/w), and more preferably 1-10% (w/w).

Furthermore, the silybin (silybin A and silybin B) in the silybum marianum extract accounts for more than 20% of the total content of silybin A, silybin B, isosilybin A, isosilybin B, silychristin, isosilychristin, silydianin, and taxifolin, and has a content of more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, more than 96%, more than 97%, more than 98%, more than 99% or 100%.

The purity of herba Silybi Mariani extract is above 50%, and has a purity of above 60%, above 70%, above 80%, above 90%, above 95%, above 96%, above 97%, above 98%, above 99% or 100%.

Further, the dead bacteria can be inactivated by heating, ultraviolet irradiation and chemical treatment; preferably, the cells are heat-inactivated. The heat-inactivated cells may be those which are collected after completion of the culture and then thermally killed, or they may be collected after the cells in the culture are made into the thermally killed cells without temporarily separating the cells from the culture. The heating temperature of the heated dead cells of Lactobacillus plantarum L-137 is usually about 60 to 100 ℃, preferably about 70 to 90 ℃, and a known means using a heater can be used as the heating means. The heating time after reaching the desired temperature is generally about 5 to 40 minutes, preferably about 10 to 30 minutes.

The dead cells may be further subjected to grinding, crushing, freeze-drying or the like to prepare a treated product of dead cells. In the present invention, the treated dead bacteria can be suitably used as dead bacteria.

The form of the dead cells in the present invention may be a paste form, a suspension form, a milk form, a powder form, or a granule form, and is preferably a powder form.

The number of dead bacteria in the invention is the number of dead bacteria after the number of live bacteria is inactivated. In the present invention, the number of dead cells can be estimated and determined by detecting the number of viable cells and the viable cell density of the dead cells, because of the limitation of the means for detecting the number of dead cells.

Further, the composition may or may not contain a filler/carrier, and the amount of Lactobacillus plantarum in the composition is 0.5 to 500, further 1 to 100, further 10 to 50, further 20 to 30 parts per million/g.

Further, the composition also contains glutathione.

The invention also provides a product which comprises the composition, and the dosage form of the product is one of injection, oral liquid, tablets, hard capsules, soft capsules, candies, solid beverages, liquid beverages and soft extracts.

Further, the product also comprises auxiliary materials; wherein the auxiliary materials comprise: water, excipient, pigment, sweetener, pH regulator, thickener, binder, disintegrant and filler.

The invention also provides the application of the composition in liver injury medicines, health-care foods, dietary supplements and foods;

or/and application in liver-protecting medicines, health-care foods, dietary supplements and foods.

Interpretation of terms

"anti", as used herein, unless otherwise specified, includes prophylactic, inhibitory, and scavenging effects.

The term "liver injury" as used herein includes, unless otherwise specified, injuries caused by physical, chemical, biological, and the like.

The "dead cells" used herein are, unless otherwise specified, inactivated cells.

The term "metazoan" as used herein refers to a general term for probiotic components after processing of probiotics, and includes bacteria and metabolites.

"L137", as used herein, is, unless otherwise specified, inactivated Lactobacillus plantarum L137.

Description of the drawings:

FIG. 1 shows ALT test condition of liver-protecting effect in animal test 1;

fig. 2 shows the AST test in liver-protecting effect animal test 2.

Detailed Description

The present invention will be further described with reference to the following examples for facilitating understanding of those skilled in the art, and the description of the embodiments is not intended to limit the present invention.

Example 1

A composition comprises Silybum marianum extract and inactivated Lactobacillus plantarum L137, wherein silybin (including silybin A and silybin B) accounts for 80% of the Silybum marianum extract, the Silybum marianum extract is in the form of Silybum marianum-phospholipid complex, the Silybum marianum extract is 40% (w/w), and inactivated Lactobacillus plantarum L137 is 60% (w/w).

Example 2

A composition comprises Silybum marianum extract and inactivated Lactobacillus plantarum L137, wherein silybin in the Silybum marianum extract accounts for 70%, the Silybum marianum extract accounts for 20% (w/w), and the inactivated Lactobacillus plantarum L137 accounts for 80% (w/w).

Example 3

A composition comprises Silybum marianum extract, inactivated Lactobacillus plantarum L137 and live Lactobacillus plantarum, wherein silybin in the Silybum marianum extract accounts for 90%, Silybum marianum extract accounts for 33.3% (w/w), and Lactobacillus plantarum accounts for 66.7% (w/w).

Example 4

A composition comprises Silybum marianum extract and inactivated Lactobacillus plantarum L137, wherein silybin in the Silybum marianum extract accounts for 80% of the Silybum marianum extract, the Silybum marianum extract accounts for 40% (w/w), the inactivated Lactobacillus plantarum L137 accounts for 40% (w/w), and glutathione accounts for 20% (w/w).

Example 5

A composition comprises Silybum marianum extract and inactivated Lactobacillus plantarum L137, wherein silybin in the Silybum marianum extract accounts for 80%, the Silybum marianum extract accounts for 99% (w/w), and the inactivated Lactobacillus plantarum L137 accounts for 1% (w/w).

Example 6

A composition comprises Silybum marianum extract and inactivated Lactobacillus plantarum L137, wherein silybin in the Silybum marianum extract accounts for 80%, Silybum marianum extract accounts for 10% (w/w), and inactivated Lactobacillus plantarum L137 accounts for 90% (w/w).

Example 7

A capsule comprises herba Silybi Mariani extract and inactivated Lactobacillus plantarum L137, wherein silybin accounts for 80% of the herba Silybi Mariani extract, herba Silybi Mariani extract is 0.12g, and inactivated Lactobacillus plantarum L137 is 2.4 mg.

Example 8

A capsule comprises herba Silybi Mariani extract and inactivated Lactobacillus plantarum L137, wherein silybin accounts for 80% of the extract, the extract is in the form of silybum marianum extract-phospholipid-cholesterol liposome, the extract is 0.06g, and the inactivated Lactobacillus plantarum L137 is 1.2 mg.

Example 9

A capsule comprises herba Silybi Mariani extract and inactivated Lactobacillus plantarum L137, wherein silybin accounts for 80% of the herba Silybi Mariani extract, herba Silybi Mariani extract is 0.24g, and inactivated Lactobacillus plantarum L137 is 4.8 mg.

Example 10

A capsule comprises herba Silybi Mariani extract and inactivated Lactobacillus plantarum L137, wherein silybin accounts for 60% of the herba Silybi Mariani extract, herba Silybi Mariani extract is 0.08g, and inactivated Lactobacillus plantarum L137 is 0.8 mg.

Experimental example 1

Animals: male Kunming mice (20 ± 2 g): mice were placed in an animal laboratory and incubated at room temperature. And (5) breeding in cages.

Liver-protecting effect animal test 1:

grouping: the 50 mice were randomly and equally divided into 5 groups, which were: control group, lipopolysaccharide group (liver injury group), lipopolysaccharide + silybum marianum group, lipopolysaccharide + L137 group, and lipopolysaccharide + silybum marianum + L137 group.

Control group: the feed and normal saline are fed for 15 days, and the normal saline is injected into the abdominal cavity.

Lipopolysaccharide group (liver injury group): the feed and normal saline are fed for 15 days, and 4mg/kg of Escherichia coli lipopolysaccharide is injected into the abdominal cavity.

Lipopolysaccharide + silybum marianum group: the feed, 75mg/kg silymarin and physiological saline were fed for 15 days, and 4mg/kg Escherichia coli lipopolysaccharide was injected intraperitoneally.

Lipopolysaccharide + L137 group: the feed, 1.5mg/kg L137 and normal saline are fed for 15 days, and 4mg/kg Escherichia coli lipopolysaccharide is injected into the abdominal cavity.

Lipopolysaccharide + silybum marianum + L137 group: the feed, 75mg/kg, 1.5mg/kg L137 and normal saline are fed for 15 days, and 4mg/kg Escherichia coli lipopolysaccharide is injected into the abdominal cavity.

After eating for 12 hours, blood was taken from the mice, centrifuged at 3000X g at 4 ℃ for 15 minutes, and the supernatant was taken for testing. Alanine aminotransferase ALT and aspartate aminotransferase AST were tested using a 756P uv-vis spectrophotometer.

TABLE 1 liver-protective Effect ALT and AST test conditions in animal test 1

	ALT(IU/L)	AST(IU/L)
			Control group	48.4±5.1	75.5±8.6
Lipopolysaccharide group (model group)	102.3±10.7,	169.7±19.3，
			Lipopolysaccharide and silybum marianum	85.7±9.4,p<0.05	127.7±15.9,p<0.05
Lipopolysaccharide + L137	95.1±9.8,p<0.05	159.2±13.1,p<0.05
			Lipopolysaccharide + Silybum marianum + L137	69.6±7.4,p<0.05	108.8±12.3,p<0.05

The data show that compared with the lipopolysaccharide group, the L137 group serum ALT and AST are reduced, and the silybum marianum + L137 group serum ALT and AST under the protection effect are obviously reduced. The reduced value of the silybum marianum + L137 composition is better than either silybum marianum or L137 alone.

It is demonstrated that mice taking silybum marianum + L137 daily have a stronger resistance to liver inflammation caused by toxins such as lipopolysaccharide. The silybum marianum extract can optimize mitochondrial metabolism, reduce the active oxygen level in cells and reduce the expression of NF-kB; and L137 blocks TLR4 mRNA, thereby affecting a TLR 4-NF-kB pathway, inhibiting NF-kB expression in liver, enhancing NF-kB reduction and enhancing the anti-inflammatory effect of silybum marianum.

Experimental example 2

40 male New Zealand white rabbits (957) -992g, 10 of which were used as control groups;

control group: the feed and the normal saline are fed every day, and the normal saline is injected into the abdominal cavity for two weeks.

Lipopolysaccharide group (model group): daily feed plus saline was fed, and 5mcg/kg of Escherichia coli lipopolysaccharide was intraperitoneally injected for two weeks.

After molding, the mixture was centrifuged at 3000X g and 4 ℃ for 15 minutes to obtain a supernatant. Alanine aminotransferase ALT and aspartate aminotransferase AST were tested using a 756P uv-vis spectrophotometer.

Control group: the feed is continuously fed with the daily feed and the physiological saline for six weeks.

The model components were divided into three groups, each serving as baseline.

Lipopolysaccharide + group L-137: daily feed +0.5mg/kg L137+ physiological saline was fed, and 0.5mcg/kg lipopolysaccharide was injected into the abdominal cavity for six weeks.

Lipopolysaccharide + silybum marianum group: daily feed +25mg/kg silymarin + physiological saline was fed and 0.5mcg/kg lipopolysaccharide was injected intra-abdominally for six weeks.

Lipopolysaccharide + silybum marianum + L137 group: daily feed +25mg/kg silymarin +0.5mg/kg L137+ physiological saline is fed, and 0.5mcg/kg lipopolysaccharide is injected into the abdominal cavity for six weeks.

Blood samples were taken from rabbits weekly, centrifuged at 3000X g, 4 ℃ for 15 minutes, and the supernatant was taken for testing. Alanine aminotransferase ALT and aspartate aminotransferase AST were tested using a 756P uv-vis spectrophotometer.

TABLE 2 liver-protecting Effect ALT test Condition of animal test 2

Table 3 liver-protecting effect test conditions of AST of animal test 2

As can be seen from tables 2-3 and fig. 2, in the case of liver inflammation caused by lipopolysaccharide and in the presence of interference of exogenous lipopolysaccharide, the rabbit using silybum marianum + L137 has a higher slope of the curve, i.e., inflammation can be reduced more quickly and the rabbit can take effect more quickly. On one hand, the L137 influences a metabolic pathway to further influence the anti-inflammatory effect of the silybum marianum, and on the other hand, the regulation effect of the L137 on intestinal flora is favorable for repairing intestinal leakage and reducing the damage of exogenous lipopolysaccharide to liver caused by entering a circulatory system through the intestinal tract. Meanwhile, the silybum marianum has the functions of diminishing inflammation and resisting bacteria to the liver, and the composition has the synergistic protective effect inside and outside.

It will be understood by those skilled in the art that, unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the prior art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.

It should be understood that the above detailed description of the technical solution of the present invention with the help of preferred embodiments is illustrative and not restrictive. On the basis of reading the description of the invention, a person skilled in the art can modify the technical solutions described in the embodiments, or can substitute part of the technical features of the embodiments equivalently; and the modifications or the substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.