An immune function enhancing composition

Abstract

The application discloses an immune function enhancing composition which is mixed by sambucus williamsii alcohols extract and lactobacillus rhamnosus agent. The application can effectively immunize the spleen, improve the spleen index and thymus index, has the function of immune regulation, and can also improve the IgM and IgG content in serum.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of natural product application, and particularly relates to an immune function enhancing composition. BACKGROUND

[0002] Sambucus nigra L. is the most common species belonging to Adoxaceae family and Sambucus genus, and is widely distributed in Europe, West Asia, North America and the United States. The flowers, fruits and leaves of Sambucus nigra L. contain rich nutritional ingredients and bioactive substances such as carbohydrates, proteins, fatty acids, organic acids, minerals, vitamins, anthocyanins, polyphenols and flavonoids, and have the effects of antioxidant, antibacterial, anti-inflammatory and the like. It is traditionally a herb with different effects. The fruits of Sambucus nigra L. are often called elderberry.

[0003] As an important probiotic in the intestinal tract, probiotic lactic acid bacteria can induce the proliferation of immune cells and the secretion of immunologically active substances such as immunoglobulin A (IgA) and immunoglobulin G (IgG) through its own active substances such as teichoic acid. IgA and IgG are also important immunoglobulins in the body, which play an important role in resisting bacterial and viral infections, preventing the combination of symbiotic bacteria and epithelial cells, and balancing the process of toxins on the mucosal surface, and can effectively prevent the occurrence of corresponding infectious diseases. At the same time, probiotic lactic acid bacteria can also improve the immune-related metabolites in the intestinal tract, and induce immune cells to produce different immune responses when exposed to different infections, so as to regulate the immune capacity of the body. The main intestinal metabolites such as short chain fatty acids (SCFAs), amino acids and organic acids can interact with the immune system of the body, thereby regulating the immune capacity of the body.

[0004] Immunity is the ability of the body to recognize and eliminate foreign invaders such as viruses and bacteria, and to deal with its own aging, damaged, dead and degenerative cells. The intestine, as the largest immune organ of humans, contains most of the lymphocytes, macrophages and immune factors; lymphocytes can proliferate or produce bioactive substances under the stimulation of many non-specific substances; macrophages not only can initiate specific immune response, but also can effectively phagocytize harmful bacteria; lymphocytes and macrophages can also secrete various cytokines such as interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α), further promoting the differentiation and maturation of immune cells and improving the immune capacity of the body. With the continuous colonization of microorganisms in the intestine, the immune system begins to develop and gradually matures, and the colonized intestinal microorganisms not only help the body to digest, absorb and metabolize nutrients, but also can regulate the immune capacity of the body by generating signals that promote immune response. Research has found that the decline of the body's immunity is not only due to the changes of the body's genes, the lack of nutrition and exercise, but also the metabolic disorder of intestinal microorganisms is an important factor leading to low immunity of the body. SUMMARY

[0005] The present application aims to provide an immune function enhancing composition.

[0006] Another object of the present application is to provide the use of a combination of elderberry ethanol extract and Lactobacillus rhamnosus agent in the preparation of an immune function enhancing composition.

[0007] The technical solution of the present application is as follows:

[0008] An immune function enhancing composition is prepared by mixing elderberry ethanol extract and Lactobacillus rhamnosus agent, wherein,

[0009] The preparation of the elderberry ethanol extract comprises: drying and oil-removing the elderberry, crushing it into powder, then refluxing with 95% ethanol, filtering and drying by concentration to obtain the elderberry ethanol extract.

[0010] The Lactobacillus rhamnosus agent is the freeze-dried bacterial body of Lactobacillus rhamnosus.

[0011] In a preferred embodiment of the present application, the mass ratio of the elderberry ethanol extract and the Lactobacillus rhamnosus agent is 1-5:0.5-5.

[0012] Further preferably, the mass ratio of the elderberry ethanol extract and the Lactobacillus rhamnosus agent is 1:1.

[0013] In a preferred embodiment of the present application, the Lactobacillus rhamnosus is Lactobacillus rhamnosus CICC 6161.

[0014] The use of the combination of the Sambucus williamsii Hance ethanol extract and the Lactobacillus rhamnosus agent in the preparation of an immune function enhancing composition.

[0015] In a preferred embodiment of the present application, the preparation of the Sambucus williamsii Hance ethanol extract comprises: drying and oil-removing the Sambucus williamsii Hance, crushing into powder, refluxing with 95% ethanol, filtering and drying by concentration to obtain the ethanol extract.

[0016] Further preferably, the Lactobacillus rhamnosus agent is a freeze-dried bacterial body of Lactobacillus rhamnosus.

[0017] More preferably, the Lactobacillus rhamnosus is Lactobacillus rhamnosus CICC 6161.

[0018] In a preferred embodiment of the present application, the mass ratio of the Sambucus williamsii Hance ethanol extract and the Lactobacillus rhamnosus agent is 1-5:0.5-5.

[0019] Further preferably, the mass ratio of the Sambucus williamsii Hance ethanol extract and the Lactobacillus rhamnosus agent is 1:1.

[0020] The present application has the following advantages: the present application can effectively immunize the spleen, increase the spleen index and the thymus index, has the function of immune regulation, and can also increase the IgM and IgG contents in serum. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Figure for the weight change test results of the mice in Example 1 of the present application. DETAILED DESCRIPTION

[0022] The technical solutions of the present application are further described and explained in the following detailed description with reference to the accompanying drawings.

[0023] Example 1

[0024] (1) Preparation of the Sambucus williamsii Hance ethanol extract: Sambucus williamsii Hance was selected as the raw material, dried in a 65℃ oven, and most of the oil was removed by an oil press. The Sambucus williamsii Hance residue after oil pressing was crushed into powder by a powdering machine, and the powder was passed through a 60-mesh sieve and placed in a sealed bag for use. The powder was refluxed with 95% ethanol for 3 times, 1.5h each time, filtered, and the extract was combined and dried by concentration to obtain the ethanol extract, which was used as the test drug in the present study.

[0025] (2) Preparation of Lactobacillus rhamnosus agent: Lactobacillus rhamnosus CICC6161 (purchased from China Industrial Microbial Culture Collection Center) was activated and inoculated in MRS liquid medium, and then cultured at 37°C for 24h. The bacterial body was obtained by centrifugation at 12000r / min for 10min at 4°C. The bacterial body was washed twice with PBS, and then resuspended with PBS to adjust the cell concentration to 1x10 8 CFU / mL to obtain a bacterial suspension (bacterial body, prepared immediately before use). The freeze-dried bacterial powder was prepared after freeze-drying.

[0026] (3) Mixing of Lactobacillus rhamnosus agent and Sambucus nigra L. ethanol extract (prepared according to the formulation in Table 1)

[0027] Table 1 Different combination ratios of Lactobacillus rhamnosus agent (freeze-dried bacterial powder) and Sambucus nigra L. ethanol extract

[0028] Class Lactobacillus rhamnosus agent Sambucus nigra L. ethanol extract 1 0.5 parts 1 part 2 0.5 parts 5 parts 3 1 part 1 part 4 1 part 5 parts 5 1 part 0.5 parts 6 5 parts 0.5 parts

[0029] (4) Animal test

[0030] One-week adaptive feeding was performed before the formal experiment. The environmental temperature was controlled at 25±1°C, the humidity was controlled at 40%±10%, and the light / dark time was maintained at 12h each. The mice were given sufficient fresh drinking water and basic feed throughout the experiment, and the bedding was changed once a day.

[0031] After the adaptive feeding, the mice were randomly divided into 9 groups: blank control group (Blank), reagent group (Class 1-6), model group (Negtive Control), thymosin positive drug control group (hereinafter referred to as positive group, Thymosin). Among them, the mice in the blank group were only given the same dose of normal saline by gavage throughout the experimental period, and the mice in the reagent group were given 50mg / kg of normal saline by gavage. The gavage amount was 200μL / day. The mice in the thymosin positive control group were given 10mg / kg of thymosin suspension by gavage, and the mice in the model group were given normal saline by gavage. From the 9th day, except for the blank group, the remaining four groups were intraperitoneally injected with 80mg / kg.bw.d of cyclophosphamide (CTX) solution to make an immunosuppression model, for 3 consecutive days. The gavage was maintained during the intraperitoneal injection, and the intraperitoneal injection was stopped from the 12th day of the experiment, and the feeding was continued for 15 consecutive days. The gavage of the mice was started at 10am every day, and the body weight of the mice was weighed and recorded before gavage. Among them, the gavage amount and the intraperitoneal injection amount of CTX were calculated according to the body weight of the mice (0.1mL / 10g). The mice were deprived of food and water 12h before the end of the experiment (the mice were sacrificed). The mice were sacrificed by enucleation and decapitation, and the thymus, spleen, liver and kidney were dissected and weighed accurately.

[0032] The immunocompromised model of mice was established by cyclophosphamide (CTX) induction. By recording the body weight of mice (as shown in FIG. 1), it was found that the combination of the present embodiment could promote the growth and development of mice, and the Class 3 group had the best effect, slowed down the immunosuppressive effect of CTX, and helped the recovery of the body weight of the CTX-induced immunosuppressed mice. CTX caused different degrees of damage to the organs of mice, while the Class 3 of the present embodiment could repair the immunosuppressive damage to the organs caused by CTX, proving that the mixture of Lactobacillus rhamnosus and Sambucus williamsii ethanol extract could effectively protect the immune organs of immunosuppressed mice. Figure 1

[0033] A, organ weight of mice:

[0034] The mice were dissected, the thymus and spleen were washed in physiological saline to remove bloodstains, and then were placed on ice blocks covered with filter paper to dry, and were quickly weighed and recorded. The organ index was calculated according to the following formula:

[0035] Organ index (mg / g) = organ weight (mg) / mouse weight (g)

[0036] As shown in Table 2, the spleen index and thymus index of the negative control group (Negtive Control) mice were slightly lower than those of the blank group, indicating that the cyclophosphamide solution successfully inhibited the development of the immune organs of mice. The spleen index and thymus index of the positive control group (Thymosin) mice were not significantly different from those of the model group. The spleen index and thymus index of the Class 1-6 group mice were increased compared with those of the model group, and the increase of the Class 3 group was the most significant. The results showed that the mixture of Lactobacillus rhamnosus and Sambucus williamsii ethanol extract had the function of immune regulation.

[0037] Table 2 Effect of Lactobacillus rhamnosus agent and Sambucus williamsii ethanol extract mixture on organ index of mice

[0038]

[0039]

[0040] B, determination method of IgM and IgG in serum of mice:

[0041] After the feeding period was completed, quantitative blood was taken from the orbit of the mice, serum was extracted from the blood, and was stored at -20°C. Then, the content of IgM and IgG in the serum was detected by Elisa method.

[0042] ​From Table 3, the IgM and IgG content in serum of Class 1-6 group mice is higher than that of model group (Negtive Control), and the IgM and IgG content of Class 3 group is closer to that of Blank group. It is indicated that the mixture of Lactobacillus rhamnosus and Sambucus williamsii ethanol extract can increase the IgM and IgG content in serum, and the mixture of Lactobacillus rhamnosus and Sambucus williamsii ethanol extract can promote the improvement of immune function of mice in different degrees.

[0043] Table 3 Effect of Lactobacillus rhamnosus agent mixed with Sambucus williamsii ethanol extract on IgM and IgG in serum of mice

[0044] Group IgM (mg / mL) IgG (mg / mL) Blank 290.87±0.67 12.56±0.23 Thymosin 278.98±0.87 11.98±0.65 Negative Control 100.21±0.98 4.76±0.34 Class 1 120.87±0.33 5.98±0.09 Class 2 131.56±0.79 7.09±0.82 Class 3 271.90±0.78 11.97±0.56 Class 4 187.01±0.89 7.99±0.68 Class 5 198.89±0.99 8.56±0.89 Class 6 187.98±0.56 9.08±0.35

[0045] The above merely describes preferred embodiments of the present application, and therefore cannot limit the scope of the present application, that is, equivalent changes and modifications made according to the scope of the present application and the content of the specification should still be within the scope of the present application.

Claims

1. An immune-enhancing composition, characterized in that: It is a mixture of elderberry ethanol extract and Lactobacillus rhamnosus inoculum, wherein, The preparation of elderberry ethanol extract includes: drying elderberries and extracting oil to remove grease, pulverizing them into powder, then refluxing with 95% ethanol, filtering, concentrating and drying to obtain the extract; Lactobacillus rhamnosus inoculant is Lactobacillus rhamnosus Lactobacillus rhamnosus Freeze-dried cells of CICC 6161; The mass ratio of elderberry ethanol extract to Lactobacillus rhamnosus inoculum was 1:

1.

2. The use of the combination of elderberry ethanol extract and Lactobacillus rhamnosus agent in the preparation of an immunomodulatory composition, characterized in that: This combination consists of an elderberry ethanol extract and a Lactobacillus rhamnosus inoculum, in which... The preparation of elderberry ethanol extract includes: drying elderberries and extracting oil to remove grease, pulverizing them into powder, then refluxing with 95% ethanol, filtering, concentrating and drying to obtain the extract; Lactobacillus rhamnosus inoculant is Lactobacillus rhamnosus Lactobacillus rhamnosus Freeze-dried cells of CICC 6161; The mass ratio of elderberry ethanol extract to Lactobacillus rhamnosus inoculum was 1:1.

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