Application of banana fruit extract in immunoregulation

Abstract

The invention provides an application of a plantain fruit extract in immunoregulation, in particular to an application of the plantain fruit extract in regulation of human peripheral blood lymphocyte immunophenotype. Through a multi-target immune regulation mechanism (B cell inhibition, NK function activation and Th polarization), the extract significantly improves the phenotypic imbalance of peripheral blood lymphocytes, and especially shows an outstanding prevention value in people in an immune critical state. The invention provides a new theoretical basis and transformation direction for the development of plant-derived immunomodulators.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and specifically relates to an application of a plantain fruit extract in immune regulation, especially focusing on its regulation effect on the immune phenotype of human peripheral blood lymphocytes, and a new type of immune regulator and immune regulation method developed based on the same. BACKGROUND

[0002] The immune system plays a key role in maintaining human health, and its normal function depends on the coordinated operation of various immune cells. Lymphocytes, as an important component of the immune system, are closely related to the occurrence and development of various diseases due to their abnormal number and function. At present, regulating the proportion and function of lymphocyte subsets has become an important research direction for treating immune-related diseases.

[0003] In existing immune regulation research, although some drugs and methods have been used to regulate the immune system, some of them have problems such as large side effects and unstable efficacy. For example, some immunosuppressants can significantly increase the risk of infection while suppressing immune response, and some immune enhancers can cause excessive immune activation, leading to autoimmune diseases. Therefore, it is of great clinical significance and market demand to develop safe and effective new immune regulators.

[0004] Plantain fruit, as a fruit widely distributed in tropical regions, has been used in traditional medicine for clearing heat and detoxifying, moistening the lungs and relieving cough, etc. Modern research has found that plantain fruit is rich in polyphenols, flavonoids and other bioactive components, and has various pharmacological properties such as antioxidant, anti-inflammatory and anti-tumor. SUMMARY

[0005] Therefore, the present application aims to provide an application of a plantain fruit extract in immune regulation, to further explore the effect of the plantain fruit extract on the immune phenotype of human peripheral blood lymphocytes, to clarify its immune regulation mechanism, and to provide a solid theoretical basis and innovative ideas for the development of new plant-derived immune regulators. At the same time, based on the immune regulation characteristics of the plantain fruit extract, a safe and effective immune regulation method is constructed to improve the immune status of the human body and prevent and assist in the treatment of immune-related diseases.

[0006] To achieve the above-mentioned purpose, the technical scheme of the present application is as follows:

[0007] The application of the plantain fruit extract in the preparation of an immune regulation drug.

[0008] In a specific embodiment provided by the present application, the immune regulation drug is a drug for regulating the immune phenotype of human peripheral blood lymphocytes.

[0009] In one specific embodiment provided in the present application, the immunomodulatory drug is one or more of the drugs for B cell specific inhibition, NK cell function reprogramming and T cell subset reconstruction.

[0010] In one specific embodiment provided in the present application, the plantain fruit extract is obtained by the following method: crushing the stems, leaves, barks or combinations thereof of the plantain family into extraction materials; soaking the extraction materials in a NaCl solution (brine) with a volume percentage of 20-30%, preferably a NaCl solution with a volume percentage of 25%, and heating; extracting the crude extract from the soaked extraction materials by low-temperature wall-breaking method, centrifuging the crude extract, filtering the liquid part and standing in a barrel to become a crude extract solution; and fermenting the crude extract solution with far-infrared stones to obtain the plantain fruit extract.

[0011] In one specific embodiment provided in the present application, the extraction materials are soaked in the NaCl aqueous solution and heated with far-infrared stones, the spectral wavelength of the far-infrared stones is 7.5-9.5 μm, the soaking time is 8-10 h; the crude extract is extracted from the soaked extraction materials by low-temperature wall-breaking method, wherein the low temperature is -40-0℃ and the time is 1-12 h; and the crude extract solution is fermented with far-infrared stones, the spectral wavelength of the far-infrared stone layer is 7.5-9.5 μm.

[0012] Compared with the prior art, the present application has the following advantages:

[0013] The plantain fruit extract is a medicinal and edible material with high safety.

[0014] Precise targeting of immune phenotypes: The plantain fruit extract of the present application is obtained by a unique preparation process, and its active ingredients can specifically act on key subsets of human peripheral blood lymphocytes, achieving synergistic regulation of B cell specific inhibition, NK cell function reprogramming and T cell subset reconstruction. Clinical data show that it can significantly reduce the proportion of CD3+CD19+ mature B cells, increase the proportion of total T cells (CD3+) and helper T cells (CD3+CD4+), optimize the CD4+ / CD8+ ratio, precisely improve the imbalance of lymphocyte phenotypes, and the regulation mechanism is clear and focused on the immune core cell population.

[0015] Subclinical immune disorder intervention advantage is obvious: for immune abnormal population and immune critical state population, the plantain fruit extract of the present application shows differentiated high-efficiency regulation efficiency. Especially for immune critical state population, at least one index normalization rate is 69.23%, and all index complete remission rate is as high as 66.67%, which is significantly better than the improvement effect of immune abnormal population, provides targeted prevention means for the easily ignored health risk stage of immune critical state, and fills the blank of existing immune regulators in the field of subclinical intervention.

[0016] High safety and adaptation to long-term application: the plantain fruit extract of the present application is derived from natural plantain fruit, and after extraction and purification by a specific process, the low toxicity characteristics of plant-derived ingredients are retained. Compared with the side effects such as increased risk of infection and excessive immune activation that may be caused by synthetic immune regulators, the extract regulates immune homeostasis through multi-target synergistic effect, has no obvious adverse reactions, is suitable for long-term use for immune state maintenance and intervention of subclinical immune disorders, and solves the problem of insufficient safety of long-term use of existing immune regulators.

[0017] Overall effect and outstanding response efficiency: the extract has significant regulation effect on multiple immune phenotype indicators, among which the remission rate of abnormal cases of CD3+CD19+ mature B cells is 100%, the remission rates of total T cells (CD3+) and helper T cells (CD3+CD4+) are as high as 71.43% and 77.14% respectively, and NK cells (CD3-CD16+ / CD56+) also show good regulation effect. Its multi-dimensional immune regulation effect can comprehensively improve the number and functional balance of immune cell subpopulations, compared with single-target immune regulators, it can better adapt to the complex synergistic characteristics of the immune system, and the regulation effect is more comprehensive and stable. DETAILED DESCRIPTION

[0018] Unless otherwise defined, the technical terms used in the following examples have the same meanings as generally understood by those skilled in the art to which the present application belongs. The test reagents used in the following examples, unless otherwise specified, are conventional biochemical reagents; the experimental methods, unless otherwise specified, are conventional methods.

[0019] The present application will be described in detail below with reference to the examples.

[0020] The plantain fruit extract is obtained by the following method: crushing the stems, leaves, barks or combinations thereof of the plantain family into the material to be extracted; soaking the material to be extracted in a 25% by volume NaCl aqueous solution and heating; extracting the crude extract from the soaked material to be extracted by low-temperature wall-breaking method, centrifuging the crude extract (3000 rpm for 1 minute), filtering the liquid through a 100 mesh screen and standing for 24 hours to become the crude extract solution; and fermenting the crude extract solution with far infrared stones to obtain the plantain fruit extract. The material to be extracted is soaked in the NaCl aqueous solution and heated with far infrared stones, the spectral wavelength of the far infrared stones is 9.0 μm, and the soaking time is 9 hours; the crude extract is extracted from the soaked material to be extracted by low-temperature wall-breaking method, wherein the low temperature is -20℃ and the time is 10 hours; and the crude extract solution is fermented with far infrared stones for 90 days, the spectral wavelength of the far infrared stone layer is 9 μm.

[0021] The acute oral toxicity test of the plantain fruit extract prepared by the present application is carried out according to GB15193.3-2014 National Food Safety Standard, and the acute oral LD50 of the plantain fruit extract to ICR mice is >5000 mg / kg. According to the acute toxicity classification, it belongs to the actual non-toxic level.

[0022] The subjects of the present application are all volunteers participating in the test and signing the informed consent form. A total of 120 subjects, all cases come from Nanning, Guangxi, time is December 2024, and place is Guangxi Jinyu Medical Maple Blue Coast Station.

[0023] Example 1: Modulatory effect of plantain fruit extract on immune phenotype

[0024] 120 subjects aged 10-80 years (mean ± SD: 45.2 ± 12.8 years) were selected, with a gender ratio of 1:1, and an open-label, single-group, pre-post control trial was conducted. After blood sampling at baseline, the subjects began to take the standardized plantain fruit extract preparation (3g / day, mass concentration of plantain fruit extract preparation 3 ‰) orally every day, and the intervention period was strictly controlled within 20 ± 1 days. Flow cytometry was used to detect peripheral blood lymphocyte subsets, including B lymphocytes (CD3-CD19+), NK cells (CD3-CD16+ or CD56+), T lymphocyte subsets (CD3+ T cells, CD3+CD4+ T cells, CD3+CD8+ T cells, CD3+CD4+ T cells / CD3+CD8+ T cells). The results showed that 5 indicators showed statistically significant differences (P < 0.001) (Table 1, Table 2). Key findings: B cell suppression effect: CD3+CD19+ (B cell marker) decreased significantly (Δ = -0.275%, P < 0.001), suggesting that the intervention may play a role by regulating the humoral immune response. NK cell activity regulation: The proportion of CD3-CD16+ / CD56+ (NK cells) decreased (Δ = -2.430%, P < 0.001), which may be related to the adjustment of immune homeostasis. T cell subset reconstruction: The proportion of total T cells (CD3+) increased significantly (Δ = +3.033%, P < 0.001), helper T cells (CD3+CD4+) increased significantly (Δ = +3.700%, P < 0.001), and the CD4+ / CD8+ ratio improved (Δ = +0.136, P < 0.001), suggesting that the balance of cellular immunity is optimized, indicating that the plantain fruit extract has a significant immunomodulatory effect.

[0025] Table 1

[0026]

[0027] Table 2

[0028]

[0029]

[0030] Example 2: Application of plantain fruit extract in different immune status populations

[0031] The subjects in the immune abnormality group (at least one index is abnormal, which means that CD3+T cell / lymphocyte is not in the range of 50-84, CD3+CD4+T cell / lymphocyte is not in the range of 27-51, CD3+CD8+T cell / lymphocyte is not in the range of 15-44, CD3+CD4+T cell / CD3+CD8+T cell is not in the range of 0.71-2.78, CD3-CD19+cell / lymphocyte is not in the range of 5-18, CD3+CD19+cell / lymphocyte is not in the range of 0-1, CD3-CD16 or CD56+cell / lymphocyte is not in the range of 7-40, and CD3+CD16 or CD56+cell / lymphocyte is not in the range of 0-10) and the critical value group (at least one index is a critical value, which means that the detection value of CD3+T cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (50-84), the detection value of CD3+CD4+T cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (27-51), the detection value of CD3+CD8+T cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (15-44), the detection value of CD3+CD4+T cell / CD3+CD8+T cell is within-10% of the highest value and +10% of the lowest value of the normal value (0.71-2.78), the detection value of CD3-CD19+cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (5-18), the detection value of CD3+CD19+cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (0-1), the detection value of CD3-CD16 or CD56+cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (7-40), and the detection value of CD3+CD16 or CD56+cell / lymphocyte is within-10% of the highest value and +10% of the lowest value of the normal value (0-10)) are intervened respectively. The stratified analysis according to the baseline immune state shows (Tables 3 and 4) that before the intervention, there are 68 people (56.67%) in the immune abnormality group, 39 people (32.50%) in the critical value group, and 13 people (10.83%) in the normal group. After taking the diluted plantain fruit extract (mass concentration 3‰; plantain fruit extract, 1000 ml / day), the proportion of at least one index in the immune abnormality group turning to normal is 52.94% (36 / 68), and the complete remission rate of all indexes is 41.18% (28 / 68); the improvement in the critical value group is more significant, the normalization rate of at least one index is 69.23% (27 / 39), and the complete remission rate is as high as 66.67% (26 / 39), and the comparison between groups has a statistical difference (P=0.008).This result suggests that the plantain fruit extract has better intervention efficiency on subclinical immune disorder population, which may be related to its mechanism of regulating T cell exhaustion state.

[0032] Table 3

[0033]

[0034] Table 4

[0035]

[0036]

[0037] Intervention effect analysis of plantain fruit extract on immune abnormal and critical value population

[0038] The intervention efficiency of the eight immune phenotype indicators CD3-CD19+, CD3+CD19+, CD3-CD16 or CD56+, CD3+CD16 or CD56+, CD3+, CD3+CD4+, CD3+CD8+, and CD3+CD4+ / CD3+CD8+ in the immune abnormal group and the critical value group were 38.236%, 100.000%, 53.846%, 50.000%, 71.428%, 77.142%, 45.833%, and 31.250%, respectively (Table 5). The key findings were that the B cell subpopulation completely responded: all abnormal cases of CD3+CD19+ (mature B cells) were restored (15 / 15), showing absolute effectiveness (OR = ∞, P < 0.001). The T cell subpopulation was dominantly regulated: the remission rates of total T cells (CD3+) and helper T cells (CD3+CD4+) were 71.43% and 77.14%, respectively (P < 0.001). The CD4+ / CD8+ ratio improved limitedly (31.25%), indicating that the intervention was more inclined to Th immune activation rather than balanced regulation. NK / NKT cells responded differently: the remission rate of NK cells (CD3-CD16+ / CD56+) was significantly higher than that of NKT cells (53.85% vs 50.00%, P < 0.05). The critical value population had a high response characteristic: the response rate of the critical value group was significantly higher than that of the immune abnormal group (69.23% vs 52.94%, P = 0.042), indicating that the intervention was more sensitive to the subclinical state regulation.

[0039] Table 5

[0040]

[0041]

[0042] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, and are not intended to limit the present application; although the present application has been described in detail with reference to the above embodiments, those skilled in the art should understand that the technical solutions recorded in the above embodiments can be modified, or some or all of the technical features can be replaced by equivalents; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. Use of Musa sapientum fruit extract in the preparation of an immunomodulatory drug.

2. Use according to claim 1, characterized in that: The immunomodulatory drug is a drug for regulating the immune phenotype of human peripheral blood lymphocytes.

3. Use according to claim 1, characterized in that: The immunomodulatory drug is one or more of the following: B cell-specific inhibition, NK cell function reprogramming, and T cell subset reconstruction.

4. Use according to claim 1, characterized in that: The Musa sapientum fruit extract is obtained by crushing the stems, leaves, barks or combinations thereof of a plant of the Musaceae family into a material to be extracted; soaking the material to be extracted in a NaCl aqueous solution with a volume percentage of 20-30% and heating; extracting the crude extract from the soaked material to be extracted by a low-temperature wall-breaking method, centrifuging the crude extract, filtering the liquid part and standing it in a barrel to become a crude extract liquid; and fermenting the crude extract liquid with far-infrared stones to obtain the Musa sapientum fruit extract.

5. Use according to claim 4, characterized in that: The material to be extracted is soaked in the NaCl aqueous solution and heated with far-infrared stones, the spectral wavelength of the far-infrared stones is 7.5-9.5 μm, and the soaking time is 8-10 h; the crude extract is extracted from the soaked material to be extracted by a low-temperature wall-breaking method, the low temperature is -40-0°C, and the time is 1-12 h; and the crude extract liquid is fermented with far-infrared stones, the spectral wavelength of the far-infrared stone layer is 7.5-9.5 μm.

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