A traditional Chinese medicine nebulizer liquid for improving immunity and a preparation method thereof

Abstract

The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine atomization liquid for improving immunity and a preparation method thereof. The traditional Chinese medicine atomization liquid for improving immunity comprises the following raw materials in parts by weight: 23-28 parts of rhizoma polygonati, 15-18 parts of cornu cervi, 10-14 parts of fructus meliae toosendan, 9-13 parts of lycium chinense, 11-14 parts of maca, 7-11 parts of rhizoma atractylodis, 3-5 parts of no tail fruit, 5-8 parts of semen ziziphi spinosae, 9-11 parts of radix ophiopogonis, 6-10 parts of poria cocos and 1-3 parts of radix glycyrrhizae. The whole prescription is tonifying but not stagnating, and is warm but not dry, and can systematically improve the immune function of the body and relieve fatigue, so as to achieve the effect of treating the symptoms and the root cause. The application has high safety and mildness, and can be quickly absorbed after atomization administration, has high drug utilization rate, and has rapid drug effect.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine nebulizer for enhancing immunity and its preparation method. Background Technology

[0002] Immunity is a vital physiological barrier for the body to resist the invasion of pathogenic microorganisms and maintain homeostasis. With the fast pace of modern life, increased work pressure, and irregular eating and sleeping habits, the number of people in a sub-healthy state is rising, and fatigue and weakened immunity have become the most common health problems. A weakened immune system makes the body more susceptible to pathogens, inducing various diseases and seriously endangering human health. Therefore, strengthening the body's immune function is key to preventing disease and promoting patient recovery.

[0003] Currently, while chemical drugs on the market that boost immunity can alleviate fatigue and enhance immune function to some extent, most are cortical stimulants, and long-term use can easily lead to addiction and significant toxic side effects. Traditional Chinese medicine (TCM) has a long history and unique advantages in regulating the body's immune function, emphasizing mobilizing the body's own immunity and focusing on holistic treatment that addresses both the symptoms and the root cause. However, most existing TCM preparations are currently oral formulations or granules, which undergo digestion and decomposition in the gastrointestinal tract and first-pass metabolism in the liver, resulting in significant loss of effective components, low bioavailability, and slow onset of action.

[0004] Nebulized inhalation, as a novel route of drug delivery, allows medications to be rapidly absorbed into the bloodstream through the respiratory mucosa and pulmonary capillaries after nebulization. It offers advantages such as rapid onset of action, avoidance of the first-pass effect, and high drug utilization. Existing research indicates that nebulized inhalation of traditional Chinese medicine (TCM) has potential in regulating the body's immune function. However, there is still a lack of TCM nebulization products specifically designed to enhance immunity, and existing products often suffer from problems such as simple components, limited efficacy, and inadequate formulation. For example, many publicly available compound TCM nebulization preparations for enhancing immunity primarily use extracts of TCM polysaccharides and polypeptides as main components, requiring complex chemical modification processes that are cumbersome and result in significant loss of effective ingredients. While some herbal nebulization compositions contain herbs such as Polygonatum sibiricum, their formulation design lacks systematic guidance from TCM theory, and the compatibility of the principal, assistant, and adjuvant herbs is not clearly defined.

[0005] Therefore, there is an urgent need to develop a traditional Chinese medicine nebulization product that is rationally formulated, safe, effective, and easy to prepare to enhance immunity. Summary of the Invention

[0006] The primary objective of this invention is to provide a traditional Chinese medicine nebulizer solution that enhances immunity.

[0007] The second objective of this invention is to provide a method for preparing a traditional Chinese medicine nebulizer solution to enhance immunity.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A traditional Chinese medicine nebulizer for enhancing immunity comprises the following raw materials in parts by weight: 23-28 parts Polygonatum sibiricum, 15-18 parts deer antler, 10-14 parts of Cynanchum paniculatum fruit, 9-13 parts of wolfberry, 11-14 parts of maca, 7-11 parts of Atractylodes macrocephala, 3-5 parts of aralia elata fruit, 5-8 parts of Ziziphus jujuba seed, 9-11 parts of Ophiopogon japonicus, 6-10 parts of Poria cocos, and 1-3 parts of licorice.

[0009] Furthermore, a traditional Chinese medicine nebulizer for enhancing immunity includes the following ingredients in parts by weight: 26 parts Polygonatum sibiricum, 17 parts deer antler, 11 parts Cycas revoluta fruit, 10 parts wolfberry, 13 parts maca, 8 parts Atractylodes macrocephala, 4 parts jujube seed, 6 parts Ophiopogon japonicus, 10 parts Poria cocos, and 3 parts licorice.

[0010] A method for preparing a traditional Chinese medicine nebulizer to enhance immunity includes the following steps: (1) Take each raw material according to the weight, remove impurities, clean, crush, add water to soak, decoct twice, collect the decoction, filter, and combine the decoctions; (2) Concentrate the medicinal liquid obtained in step (1) to obtain a concentrated liquid, then add ethanol to the concentrated liquid and let it stand for 24 hours, filter, concentrate the filtrate to obtain an extract, then add water to the extract to obtain a traditional Chinese medicine solution, mix evenly, filter, sterilize, and dispense to obtain a traditional Chinese medicine atomizing liquid.

[0011] Furthermore, in step (1), the particle size of each raw material after pulverization is 50-80 mesh.

[0012] Furthermore, the amount of water added is 10-13 times the total mass of each raw material.

[0013] Furthermore, the soaking time in step (1) is 20-30 minutes.

[0014] Furthermore, the temperature of the decoction in step (1) is 90-100℃ and the time is 25-30min.

[0015] Further, the relative density of the concentrate in step (2) is 1.0-1.2; the volume of the ethanol accounts for 50-60% of the volume of the concentrate; and the relative density of the extract is 1.1-1.3.

[0016] Furthermore, in step (2), the ratio of extract to water in the herbal solution is (3-5) g: 100 mL.

[0017] The efficacy of each Chinese herb in the formula is analyzed as follows: Polygonatum: [Properties and Channels Entered] Sweet, neutral. Enters the spleen, lung, and kidney channels. [Functions and Indications] Tonifies qi and nourishes yin, strengthens the spleen, moistens the lungs, and benefits the kidneys. Used for spleen and stomach qi deficiency, fatigue, stomach yin deficiency, dry mouth and poor appetite, lung deficiency and dry cough, consumptive cough with hemoptysis, deficiency of essence and blood, soreness and weakness of the lower back and knees, premature graying of hair, and internal heat and thirst.

[0018] Deer antler: [Properties and Channels Entered] Sweet, salty, warm. Enters the kidney and liver channels. [Functions and Indications] Strengthens kidney yang, nourishes essence and blood, strengthens tendons and bones, regulates the Chong and Ren meridians, and promotes the healing of sores and carbuncles. Used for kidney yang deficiency, deficiency of essence and blood, impotence and premature ejaculation, infertility due to cold uterus, emaciation, fatigue, aversion to cold, dizziness, tinnitus, deafness, cold pain in the lower back and spine, weakness of tendons and bones, metrorrhagia and leukorrhea, and non-healing carbuncles.

[0019] Fragrant Tree Fruit: [Properties and Channels Entered] Pungent; warm in nature. Enters the Lung, Spleen, and Large Intestine channels. [Functions and Indications] Promotes blood circulation and qi flow, relieves pain, kills parasites and relieves itching. Used for internal injuries causing chest and abdominal distension and pain, traumatic injuries, muscle and bone pain, carbuncles, eczema, and scabies.

[0020] Goji berries: [Properties and Channels Entered] Sweet, neutral. Enters the Liver and Kidney channels. [Functions and Indications] Nourishes the Liver and Kidneys, benefits essence and improves eyesight. Used for deficiency of essence and blood, soreness and weakness of the lower back and knees, dizziness and tinnitus, impotence and seminal emission, internal heat and thirst, blood deficiency and chlorosis, and blurred vision.

[0021] Atractylodes macrocephala: [Properties and Channels Entered] Bitter, sweet, warm. Enters the spleen and stomach channels. [Functions and Indications] Strengthens the spleen and replenishes qi, dries dampness and promotes diuresis, stops sweating, and calms the fetus. Used for spleen deficiency with poor appetite, abdominal distension and diarrhea, phlegm retention with dizziness and palpitations, edema, spontaneous sweating, and threatened abortion.

[0022] Tailless fruit: [Properties and Channels Entered] Bitter and cool in nature. Enters the Liver and Kidney channels. [Functions and Indications] Calms the liver and extinguishes wind, clears heat and detoxifies. Primarily used to treat hypertension and hepatitis.

[0023] Sour jujube seed: [Properties and Channels Entered] Sweet, sour, neutral. Enters the liver, gallbladder, and heart channels. [Functions and Indications] Nourishes the heart and liver, calms the mind and soothes the nerves, astringes sweat, and generates body fluids. Used for insomnia due to deficiency and restlessness, palpitations and excessive dreaming, excessive sweating due to physical weakness, and thirst due to fluid depletion.

[0024] Ophiopogon japonicus: [Properties and Channels Entered] Sweet, slightly bitter, slightly cold. Enters the Heart, Lung, and Stomach channels. [Functions and Indications] Nourishes Yin and generates fluids, moistens the lungs and clears the heart. Used for dry cough due to lung dryness, consumptive cough due to Yin deficiency, sore throat, thirst due to fluid depletion, internal heat and thirst, irritability and insomnia, and constipation due to intestinal dryness.

[0025] Poria cocos: [Properties and Channels Entered] Sweet, bland, neutral. Enters the Heart, Lung, Spleen, and Kidney channels. [Functions and Indications] Promotes diuresis and eliminates dampness, strengthens the spleen, and calms the mind. Used for edema with scanty urine, phlegm retention with dizziness and palpitations, spleen deficiency with poor appetite, loose stools and diarrhea, restlessness, palpitations, and insomnia.

[0026] Licorice: [Properties and Channels Entered] Sweet, neutral; enters the Heart, Lung, Spleen, and Stomach channels. [Functions and Indications] Tonifies the spleen and replenishes qi, clears heat and detoxifies, resolves phlegm and relieves cough, alleviates spasms and pain, and harmonizes the effects of other herbs. Used for spleen and stomach weakness, fatigue, palpitations, shortness of breath, cough with excessive phlegm, abdominal and limb spasms and pain, carbuncles and boils, and to alleviate the toxicity and harshness of other drugs.

[0027] Maca: Native to the Andes Mountains of South America. It is rich in amino acids, minerals, fatty acids, vitamins, and various active substances. Its alkaloid components have antibacterial, antitumor, anemia-improving, and cardiovascular disease-preventing effects.

[0028] Compared with the prior art, the main advantages of the present invention are as follows: This invention provides a traditional Chinese medicine nebulizer solution for enhancing immunity and its preparation method. Following the TCM theory of "principal, assistant, adjuvant, and guide," the principal herbs are Polygonatum sibiricum, deer antler, and Cynanchum paniculatum fruit. Polygonatum sibiricum nourishes the kidneys and yin, deer antler benefits essence and assists yang, and Cynanchum paniculatum fruit invigorates blood and promotes qi circulation. These three herbs work synergistically to nourish qi, blood, yin, and yang, fundamentally replenishing the body's vital energy and enhancing immunity. The assistant herbs are Lycium barbarum, maca, Atractylodes macrocephala, and Prunus armeniaca. Lycium barbarum nourishes the liver and kidneys, maca tonifies the kidneys and strengthens yang to support the body's foundation, Atractylodes macrocephala invigorates the spleen and benefits qi, and Prunus armeniaca promotes qi circulation and relieves pain to regulate the middle jiao's qi mechanism. The adjuvant herbs are Ziziphus jujuba seed to nourish the heart and liver, Ophiopogon japonicus to benefit the stomach and generate fluids to calm the mind and moisten dryness, and Poria cocos to calm the mind and improve heart and spirit malnutrition and sleep deficiency. Glycyrrhiza uralensis harmonizes all the herbs, ensuring the entire formula is nourishing without stagnation and warming without dryness. This systemically enhances the body's immune function while also relieving fatigue, achieving a comprehensive treatment effect. The formula of this invention is mild and safe, and can be rapidly absorbed through nebulization, resulting in high drug utilization and rapid onset of action. Detailed Implementation

[0029] The technical solution of the present invention will be further described below with reference to specific embodiments. However, those skilled in the art should understand that the following embodiments are only for illustrating the present invention and should not be regarded as limiting the present invention. Specific conditions not specified in the embodiments are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the reagents or instruments used are all conventional products obtained through commercial channels.

[0030] Example 1 A traditional Chinese medicine nebulizer for enhancing immunity, comprising the following raw materials in parts by weight: 26 parts Polygonatum sibiricum, 17 parts deer antler, 11 parts Cycas revoluta fruit, 10 parts wolfberry, 13 parts maca, 8 parts Atractylodes macrocephala, 4 parts jujube seed, 6 parts Ophiopogon japonicus, 10 parts Poria cocos, and 3 parts licorice.

[0031] A method for preparing a traditional Chinese medicine nebulizer to enhance immunity includes the following steps: (1) Take each raw material according to the above weight proportions, remove impurities, clean, crush, pass through a 60-mesh sieve, add water with a weight of 12 times the total weight of the raw materials and soak for 25 minutes, then decoct at 95℃ for 30 minutes, decoct twice, collect the decoction, filter, and combine the decoctions. (2) Concentrate the medicinal liquid obtained in step (1) into a concentrate with a relative density of 1.1, then add ethanol to the concentrate (to make the ethanol content reach 55% v / v) and let it stand for 24 hours to remove macromolecular impurities and reduce the irritation during atomization. Filter and concentrate the filtrate until there is no ethanol smell to obtain an extract with a relative density of 1.2. Then add water to the extract, with the ratio of extract to water being 4g:100mL to obtain a traditional Chinese medicine solution. Mix evenly, filter, sterilize, and dispense to obtain the traditional Chinese medicine atomization solution.

[0032] Example 2 A traditional Chinese medicine nebulizer for enhancing immunity, comprising the following raw materials in parts by weight: 23 parts Polygonatum sibiricum, 15 parts deer antler, 10 parts Cycas revoluta fruit, 9 parts wolfberry, 11 parts maca, 7 parts Atractylodes macrocephala, 3 parts jujube seed, 5 parts Ophiopogon japonicus, 9 parts Poria cocos, and 1 part licorice.

[0033] A method for preparing a traditional Chinese medicine nebulizer to enhance immunity includes the following steps: (1) Take each raw material according to the above weight parts, remove impurities, clean, crush, pass through a 50-mesh sieve, add water with a weight of 10 times the total weight of the raw materials and soak for 20 minutes, then decoct at 90℃ for 30 minutes, decoct twice, collect the decoction, filter, and combine the decoctions; (2) Concentrate the medicinal liquid obtained in step (1) into a concentrate with a relative density of 1.0, then add ethanol to the concentrate (to make the ethanol content reach 50% v / v) and let it stand for 24 hours to remove macromolecular impurities and reduce the irritation during atomization. Filter and concentrate the filtrate until there is no ethanol smell to obtain an extract with a relative density of 1.1. Then add water to the extract, with the ratio of extract to water being 3g:100mL to obtain a traditional Chinese medicine solution. Mix evenly, filter, sterilize, and dispense to obtain the traditional Chinese medicine atomization solution.

[0034] Example 3 A traditional Chinese medicine nebulizer for enhancing immunity, comprising the following raw materials in parts by weight: 28 parts Polygonatum sibiricum, 18 parts deer antler, 14 parts Cycas revoluta fruit, 13 parts wolfberry, 14 parts maca, 11 parts Atractylodes macrocephala, 5 parts aralia elata, 8 parts Ziziphus jujuba seed, 11 parts Ophiopogon japonicus, 10 parts Poria cocos, and 3 parts licorice.

[0035] A method for preparing a traditional Chinese medicine nebulizer to enhance immunity includes the following steps: (1) Take each raw material according to the above weight proportions, remove impurities, clean, crush, pass through an 80-mesh sieve, add water with a weight of 13 times the total weight of the raw materials and soak for 30 minutes, then decoct at 100℃ for 25 minutes, decoct twice, collect the decoction, filter, and combine the decoctions. (2) Concentrate the medicinal liquid obtained in step (1) into a concentrate with a relative density of 1.2, then add ethanol to the concentrate (to make the ethanol content reach 60% v / v) and let it stand for 24 hours to remove macromolecular impurities and reduce the irritation during atomization. Filter and concentrate the filtrate until there is no ethanol smell to obtain an extract with a relative density of 1.3. Then add water to the extract, with the ratio of extract to water being 5g:100mL to obtain a traditional Chinese medicine solution. Mix evenly, filter, sterilize, and dispense to obtain the traditional Chinese medicine atomization solution.

[0036] Comparative Example 1 A traditional Chinese medicine nebulizer solution for enhancing immunity comprises the following ingredients in parts by weight: 26 parts Polygonatum sibiricum, 17 parts deer antler, 10 parts wolfberry, 13 parts maca, 8 parts Atractylodes macrocephala, 4 parts aralia elata, 6 parts Ziziphus jujuba var. spinosa, 10 parts Ophiopogon japonicus, 7 parts Poria cocos, and 3 parts licorice. Specifically, the fruit of the fragrant tree is omitted from the formula. All other aspects remain consistent with Example 1.

[0037] Comparative Example 2 A traditional Chinese medicine nebulizer solution for enhancing immunity, comprising the following ingredients in parts by weight: 26 parts Polygonatum sibiricum, 17 parts deer antler, 11 parts Cycas revoluta fruit, 10 parts Lycium barbarum, 13 parts Maca, 8 parts Atractylodes macrocephala, 6 parts Ziziphus jujuba seed, 10 parts Ophiopogon japonicus, 7 parts Poria cocos, and 3 parts Glycyrrhiza uralensis. Specifically, the tailless fruit is omitted from the formula. All other aspects remain consistent with Example 1.

[0038] Comparative Example 3 A traditional Chinese medicine nebulizer solution for enhancing immunity comprises the following ingredients in parts by weight: 26 parts Polygonatum sibiricum, 17 parts deer antler, 11 parts Salvia miltiorrhiza, 10 parts Lycium barbarum, 13 parts maca, 8 parts Atractylodes macrocephala, 4 parts Citrus aurantium, 6 parts Ziziphus jujuba var. spinosa, 10 parts Ophiopogon japonicus, 7 parts Poria cocos, and 3 parts Glycyrrhiza uralensis. Specifically, Salvia miltiorrhiza is used instead of Citrus aurantium fruit in the formula. Other aspects remain consistent with Example 1.

[0039] Pharmacodynamics test 1. Experimental Animals: SPF-grade ICR male mice, 20-25g each, were housed in a clean environment with 12-hour alternating light and dark lighting, a constant temperature of 25℃, and relative humidity of 60%. Sufficient drinking water and feed were provided, and cage bedding was changed daily. Animals were allowed one week to acclimatize to the environment before the experiment.

[0040] 2. Experimental grouping and construction of immunosuppression model ICR mice were then randomly divided into 10 groups, with 15 mice in each group. These were: control group, model group, low-dose group (Example 1), medium-dose group (Example 1), high-dose group (Example 1), low-dose group (Example 2), low-dose group (Example 3), low-dose group (Comparative Example 1), low-dose group (Comparative Example 2), and low-dose group (Comparative Example 3). Except for the control group, all other groups received an intraperitoneal injection of 80 mg / kg for 3 days prior to the start of the experiment. -1 An immunosuppressive model was prepared using cyclophosphamide (CTX) once a day for 3 consecutive days, with a booster injection of the same dose on the 10th day of the experiment.

[0041] 3. Administration method: Nebulized administration began on the 4th day. Example 1 low-dose group: Nebulized administration of the herbal nebulizer solution from Example 1; 0.5 g / kg based on raw herbs. -1 Administer once daily for 30 consecutive days; In Example 1, the medium-dose group received the herbal nebulizer solution from Example 1 via nebulization; based on crude drug, 1 g·kg -1 Administer once daily for 30 consecutive days; Example 1 High-dose group: Nebulized administration of the traditional Chinese medicine nebulizer solution from Example 1; based on raw drug, 3 g·kg -1 Administer once daily for 30 consecutive days; Example 2 Low-dose group: Nebulized administration of the herbal nebulizer solution from Example 2; 0.5 g / kg based on raw herbs. -1 Administer once daily for 30 consecutive days; Example 3 Low-dose group: Nebulized administration of the traditional Chinese medicine nebulizer solution of Example 3; 0.5 g / kg based on raw herbs. -1 Administer once daily for 30 consecutive days; Comparative Example 1, low-dose group: administered the traditional Chinese medicine nebulizer solution of Comparative Example 1 via nebulization; based on crude drug, 0.5 g / kg -1 The drug was administered once daily for 30 consecutive days; Comparative Example 2 (low-dose group): the herbal nebulizer solution of Comparative Example 2 was administered via nebulization; based on crude drug, 0.5 g / kg -1 Administer once daily for 30 consecutive days; Comparative Example 3 low-dose group: Administered via nebulizer using the traditional Chinese medicine nebulizer solution from Comparative Example 3; 0.5 g / kg based on raw drug. -1 The drug was administered once daily for 30 consecutive days; the control and model groups were given the same amount of purified water; during the experiment, all mice were allowed free access to food and water.

[0042] 4. Mouse organ index detection: After the last administration, the mice were fasted. After weighing, the mice were sacrificed, and the spleen and thymus were separated and weighed. The spleen index and thymus index were calculated separately using the following formulas: Spleen index = Spleen mass (mg) / Mouse body weight (g); Thymus index = Thymus mass (mg) / Mouse body weight (g). The experimental results are shown in Table 1.

[0043] Table 1 The spleen and thymus are core immune organs of the body, and their organ indices can effectively reflect the strength of an animal's immune function. As shown in Table 1, compared with the control group, the model group mice showed a significant decrease in body weight gain and spleen index, indicating the successful construction of the immunosuppression model. The spleen index of mice in each treatment group was significantly higher than that of the model group, indicating that the tested drugs had a certain restorative effect on immune function. Comparative Examples 1-2 showed a significant decrease compared to the low-dose group in Example 1, indicating that *Hypericum perforatum* fruit, as the principal drug, can invigorate blood and regulate qi, while *Hypericum esculentum* fruit, as the assistant drug, can regulate qi and relieve pain to harmonize the middle jiao qi mechanism; both play an important role in improving mouse immunity. Comparative Example 3 also showed a significant decrease compared to Example 1, indicating that the present invention uses *Polygonatum sibiricum*, deer antler, and *Hypericum perforatum* fruit as the principal drugs, and *Lycium barbarum*, maca, *Atractylodes macrocephala*, and *Hypericum esculentum* fruit as the assistant drugs; the combination of these Chinese herbs can enhance the immune function of mice.

[0044] 5. ConA-induced spleen lymphocyte proliferation experiment: After the last administration, mouse spleens were harvested under aseptic conditions and placed in a petri dish containing an appropriate amount of sterile Hank's solution. The cells were ground with sterile forceps to prepare a single-cell suspension, which was then filtered through a 200-mesh cell sieve to remove tissue residue. The cells were washed twice with Hank's solution and centrifuged at 1000 r / min for 10 min. The cells were collected, resuspended in complete culture medium, counted, and the cell concentration was adjusted to 3 × 10⁻⁶ cells / mL. 6 Cells / mL. The adjusted cell suspension was seeded into 96-well cell culture plates, 1 mL per well. Two wells were used for each sample: one well containing 75 μL ConA solution (final concentration 100 μg / mL) as a stimulation well, and the other well without stimulation as a self-control. The culture plates were incubated at 37℃ and 5% CO2 for 72 h. Four hours before the end of culture, 0.7 mL of supernatant was carefully aspirated from each well, and an equal volume (0.7 mL) of fresh RPMI 1640 complete culture medium was added. Cell viability was then measured using a CCK-8 assay kit. The optical density (OD) value of each well was read at 570 nm using a microplate reader, and the difference between the OD values ​​of the stimulation and control wells was used as the evaluation index of spleen lymphocyte proliferation capacity.

[0045] Lymphocyte proliferation rate = (OD value of ConA well - OD value of control well) / OD value of control well × 100%. The experimental results are shown in Table 2.

[0046] Table 2 Cellular immunity is an important component of the body's specific immune response, and the proliferation capacity of T lymphocytes is widely recognized as an important objective indicator for evaluating the level of cellular immunity. As shown in Table 2, the lymphocyte proliferation rate in the model group was significantly lower than that in the control group, indicating the successful construction of the immunosuppression model. Compared with the model group, the lymphocyte proliferation rate in mice treated with the herbal nebulizer solution of this invention was significantly increased. Comparative Examples 1-3 all showed varying degrees of reduction compared to the low-dose group in Example 1, indicating that the herbal nebulizer solution of this invention can enhance the ConA-induced proliferation capacity of splenic lymphocytes in mice.

[0047] 6. Assay of NK cell killing activity Target cell preparation: Before the experiment, YAC-1 target cells were passaged and expanded. After collecting the cells, they were washed three times with Hank's solution, resuspended in RPMI 1640 complete culture medium, and the cell concentration was adjusted to 4 × 10⁻⁶. 5 Cells / mL (i.e., target cell suspension), for later use.

[0048] After the last administration, mouse spleens were aseptically harvested and placed in a petri dish containing an appropriate amount of sterile Hank's solution. The spleens were then shredded with sterile forceps to prepare a single-cell suspension, which was filtered through a 200-mesh cell sieve to remove tissue fragments. After cell counting, the cell concentration was adjusted to 2 × 10⁻⁶ cells / mL with complete culture medium. 7 cells / mL (i.e., effector cell suspension).

[0049] NK cell activity assay: The experiment included three control wells: (1) Target cell natural release wells: 100 μL each of target cell suspension and complete culture medium were added; (2) Target cell maximum release wells: 100 μL each of target cell suspension and 1% NP-40 lysis buffer were added; (3) Reaction wells: 100 μL each of target cell suspension and effector cell suspension were added. The cells were then incubated at 37℃ and 5% CO2 for 4 h. After incubation, the culture plate was centrifuged at 1500 r / min for 5 min, and 100 μL of supernatant from each well was carefully aspirated and transferred to the reaction plate of the lactate dehydrogenase (LDH) release kit. The optical density (OD) value of each well was measured at 490 nm wavelength using a microplate reader according to the kit instructions.

[0050] NK cell killing activity was calculated using the following formula: NK cell activity (%) = (OD value of reaction well - OD value of spontaneous release well) / (OD value of maximum release well - OD value of spontaneous release well) × 100%. The experimental results are shown in Table 3.

[0051] Table 3 NK cells, as key effector cells in the body's non-specific immune defense system, play a crucial role in clearing foreign pathogens, recognizing and killing mutated and tumor cells, and maintaining the body's immune surveillance function. As shown in Table 3, the NK cell activity of the model group mice was significantly reduced compared to the control group. Compared to the model group, the NK cell activity of the mice treated with the herbal nebulizer solution of this invention was significantly increased. Specifically, comparative groups 1-3 all showed varying degrees of reduction in NK cell activity compared to the low-dose group in Example 1. This indicates that the group in Example 1 of this invention has a better effect on improving the cellular immunity of immunocompromised mice.

[0052] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. Based on the present invention, some modifications or substitutions can be made, but these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of protection claimed by the present invention.

Claims

1. A traditional Chinese medicine nebulizer solution for enhancing immunity, characterized in that, The ingredients include the following parts by weight: Polygonatum sibiricum 23-28 parts, deer antler 15-18 parts, Cynanchum paniculatum fruit 10-14 parts, wolfberry 9-13 parts, maca 11-14 parts, Atractylodes macrocephala 7-11 parts, tuna 3-5 parts, Ziziphus jujuba seed 5-8 parts, Ophiopogon japonicus 9-11 parts, Poria cocos 6-10 parts, and licorice 1-3 parts.

2. The traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 1, characterized in that, The herbal atomizing liquid comprises the following raw materials in parts by weight: 26 parts Polygonatum sibiricum, 17 parts deer antler, 11 parts of Cymbidium faberi fruit, 10 parts wolfberry, 13 parts maca, 8 parts Atractylodes macrocephala, 4 parts of Evodia rutaecarpa fruit, 6 parts Ziziphus jujuba seed, 10 parts Ophiopogon japonicus, 7 parts Poria cocos, and 3 parts licorice.

3. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 1 or 2, characterized in that, Includes the following steps: (1) Take each raw material according to the weight, remove impurities, clean, crush, add water to soak, decoct twice, collect the decoction, filter, and combine the decoctions; (2) Concentrate the medicinal liquid obtained in step (1) to obtain a concentrated liquid, then add ethanol to the concentrated liquid and let it stand for 24 hours, filter, concentrate the filtrate to obtain an extract, then add water to the extract to obtain a traditional Chinese medicine solution, mix evenly, filter, sterilize, and dispense to obtain a traditional Chinese medicine atomizing liquid.

4. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 3, characterized in that, Step (1) The particle size of each raw material after pulverization is 50-80 mesh.

5. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 3, characterized in that, The amount of water added is 10-13 times the total mass of all raw materials.

6. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 3, characterized in that, The soaking time in step (1) is 20-30 minutes.

7. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 3, characterized in that, The simmering temperature in step (1) is 90-100℃ and the time is 25-30 minutes.

8. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 3, characterized in that, The relative density of the concentrate in step (2) is 1.0-1.2; the volume of ethanol accounts for 50-60% of the volume of the concentrate; and the relative density of the extract is 1.1-1.

3.

9. The method for preparing a traditional Chinese medicine nebulizer solution for enhancing immunity according to claim 3, characterized in that, In step (2), the ratio of extract to water in the herbal solution is (3-5) g: 100 mL.

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