A plant extract composition for enhancing immunity

Abstract

This invention belongs to the field of plant extract technology, specifically relating to a plant extract composition for enhancing immunity. By weight, it is composed of the following raw materials: 3-5 parts Astragalus membranaceus extract, 2.5-4 parts Echinacea purpurea extract, and 1.5-3.5 parts Rhodiola rosea extract. This plant extract composition, through the scientific formulation of Astragalus membranaceus, Echinacea purpurea, and Rhodiola rosea, and an enzymatic extraction process, enhances the immunomodulatory effect. The combination of the three extracts increases the secretion of TNF-α and IL-6 by macrophages. Furthermore, the spleen lymphocyte stimulation index (SI) of the three extracts reaches 5.57, and the lymphocyte transformation rate increases from 191% to 457%. These results indicate that this plant extract composition effectively enhances the body's non-specific and cellular immune functions by activating macrophages and promoting T lymphocyte proliferation through multiple pathways, providing more comprehensive immune protection.

Description

Technical Field

[0001] This invention belongs to the field of plant extract technology, and specifically relates to a plant extract composition for enhancing immunity. Background Technology

[0002] With increasing health awareness, the development of immunomodulatory products using natural plant extracts has become a research hotspot in the pharmaceutical and health product fields. Plant extracts are naturally derived, highly safe, and rich in various immunomodulatory components such as polysaccharides, flavonoids, and saponins. Astragalus, Echinacea, and Rhodiola are commonly used immunomodulatory plant raw materials: Astragalus and Echinacea extracts can regulate immune cell activity and promote the secretion of immune factors; Rhodiola extract can improve the body's immune tolerance. The preparation of plant extracts typically employs processes such as enzymatic hydrolysis, ultrasound-assisted extraction, hot water extraction, vacuum concentration, and spray drying. These processes extract, purify, and dry the active ingredients from plants to obtain extracts that can be used for subsequent product preparation.

[0003] However, most products use a single plant extract as an immune-modulating ingredient, resulting in a relatively singular effect and making it difficult to achieve a comprehensive immune-boosting effect. Although some studies have combined multiple extracts, the ratios are mostly based on empirical design and have not been scientifically screened and optimized, making it difficult to leverage the synergistic effects between different components, and may even lead to antagonism due to improper ratios.

[0004] The information disclosed in this background section is intended only to enhance the understanding of the overall background of the invention and should not be construed as an admission or in any way implying that the information constitutes prior art known to those skilled in the art. Summary of the Invention

[0005] The purpose of this invention is to provide a plant extract composition that enhances immunity, thereby addressing the problems existing in the prior art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: An immune-boosting plant extract composition, by weight, is composed of the following ingredients: 3-5 parts Astragalus membranaceus extract, 2.5-4 parts Echinacea purpurea extract, and 1.5-3.5 parts Rhodiola rosea extract.

[0007] Furthermore, the preparation method of the Astragalus extract is as follows: The dried roots of Astragalus membranaceus are washed, dried, and pulverized through a 40-mesh sieve. Purified water is added at a material-to-liquid ratio of 1g:15mL, and the pH of the system is adjusted to 4.8-5.0. Cellulase at 1.2% of the mass of the dried Astragalus membranaceus powder is added to the system, and enzymatic hydrolysis is performed at 50±2℃ for 90min, followed by enzyme inactivation at elevated temperature. Then, ultrasonic extraction is performed at 450W and 20kHz for 35min. The extract is centrifuged at 5000r / min for 15min, coarsely filtered through a 200-mesh sieve, and finely filtered through a 0.45μm filter membrane. The filtrate is collected, concentrated under reduced pressure using a rotary evaporator to a solid content of 45-50%, and then spray-dried to obtain the Astragalus extract.

[0008] Further, the preparation method of the echinacea extract is as follows: the dried roots of echinacea are washed and dried, pulverized and passed through a 40-mesh sieve, purified water is added at a material-to-liquid ratio of 1g:15mL, and the pH of the system is adjusted to 4.8-5.0; 0.8% (by weight of echinacea powder) of cellulase is added to the system, and enzymatic hydrolysis is carried out at 50±2℃ for 90min, and then the enzyme is inactivated by heating; then 6 times the mass of purified water is added and stirred at 70±5℃ and 100r / min for 60min; the extract is centrifuged at 5000r / min for 15min, coarsely filtered through a 200-mesh sieve and finely filtered through a 0.45μm filter membrane, and the filtrate is collected; 3 times the volume of 95% ethanol is added to the filtrate and precipitated overnight at 4℃; the precipitate is concentrated under reduced pressure in a rotary evaporator to a solid content of 45-50% and then spray-dried to obtain the echinacea extract.

[0009] Furthermore, the preparation method of the Rhodiola rosea extract is as follows: the dried rhizomes of Rhodiola rosea are washed and dried, pulverized and passed through a 30-mesh sieve, and purified water is added at a material-to-liquid ratio of 1g:12mL, and the pH of the system is adjusted to 6.8-7.2; 0.8% by weight of neutral protease of Rhodiola rosea powder is added to the system, and enzymatic hydrolysis is carried out at 45±2℃ for 90min, and then the enzyme is inactivated by heating; then 8 times the amount of purified water is added and stirred and extracted at 70±5℃ and 100r / min for 90min; the extract is centrifuged at 8000r / min for 10min and filtered through a 0.45μm filter membrane, and the filtrate is collected; the filtrate is concentrated under reduced pressure in a rotary evaporator to a solid content of 25-30% and then spray-dried to obtain the Rhodiola rosea extract.

[0010] Furthermore, the mass ratio of the Astragalus extract, Echinacea extract, and Rhodiola rosea extract is 4:3:2.

[0011] Compared with the prior art, the present invention has the following beneficial effects: The plant extract composition presented in this application, through the scientific formulation of Astragalus membranaceus, Echinacea purpurea, and Rhodiola rosea, and an enzymatic extraction process, enhances the immunomodulatory effect. The combination of the three extracts increased the secretion of TNF-α and IL-6 by macrophages. Furthermore, the spleen lymphocyte stimulation index (SI) of the three extracts reached 5.57, and the lymphocyte transformation rate increased from 191% to 457%. These results indicate that the plant extract composition of this application effectively enhances the body's non-specific and cellular immune functions by activating macrophages and promoting T lymphocyte proliferation through multiple pathways, providing more comprehensive immune protection. Detailed Implementation

[0012] The technical solution of this invention patent will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention.

[0013] 1. Preparation of Astragalus extract After washing, the dried roots of Astragalus membranaceus were dried in a forced-air dryer at 50℃ until the moisture content was ≤8%. The powder was then pulverized and passed through a 40-mesh sieve. Purified water was added at a material-to-liquid ratio of 1g:15mL, and the pH was adjusted to 5.0 with disodium hydrogen phosphate-citric acid buffer. Cellulase (enzyme activity 50000U / g) was then added for enzymatic hydrolysis at 1.2% of the Astragalus membranaceus powder. Hydrolysis was carried out at 50±2℃ for 90min. After hydrolysis, the temperature was raised to 95℃ and held for 10min to inactivate the enzyme. Then, 450W... The extract was ultrasonically extracted at 20 kHz and 40 ± 3 ℃ for 35 min, and the extraction was repeated twice and the extracts were combined. The extract was centrifuged at 5000 r / min for 15 min, coarsely filtered through a 200-mesh sieve and finely filtered through a 0.45 μm filter membrane, and the filtrate was collected. The filtrate was concentrated under reduced pressure (55 ± 5 ℃, vacuum degree -0.085 MPa) in a rotary evaporator until the solid content was 50%, and then spray-dried (inlet air temperature 175 ± 5 ℃, outlet air temperature 85 ± 5 ℃) to obtain the Astragalus extract.

[0014] 2. Preparation of Echinacea extract After cleaning and removing impurities, the dried roots of *Echinacea purpurea* were dried in a forced-air drying process at 50℃ until the moisture content was ≤8%. The roots were then pulverized and passed through a 40-mesh sieve. Purified water was added at a liquid ratio of 11g:15mL, and the pH was adjusted to 5.0 with disodium hydrogen phosphate-citric acid buffer. Cellulase (enzyme activity 50000U / g) was then added for enzymatic hydrolysis at 0.8% of the *Echinacea purpurea* powder. Hydrolysis was carried out at 50±2℃ for 90 min. After hydrolysis, the temperature was raised to 95℃ and incubated for 10 min to inactivate the enzyme. Six times the mass of purified water was added, and the mixture was incubated at 70±5℃ for 100 minutes. The mixture was stirred and extracted for 60 min at 5000 r / min, and the extraction was repeated twice and the extracts were combined. The extract was centrifuged at 5000 r / min for 15 min, coarsely filtered through a 200-mesh sieve and finely filtered through a 0.45 μm filter membrane, and the filtrate was collected. Three volumes of 95% ethanol were added to the filtrate and the precipitate was allowed to precipitate overnight at 4 °C. The precipitate was concentrated under reduced pressure (55±5 °C, vacuum degree -0.085 MPa) using a rotary evaporator until the solid content was 45%, and then spray-dried (inlet air temperature 170±5 °C, outlet air temperature 80±5 °C) to obtain the echinacea extract.

[0015] 3. Preparation of Rhodiola Rosea Extract After cleaning and removing impurities, the dried rhizomes of Rhodiola rosea were dried in a forced-air drying process at 50℃ until the moisture content was ≤8%. The dried rhizomes were then pulverized and passed through a 30-mesh sieve. Purified water was added at a material-to-liquid ratio of 1g:12mL, and the pH was adjusted to 7.0 with phosphate buffer. Then, neutral protease (enzyme activity 40000U / g) was added for enzymatic hydrolysis at 0.8% of the Rhodiola rosea powder mass. The hydrolysis was carried out at 45±2℃ for 90 minutes, followed by inactivation at 90℃ for 10 minutes. Add 8 times the volume of purified water and stir at 70±5℃ and 100r / min for 90min. Repeat the extraction twice and combine the extracts. Centrifuge the extract at 8000r / min for 10min and filter through a 0.45μm membrane. Collect the filtrate and concentrate it under reduced pressure (55±5℃, vacuum degree -0.08MPa) using a rotary evaporator until the solid content is 30%. Spray dry (inlet air temperature 170±5℃, outlet air temperature 80±5℃) to obtain the Rhodiola rosea extract.

[0016] 4. Effects of different extracts on macrophages RAW264.7 cells were cultured in RPMI-1640 containing 10% fetal bovine serum (FBS) at 37°C and 5% CO2. Cells were passaged when the confluence reached 80-90%.

[0017] RAW264.7 cells in logarithmic growth phase were harvested and treated with 1×10⁻⁶ cells. 5Cells were seeded at a density of 100 μL / mL in 96-well plates and cultured at 37°C and 5% CO2 for 24 h. The supernatant was discarded, and serum-free RPMI-1640 medium containing the test substance (final concentration 100 μg / mL) was added. A blank control group was also included. Each group had three replicates. After another 24 h of culture, the cell supernatant was collected. Cell viability was assessed using the MTT assay (absorbance measured at 490 nm). TNF-α and IL-6 secretion levels were measured using ELISA. A cell viability ≥90% was considered to indicate no cytotoxicity, and significantly higher levels of TNF-α and IL-6 compared to the blank group were considered to indicate effective immune activation. Results are shown in Table 1.

[0018] Table 1. Activation effects of different extracts on macrophages

[0019] As shown in Table 1, all three extracts, when used individually, can promote the secretion of TNF-α and IL-6 by macrophages, effectively activating macrophages, which is an important pathway to enhance the body's non-specific immunity. When any two extracts are combined, the secretion of TNF-α and IL-6 is significantly higher than that of the single components. When the three extracts are combined in a 1:1:1 ratio, the secretion of TNF-α reaches 305.2 pg / mL and the secretion of IL-6 reaches 385.6 pg / mL. When the three are combined in a 4:3:2 ratio, the effect is further enhanced, indicating that Astragalus membranaceus, Echinacea purpurea, and Rhodiola rosea can synergistically activate macrophages from different immune pathways, forming a more comprehensive non-specific immune enhancement effect.

[0020] 5. Effects of different extracts on cellular immune function Clean-grade ICR mice (18-22g) were randomly divided into multiple groups of 10 mice each (half male and half female) according to body weight. After acclimatization for 7 days, the test substance (equivalent to 5g / kg / day of raw drug) was administered by gavage daily at a volume of 0.2mL / 10g body weight for 30 consecutive days. Twenty-four hours after the last administration, the mice were euthanized by cervical dislocation. The spleen was aseptically removed and placed in a petri dish containing RPMI-1640 incomplete medium. Single-cell suspensions were obtained by squeezing the spleen with a sterile glass slide, filtered through a 200-mesh sieve, centrifuged at 1000 rpm for 5 min, and the supernatant was discarded. 3 mL of erythrocyte lysis buffer (NH4Cl 0.155mol / L) was added, and the mixture was allowed to stand for 2 min, then centrifuged at 1000 rpm for 5 min. The mice were washed twice with RPMI-1640 complete medium, and the concentration was adjusted to 5×10⁻⁶. 8 cells / mL; add 100 μL (5 × 10⁶ cells / mL) of spleen cell suspension to each well of a 96-well plate. 5Cells were used in experimental wells (cells + concanavalin A final concentration 5 μg / mL) and control wells (cells only). Each group had 3 replicates. After incubation at 37℃ and 5% CO2 for 44 h, 10 μL of MTT was added to each well, and the cells were incubated for another 4 h. After centrifugation at 1000 r / min for 5 min, the supernatant was discarded, and 150 μL of DMSO was added to each well. The OD value was measured at 570 nm using a microplate reader. The stimulation index SI was calculated as OD value of experimental wells / OD value of control wells. An SI > 1 was considered to promote lymphocyte proliferation. The higher the SI, the stronger the enhancement of cellular immune function by the test substance. The lymphocyte transformation rate (%) was calculated as (SI - 1) × 100%. The results are shown in Table 2.

[0021] Table 2. Effects of different extracts on cellular immune function

[0022] Table 2 shows that all three extracts, when used alone, significantly promoted ConA-induced splenic lymphocyte proliferation. Enhanced T lymphocyte proliferation directly reflects improved cellular immune function, contributing to enhanced antiviral and antitumor capabilities. Combinations of any two extracts resulted in higher lymphocyte proliferation rates than single-component combinations: the SI for the Astragalus and Echinacea combination (a+b) reached 5.08, the SI for the Astragalus and Rhodiola combination (a+c) reached 5.13, and the SI for the Echinacea and Rhodiola combination (b+c) reached 5.00. The SI for the three extracts in a 1:1:1 ratio reached 5.56, and the SI for the 4:3:2 ratio reached 5.57. The lymphocyte transformation rate increased from 191% in the control group to 457%, indicating that the three-component combination maximized T lymphocyte activation and established a robust cellular immune response.

[0023] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A plant extract composition for enhancing immunity, characterized in that, It is composed of the following ingredients in parts by weight: 3-5 parts Astragalus membranaceus extract, 2.5-4 parts Echinacea purpurea extract and 1.5-3.5 parts Rhodiola rosea extract.

2. The plant extract composition for enhancing immunity according to claim 1, characterized in that, The preparation method of the Astragalus extract is as follows: The dried roots of Astragalus membranaceus are washed, dried, and pulverized through a 40-mesh sieve. Purified water is added at a ratio of 1g:15mL, and the pH of the system is adjusted to 4.8-5.

0. Cellulase at 1.2% of the mass of the dried Astragalus membranaceus powder is added to the system, and enzymatic hydrolysis is performed at 50±2℃ for 90 min, followed by enzyme inactivation at elevated temperature. Then, ultrasonic extraction is performed at 450W and 20kHz for 35 min. The extract is centrifuged at 5000r / min for 15 min, coarsely filtered through a 200-mesh sieve, and finely filtered through a 0.45μm filter membrane. The filtrate is collected, concentrated under reduced pressure using a rotary evaporator to a solid content of 45-50%, and then spray-dried to obtain the Astragalus extract.

3. The plant extract composition for enhancing immunity according to claim 1, characterized in that, The preparation method of the echinacea extract is as follows: the dried roots of echinacea are washed and dried, pulverized and passed through a 40-mesh sieve, purified water is added at a material-to-liquid ratio of 1g:15mL, and the pH of the system is adjusted to 4.8-5.0; 0.8% (by weight of echinacea powder) of cellulase is added to the system, and enzymatic hydrolysis is carried out at 50±2℃ for 90min, followed by enzyme inactivation by heating; then 6 times the mass of purified water is added and the mixture is stirred and extracted at 70±5℃ and 100r / min for 60min; the extract is centrifuged at 5000r / min for 15min, coarsely filtered through a 200-mesh sieve and finely filtered through a 0.45μm filter membrane, and the filtrate is collected; 3 times the volume of 95% ethanol is added to the filtrate and precipitated overnight at 4℃; the precipitate is concentrated under reduced pressure in a rotary evaporator to a solid content of 45-50% and then spray-dried to obtain the echinacea extract.

4. The plant extract composition for enhancing immunity according to claim 1, characterized in that, The preparation method of the Rhodiola rosea extract is as follows: The dried rhizomes of Rhodiola rosea from the mountains are washed, dried, pulverized, and passed through a 30-mesh sieve. Purified water is added at a material-to-liquid ratio of 1g:12mL, and the pH of the system is adjusted to 6.8-7.

2. Neutral protease at 0.8% of the mass of the Rhodiola rosea powder is added to the system, and enzymatic hydrolysis is performed at 45±2℃ for 90min, followed by enzyme inactivation by heating. Then, 8 times the volume of purified water is added, and the mixture is stirred and extracted at 70±5℃ and 100r / min for 90min. The extract is centrifuged at 8000r / min for 10min and filtered through a 0.45μm filter membrane. The filtrate is collected. The filtrate is concentrated under reduced pressure using a rotary evaporator to a solid content of 25-30%, and then spray-dried to obtain the Rhodiola rosea extract.

5. The plant extract composition for enhancing immunity according to claim 1, characterized in that, The mass ratio of Astragalus membranaceus extract, Echinacea purpurea extract and Rhodiola rosea extract is 4:3:2.

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