Dendrobium officinale tender shoots rich in active ingredients and application of dendrobium officinale tender shoots

Abstract

The invention discloses dendrobium officinale tender shoots with enriched active ingredients, and belongs to the technical field of functional health care products. The dendrobium officinale tender shoot product is prepared by combining dendrobium officinale tissue culture with a dynamic gas regulation and control liquid suspension culture technology, the culture process comprises three stages of protocorm induction, tender shoot differentiation and proliferation and amplification, and the specific accumulation of 2-amino-1, 3, 5, 6-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1 The preparation method comprises the following steps: preparing characteristic metabolites such as 1, 2, 4-octadecane triol, 5-hydroxymethylfurfural, malic acid, adenosine, erucyl amide and 4-oxoproline; compared with a traditional mature plant, the dendrobium officinale tender shoot secondary metabolite pedigree is remarkably different, and the core active component of the dendrobium officinale tender shoot secondary metabolite pedigree shows breakthrough pharmacological activity in the directions of anti-inflammation, chemotherapy synergistic interaction, toxicity relief and the like; based on the production advantage that the cost of the raw materials is greatly reduced, the technology provides an industrialization path for developing novel medicinal and edible products instead of mature dendrobium officinale, and efficient and general application of dendrobium officinale resources can be promoted.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional health products, and particularly relates to tender buds of Dendrobium officinale with enriched active ingredients prepared by a new method, and discloses the application of the tender bud product of Dendrobium officinale in medical aspects such as anti-inflammation, chemotherapy potentiation, and chemotherapy attenuation. Background Art

[0002] Inflammation is a protective mechanism for the body to resist the invasion of foreign pathogens and is an immune response generated after the body is stimulated by an antigen. And the phenomenon of fever is a secondary effect of inflammation. Currently, drugs clinically used for treating inflammation are usually non-steroidal anti-inflammatory drugs (NSAIDs), which can reduce the inflammatory response by inhibiting the cyclooxygenase (COX) pathway and non-prostaglandin-mediated effects. COX-2 is a subtype of COX, which is rarely expressed in normal cells and will increase in expression when cells are stimulated by inflammatory signals and the like. Highly expressed COX-2 can cause fever and pain by catalyzing the generation of prostaglandin substances such as PGE2 and PGF2. In addition, COX-2 can also cause the proliferation of tumor cells, and its high expression can lead to an increase in PGE2, thus causing cancerous fever. Therefore, COX-2 can be used as a target for treating inflammation and tumors. However, NSAIDs are prone to cause gastrointestinal bleeding, myocardial infarction, stroke, etc., and have side effects on the central nervous system.

[0003] In addition, chemotherapeutic drugs clinically used such as cisplatin and cyclophosphamide (CTX) exhibit nephrotoxicity and myelosuppressive effects. The use of CTX will cause a decrease in the number of immune cells such as white blood cells, neutrophils, and lymphocytes, thereby inhibiting the body's immune function. At the same time, CTX will also bring impacts such as weight loss, decreased food intake, and metabolic disorders of the body to the body. These are all problems that need to be urgently solved in the clinical use of chemotherapy drugs such as CTX.

[0004] Currently, most studies focus on finding anti-inflammatory and post-chemotherapy recovery drugs with better efficacy and safety from natural products. Relevant studies have pointed out that Dendrobium officinale has various pharmacological effects such as anti-inflammatory, antioxidant, regulating glycolipid metabolism, protecting the nervous system, and anti-tumor. The active ingredients that play these pharmacological roles mainly include alkaloids, polysaccharides, bibenzyls, phenanthrenes, and fluorenones. For example, Zheng Zhewen et al. pointed out in the article "Research Progress and Clinical Application of Dendrobium officinale in Inhibiting Breast Cancer" that Dendrobium officinale Fengdoujing can enhance the anti-tumor effect of chemotherapy drugs, has the effects of enhancing efficacy and detoxifying, can improve the survival rate, extend the survival period, and also has a certain protection against radiation damage. In addition, the English literature "ZENG Q, et al. Inhibitory effect of different Dendrobium species on LPS-induced inflammation in macrophages via suppression of MAPK pathways[J]. Chinese journal of natural medicines, 2018, 16(7): 481-489" pointed out that different Dendrobium species can inhibit the inflammation of lipopolysaccharide (LPS)-induced macrophages, can inhibit the expression of COX-2 and the phosphorylation of ERK and JNK, and Dendrobium officinale has the best effect due to its rich polysaccharides and phenols.

[0005] Although the existing technology shows that Dendrobium officinale has good medicinal values such as anti-inflammatory and anti-tumor, its industrial application still faces severe bottlenecks: 1) Low efficiency of raw material acquisition: The seeds of Dendrobium are extremely small, without endosperm, and have harsh growth conditions. Even through artificial tissue culture technology (which needs to go through 6-8 stages such as protocorm induction, proliferation and differentiation, and acclimatization transplantation), it still takes a long cycle from seeds to mature plants; 2) High cost conduction pressure: The medicinal components are concentrated in the mature stems (which need to be processed into Fengdoujing or extracts). Coupled with the low-yield characteristics, the raw material cost is as high as 300-500 yuan per kilogram, and the premium of the final product can reach more than 10 times, directly pushing up the economic burden on patients.

[0006] Relevant studies have shown that the chemical components and pharmacological activities of the tender buds of Dendrobium officinale are similar to those of mature plants, which provides a feasible basis for using the tender bud part to replace the mature stem as a medicinal raw material; at the same time, the tender bud propagation technology based on the in vitro cell culture system can shorten the production cycle to 3-4 months, and the cost of the active ingredients per unit yield decreases significantly, greatly alleviating the pressure on raw material supply and laying a technical foundation for the large-scale development of high-cost-effective functional products.

[0007] However, one point that needs attention is that although there are certain similarities in some chemical components between the tender buds and mature plants of Dendrobium officinale, the significant differences in their core components are still the key technical bottlenecks restricting the substitution of tender buds of Dendrobium officinale for mature Dendrobium. The main medicinal value of mature Dendrobium officinale stems from its high content of polysaccharides (22%-49% dry weight) and alkaloids (such as dendrobine and dendroside). Among them, polysaccharides can play immunomodulatory and anti-tumor roles by activating macrophages and T lymphocytes, while the synergistic effect of alkaloids and flavonoids can significantly improve oxidative stress and insulin resistance. In contrast, the polysaccharide content in the tender buds of Dendrobium officinale obtained by existing technologies is only 3%-8%, and the total alkaloid content is less than 20% of that of mature plants. Its component characteristics are more inclined to the accumulation of low-molecular-weight metabolites - the total amino acid content increases by 30%-50% (such as glutamic acid and aspartic acid), and the colloid content decreases by more than 70%. This makes the tender buds of Dendrobium officinale more suitable for gastric mucosa repair rather than high-intensity pharmacological interventions such as anti-tumor. Therefore, under the current technical system, mature Dendrobium is preferentially used for the treatment of severe diseases (such as tumor adjuvant therapy and diabetes complication conditioning) due to the high abundance of active ingredients, while the tender buds of Dendrobium officinale are positioned for the development of daily stomach-nourishing functional foods due to their mild characteristics.

[0008] To effectively improve the content and diversity of functional active ingredients in the tender buds of Dendrobium officinale, and make its key medicinal effects such as anti-inflammatory, synergistic enhancement of chemotherapy and toxicity alleviation reach or approach the level of mature Dendrobium, it is urgent to break through the secondary metabolism bottleneck of tender buds by optimizing cultivation techniques, so as to achieve the substitution of medicinal value while reducing production costs and shortening the growth cycle, and promote the efficient and inclusive application of Dendrobium officinale resources. Summary of the Invention

[0009] Aiming at the above existing problems, the present invention aims to provide a tender bud of Dendrobium officinale with enriched active ingredients and its application. Cultivating the tender buds of Dendrobium officinale by the method disclosed in the present application can not only shorten the cultivation cycle and reduce the cultivation cost, but also enrich specific secondary metabolites in a targeted manner. The obtained tender bud products of Dendrobium officinale show significant performance in core pharmacological dimensions such as anti-inflammatory, synergistic enhancement of chemotherapy and toxicity alleviation, providing an industrialization basis for developing high-cost-effective medicinal products and functional foods to replace traditional medicinal parts, and can promote the inclusive application of Dendrobium officinale resources.

[0010] To achieve the above purpose, the technical solution adopted by the present invention is as follows: A tender bud of Dendrobium officinale with enriched active ingredients, which is prepared by using the tissue culture of Dendrobium officinale combined with the liquid suspension culture technology of dynamic gas regulation. The cultivation process goes through three stages: protocorm induction, tender bud differentiation, and proliferation and amplification. The characteristic metabolites in the tender buds of Dendrobium officinale include 2-amino-1,3,4-octadecatriol, 5-hydroxymethylfurfural, malic acid, adenosine, erucamide, 4-oxoproline, 9-hydroxy-10,12-octadecadienoic acid, and citric acid.

[0011] Furthermore, during the preparation process, a weak sugar liquid medium is used in the tender bud induction and proliferation stages, and air and carbon dioxide are continuously introduced during the cultivation in these two stages.

[0012] The above-mentioned Dendrobium officinale tender buds enriched with active ingredients can be applied in the preparation of anti-inflammatory and antipyretic preparations, and the dosage form of the preparation is any one of powder, tablet, paste, capsule, granule, syrup, pill or oral film.

[0013] The above-mentioned Dendrobium officinale tender buds enriched with active ingredients can be applied in the preparation of chemotherapy adjuvant preparations, and the preparation has the dual functions of enhancing the efficacy of chemotherapy drugs and reducing their toxic and side effects. The drug delivery forms of the preparation include but are not limited to oral solid preparations, mucosal administration preparations or immediate-release / sustained-release composite dosage forms.

[0014] This application also discloses an inflammation-regulating functional health product, which contains a therapeutically effective amount of the above-mentioned Dendrobium officinale tender buds enriched with active ingredients as the core active component, and food-acceptable excipients.

[0015] This application also discloses a chemotherapy synergistic adjuvant preparation, which contains the above-mentioned Dendrobium officinale tender buds enriched with active ingredients, and the components of which achieve the directional enrichment of specific secondary metabolites through a dynamic gas regulation culture process.

[0016] Furthermore, the chemotherapy synergistic adjuvant preparation is combined with a chemotherapy drug to form a therapeutic composition, and the chemotherapy drug is selected from at least one of platinum compounds, alkylating agents and antimetabolic drugs.

[0017] Furthermore, after drying and grinding the above-mentioned Dendrobium officinale tender buds enriched with active ingredients, and then mixing some food-acceptable excipients, it can be used as a functional solid beverage; or directly juicing the fresh above-mentioned Dendrobium officinale tender buds enriched with active ingredients, and then mixing some food-acceptable excipients, it can be used as a functional liquid beverage; of course, when preparing solid or liquid beverages, excipients may not be added additionally, and only the Dendrobium officinale tender bud product is used as the only functional ingredient for direct consumption. Specifically, it depends on relevant functional requirements. Direct drinking by users or taking it after mixing with water can achieve beneficial effects such as inflammation regulation, chemotherapy synergistic enhancement and toxicity alleviation.

[0018] The beneficial effects of the present invention are:

[0019] 1. By optimizing the formula of the liquid medium for bud induction and proliferation, this application has successfully achieved the directional enrichment of active ingredients while significantly improving the proliferation efficiency of Dendrobium officinale buds and maintaining trait stability. As detected by LC-MS, the contents of medicinal ingredients such as 2-amino-1,3,4-octadecanetriol, 5-hydroxymethylfurfural, malic acid, adenosine, erucamide, and 4-oxoproline in the Dendrobium officinale buds cultivated in the improved medium have been significantly increased, laying a material foundation for the development of anti-inflammatory and antipyretic preparations and chemotherapy adjuvant drugs.

[0020] 2. Verified by pharmacodynamic experiments, the Dendrobium officinale buds cultivated by the method of this application can regulate the activity of COX-2 in a dose-dependent manner, thus exerting a definite anti-inflammatory and antipyretic effect.

[0021] 3. The Dendrobium officinale buds cultivated by the method disclosed in this application can improve the leukopenia symptoms of C57BL / 6 mice bearing LLC murine lung cancer caused by cyclophosphamide, and can also significantly reduce the volume of transplanted tumors in chemotherapy-bearing tumor mice. It can also effectively improve the problem of reduced appetite in BALB / c nude mice bearing A549 human lung cancer caused by cyclophosphamide, reduce the volume of transplanted tumors in chemotherapy-bearing tumor nude mice, and prolong the survival period of chemotherapy-bearing tumor nude mice. It shows that this Dendrobium officinale bud product has an enhancing effect on the antitumor effect of chemotherapy drugs, has the effects of enhancing efficacy and detoxifying, can improve the survival rate of patients, prolong the survival period, and has very good medical prospects.

[0022] 4. For the Dendrobium officinale buds cultivated by the method disclosed in this application, there are significant differences in the core functional active ingredients and the secondary metabolite profiles of mature plants, but they show excellent performance in pharmacological dimensions such as anti-inflammatory, synergistic enhancement of chemotherapy efficacy, and toxicity alleviation. Research has confirmed that using these Dendrobium officinale buds to replace traditional mature plants to develop medicinal and health products not only has feasibility advantages and broad application prospects, but also can promote the inclusive application of Dendrobium officinale resources by reducing raw material costs.

[0023] 5. This application optimizes the tissue culture system of Dendrobium officinale buds by using an improved MS medium combined with dynamic gas regulation (continuously introducing air and carbon dioxide), synergistically shortening the proliferation cycle and the overall cultivation time during the bud induction and proliferation stages, significantly improving the large-scale production efficiency, reducing the production cost, and at the same time enhancing the market supply response ability, providing reliable technical support for the efficient market application of Dendrobium officinale bud resources. Description of the Drawings

[0024] Figure 1 A photo of the Dendrobium officinale buds cultivated in Example 1;

[0025] Figure 2 It is a schematic structural diagram of a cultivation barrel, in which 1 is a cultivation barrel, 2 is an air inlet pipe, 3 is a carbon dioxide inlet pipe, 4 is an exhaust pipe, and 5 is a feed pipe.

[0026] Figure 3 is Figure 2 The large picture of part A in it;

[0027] Figure 4 It is a photo of the tender buds of Dendrobium officinale after drying;

[0028] Figure 5 It is the base peak chromatogram in the positive ion mode of the sample;

[0029] Figure 6 It is the base peak chromatogram in the negative ion mode of the sample;

[0030] Figure 7 It is the COX-2 concentration in the hypothalamus of mice in different groups, where indicates the significance of difference P < 0.001 compared with the blank control group, *** indicates the significance of difference P < 0.001 compared with the model group, and +++ indicates the significance of difference P < 0.001 compared with the low-dose tender bud group;

[0031] Figure 8 It is the statistical chart of the tumor volume of mice in each group, and *** indicates the significance of difference P < 0.001 compared with the model group;

[0032] Figure 9 It is the food intake and survival curve of nude mice in each group. Among them, subfigure A is the average diet weight of nude mice in each group during the recovery period (7 days) after each round of chemotherapy, subfigure B is the survival curve of nude mice in each group, * indicates the significance of difference P < 0.05 compared with the model group, ** indicates the significance of difference P < 0.01 compared with the model group, and the red arrow represents the time point of each round of chemotherapy;

[0033] Figure 10 It is the tumor volume of mice in different groups after different rounds of chemotherapy. * indicates the significance of difference P < 0.05 compared with the model group, and *** indicates the significance of difference P < 0.001 compared with the model group. Detailed implementation method

[0034] In order to enable ordinary technicians in the field to better understand the technical solution of the present invention, the technical solution of the present invention will be further described below in conjunction with the drawings and embodiments.

[0035] Embodiment 1

[0036] This embodiment discloses a kind of tender buds of Dendrobium officinale with enriched active ingredients. The cultivation process of the tender buds of Dendrobium officinale specifically includes several culture stages such as protocorm induction - tender bud induction - tender bud proliferation, and different culture stages will apply protocorm induction medium, tender bud induction liquid medium and tender bud proliferation liquid medium.

[0037] Among them, the following elemental stock solutions are required for preparing the tender shoot induction liquid medium and the tender shoot proliferation liquid medium:

[0038] The first organic elemental stock solution ( / L): glycine 1 g, pyridoxine hydrochloride 0.25 g, thiamine hydrochloride 0.05 g, nicotinic acid 0.25 g, inositol 50 g, and the balance is deionized water;

[0039] The sulfur, nitrogen, and phosphorus elemental stock solution ( / L): magnesium sulfate (MgSO4·7H2O) 37 g, potassium nitrate 190 g, potassium dihydrogen phosphate 17 g, and the balance is deionized water;

[0040] The calcium salt stock solution ( / L): calcium chloride (CaCl2·2H2O) 44 g, calcium nitrate 28 g, and the balance is deionized water;

[0041] The ammonium salt stock solution ( / L): ammonium sulfate 48.5 g, and the balance is deionized water;

[0042] The first iron salt stock solution ( / L): ferrous sulfate (FeSO4·7H2O) 3.45 g, disodium ethylenediaminetetraacetate (Na2-EDTA) 3.73 g, and the balance is deionized water;

[0043] The trace element stock solution ( / L): manganese sulfate (MnSO4·4H2O) 16.9 g, zinc sulfate (ZnSO4·7H2O) 8.6 g, boric acid 6.2 g, potassium iodide 0.83 g, sodium molybdate 0.25 g, copper sulfate 0.025 g, cobalt chloride 0.025 g, and the balance is deionized water;

[0044] 6-Benzylaminopurine (6-BA) stock solution: 100 mg of 6-BA is dissolved in 10 mL of HCl solution (1 mol / L), and the volume is made up to 100 mL with deionized water;

[0045] Naphthaleneacetic acid (NAA) stock solution: 50 mg of naphthaleneacetic acid is dissolved in 20 mL of ethanol solution (95%), and the volume is made up to 100 mL with deionized water;

[0046] Potato extract: Wash and peel the potatoes and cut them into slices. Put 200 g of sliced potatoes into 400 mL of distilled water and cook for 25 min, then filter and collect the filtrate.

[0047] The formula of the tender shoot induction liquid medium is as follows:

[0048] Measure 350 mL of potato extract, add distilled water to make up to 600 mL, and sequentially add 35 mL of coconut water, 2 mL of the first organic element stock solution, 10 mL of the sulfur-nitrogen-phosphorus element stock solution, 10 mL of the calcium salt stock solution, 10 mL of the ammonium salt stock solution, 1 mL of the trace element stock solution, 10 mL of the first iron salt stock solution, 1 mL of the 6-BA stock solution, and 0.4 mL of the NAA stock solution. Add 18 g of sucrose, stir well to mix evenly, make up the volume to 1000 mL with distilled water, adjust the pH of the solution to 6.10, and sterilize at 121 °C and 1.1 MPa for 21 minutes to obtain the shoot induction liquid medium. After sterilization, the pH of the medium will drop to 5.6 - 5.8.

[0049] The formula of the shoot proliferation liquid medium is as follows:

[0050] Measure 350 mL of potato extract, add distilled water to make up to 600 mL, and sequentially add 35 mL of coconut water, 2 mL of the first organic element stock solution, 10 mL of the sulfur-nitrogen-phosphorus element stock solution, 10 mL of the calcium salt stock solution, 10 mL of the ammonium salt stock solution, 1 mL of the trace element stock solution, 10 mL of the first iron salt stock solution, 0.3 mL of the 6-BA stock solution, 7 g of sucrose, and 5 g of edible glucose. Stir well to mix evenly, make up the volume to 1000 mL with distilled water, adjust the pH of the solution to 6.10, and sterilize at 121 °C and 1.1 MPa for 21 minutes to obtain the shoot proliferation liquid medium.

[0051] The following stock solutions are required for preparing the protocorm induction medium:

[0052] The second organic element stock solution ( / L): glycine 0.2 g, pyridoxine hydrochloride 0.05 g, thiamine hydrochloride 0.01 g, nicotinic acid 0.05 g, inositol 10 g, the balance is deionized water;

[0053] The macroelement stock solution ( / L): potassium nitrate 38 g, ammonium sulfate 16.5 g, magnesium sulfate (MgSO4·7H2O) 7.4 g, potassium dihydrogen phosphate 3.4 g, calcium chloride (CaCl2·2H2O) 8.8 g, the balance is deionized water;

[0054] The trace element stock solution: the same as the trace element stock solution used in preparing the shoot induction liquid medium;

[0055] The second iron salt stock solution ( / L): ferrous sulfate (FeSO4·7H2O) 5.56 g, disodium ethylenediaminetetraacetate (Na2-EDTA) 7.46 g, the balance is deionized water;

[0056] The naphthylacetic acid (NAA) stock solution: 50 mg of naphthylacetic acid is dissolved in 20 mL of ethanol solution (95%), and the volume is made up to 100 mL with deionized water.

[0057] The formula of the protocorm induction medium is as follows:

[0058] Add 75 g of water to 30 g of potatoes after cleaning and peeling, and process them with a juicer to obtain mashed potatoes. Pour the mashed potatoes into a preparation container, and sequentially add 25 mL of macroelement mother liquor, 1 mL of microelement mother liquor, 10 mL of second organic element mother liquor, 5 mL of second iron salt mother liquor, 0.2 mL of naphthaleneacetic acid mother liquor, 30 g of sucrose, and 5 g of agar powder. Stir and mix well, make up the volume to 1000 mL with distilled water, adjust the pH to 5.9, and sterilize at 121 °C for 21 min to obtain the protocorm induction medium.

[0059] The specific process of cultivating Dendrobium officinale buds is as follows:

[0060] S1. Select mature and unopened Dendrobium officinale seeds, inoculate them into the protocorm induction medium after disinfection treatment, and culture them at a temperature of 20-26 °C, with a relative air humidity of 55-65%, a light time of 10 h / day, and a light intensity maintained at 1500 Lux to obtain Dendrobium officinale protocorms; it is found through testing that the temperature and humidity conditions will fluctuate to a certain extent with the adjustment of the light period and dark period, but as long as the fluctuations are within the above range, the impact on the final culture effect is not significant.

[0061] S2. Inoculate the Dendrobium officinale protocorms obtained in the previous step into the bud induction liquid medium at an inoculation density of 12 g / L. The medium is placed in a relatively sealed culture barrel 1 (the structure of the culture barrel refers to Figure 2 、 Figure 3)It enables the young shoot induction process to be completed in a relatively enclosed environment to reduce the influence of the external environment on the culture process. The filling volume of the culture medium in the barrel is 8.5 L. When the amount of the culture medium is less than 6 L, liquid supplement is required, and the liquid supplement volume is 4 L (determined by the liquid level scale on the barrel body). The culture temperature is 21 - 26 °C, the relative air humidity is 45 - 55%, the light time is 12 h / day, and the light intensity is 2600 Lux (the temperature and humidity fluctuations within the corresponding range have little impact. In principle, the temperature is slightly higher during the light period and slightly lower during the dark period); An air inlet pipe 2, a carbon dioxide inlet pipe 3, an exhaust pipe 4, and a feed pipe 5 that penetrate inside and outside the barrel body are connected to the culture barrel 1. One-way valves are provided on different pipes. The air inlet pipe 2 and the carbon dioxide inlet pipe 3 are inserted below the liquid level. The feed pipe 5 is used to input protocorms into the barrel. After the explants are introduced, the one-way valve on the feed pipe 5 is closed; During the young shoot induction culture process of Dendrobium officinale, filtered compressed air is injected into the liquid culture medium in the culture barrel 1 through the air inlet pipe 2, the ventilation volume is 3 L / min, and the flow rate is 12 m / s; While injecting air, carbon dioxide gas is additionally injected into the culture barrel 1 through the carbon dioxide inlet pipe 3, and the ventilation flow rate is 11 L / h. A pressure detection device is provided inside the barrel. When there are too many harmful gases inside the barrel, the pressure inside the barrel will increase. After reaching the set threshold, the one-way valve on the exhaust pipe 4 is opened, and the exhaust gas inside the barrel is discharged through the exhaust pipe 4 by using a power device such as an air pump. When the pressure inside the barrel does not reach the set threshold range, the one-way valve on the exhaust pipe 4 is closed. The corresponding valves on the culture barrel 1 can be manually adjusted or uniformly controlled by the main controller for automatic regulation. During the culture process, the feed pipe 5 is closed, the exhaust pipe 4 is not connected to the outside environment in a non-exhaust state, and the two inlet pipes 5 are in the air intake (filtered air and carbon dioxide) state and are not directly connected to the outside environment. Therefore, the inside of the culture barrel 1 is in a relatively sealed state to prevent external impurities from falling into the barrel body and polluting the culture environment.

[0062] Preferably, a gas distribution device is provided at the air outlet ends of the air inlet pipe 2 and the carbon dioxide inlet pipe 3 to disperse the gas input into the barrel, improve the uniformity of gas distribution, and thus help to improve the gas utilization rate and the cultivation effect.

[0063] During the culture process, continuously injecting air into the culture barrel 1 can mainly keep the protocorms suspended in the liquid culture medium in a state of turning, prevent the protocorms from agglomerating in the culture medium and causing their growth to be inhibited, maintain the uniformity of the suspension culture, and improve the nutrient absorption rate; and continuously introducing fresh air during the culture process can take away some waste gas produced by plant metabolism, which can prevent the local gas concentration imbalance from affecting cell growth and ensure the cleanliness of the culture environment; in addition, after the air is introduced, the dissolved oxygen concentration in the liquid culture medium can be increased, providing the necessary conditions for the aerobic respiration of the protocorms, thereby providing energy for the absorption of nutrients by Dendrobium officinale. So in general, continuously injecting fresh air during the culture process can maintain the respiration, gas exchange, metabolic balance of the protocorms and the dispersion state of the protocorms, thereby improving the nutrient absorption rate of the protocorms.

[0064] In addition, although the injected air contains a certain amount of carbon dioxide, it is not efficient to replenish carbon dioxide into a relatively closed barrel by simply filling it with air. Injecting carbon dioxide into the barrel while injecting air can make up for the problem of insufficient carbon dioxide, ensure an adequate supply of raw materials required for photosynthesis, promote the photosynthesis efficiency of the protocorm and optimize the metabolic environment. The increase in carbon dioxide concentration in the barrel can significantly increase the photosynthesis rate of the protocorm and promote the plant to synthesize more sugars and organic matter, thereby accelerating the growth rate of Dendrobium officinale shoots and enhancing biomass accumulation.

[0065] In addition, the additional addition of carbon dioxide during the tissue culture process can also replace part of the carbon source, breaking the limitations of the traditional culture method that mainly relies on sugar as a carbon source in terms of high cost, difficulty in further improving efficiency, and high pollution rate, and helping to successfully transform into a CO2 weak sugar mixotrophic model, that is, it is not to completely replace the carbon source with CO2 to establish a cultivation model that completely changes from heterotrophy to autotrophy, but to use CO2 to replace part of the carbon source for cultivation. A new weak sugar mixotrophic model that has both autotrophy and heterotrophy. The shortcomings of transitional reliance on sugar as a carbon source are high cost and easy to cause microbial contamination problems. The present application injects an appropriate amount of CO2 gas into the culture barrel 1 during the culture process to replace part of the carbon source, thereby optimizing the carbon source supply. Then, by accurately adjusting environmental factors such as light, the tissue culture cost can be controlled and the quality of the young shoots of Dendrobium officinale can be effectively improved.

[0066] Specifically, the concentration of carbon dioxide in the barrel needs to be appropriately adjusted according to the light-dark cycle. Since the protocorms of Dendrobium officinale may have insufficient carbon dioxide absorption due to low stomatal opening during the light period, and absorb and store carbon dioxide through open stomata during the dark period for photosynthesis during the light period, during the light period culture, the carbon dioxide concentration in the barrel can be appropriately increased to 600-800 ppm (the carbon dioxide concentration cannot reach an exact and fixed value due to changes in gas temperature and pressure, mainly serving as a reference for adjustment) to make up for the insufficient absorption efficiency during the light period; during the dark period, it can be reduced to 300-500 ppm, and the concentration is monitored by a sensor set in the barrel to maintain the basic metabolic requirements.

[0067] After about 30 days of culture, the induction process of Dendrobium officinale tender buds can be completed, and induced cormels can be obtained in this step.

[0068] S3. Transfer the cormels obtained in S2 to a liquid medium for tender bud proliferation for proliferation culture. The inoculation density is 10 g / L, and the medium is also placed in the culture barrel 1 described in step S2. The filling volume of the medium in the barrel is 8 L. When the amount of the medium is less than 6 L, replenishment is required, and the replenishment volume is 3 L. The culture temperature is 20-26 °C, the relative air humidity is 55-60%, the light time is 16 h / day, and the light intensity is 3000 Lux (temperature and humidity fluctuations within the corresponding ranges have little impact. In principle, the temperature is slightly higher during the light period and slightly lower during the dark period); an air inlet pipe 2, a carbon dioxide inlet pipe 3, an exhaust pipe 4, and a feed pipe 5 that penetrate inside and outside the barrel body are also connected to the barrel. The corresponding operations are the same as in step S2. The feed pipe 5 is used to inject cormels into the barrel. During the proliferation culture of Dendrobium officinale tender buds, filtered compressed air is injected into the culture barrel through the air inlet pipe 2, and the ventilation volume is 4 L / min, and the flow rate is 10 m / s; and carbon dioxide gas is synchronously injected into the culture barrel 1 through the carbon dioxide inlet pipe 3, and the ventilation flow rate is 13 L / h. The exhaust pipe 4 is used to discharge the waste gas in the barrel. After about 15 days of culture, Dendrobium officinale tender buds can be obtained. For a photo of the freshly cultivated Dendrobium officinale tender buds, refer to Figure 1 。

[0069] Detection of Active Ingredients in Dendrobium officinale Tender Buds

[0070] The freshly prepared Dendrobium officinale tender buds in Example 1 are dried. For a photo of the dried Dendrobium officinale tender buds, refer to Figure 4 , take 20 mg of the dried Dendrobium officinale tender buds, add 500 μL of methanol, ultrasonically extract for 30 minutes, centrifuge at 13000 rpm for 10 minutes, take the supernatant, and perform on-machine detection (LCMS).

[0071] The liquid chromatography instrument is: UltiMate3000 UHPLC, and the chromatographic column is Thremo Hypersil gold C18 (1.9μm, 2.1mm×100mm). The flow rate for analysis is 0.3 mL / min, and the injection volume is 10 μL. The mobile phase is 0.1% formic acid / acetonitrile (B) - 0.1% formic acid / water (A).

[0072] The mass spectrometry instrument is Q-Exactive (Thermo Fisher Scientific, CA, USA) with HESI source, ion source temperature 310°C, capillary temperature 320°C, sheath gas flow rate 30 units, auxiliary gas flow rate 10 units, spray voltage 3 kV in positive ion mode, and spray voltage 2.8 kV in negative ion mode.

[0073] Analysis uses data-dependent scanning analysis (DDA), the loop count set value is 10, the HCD energy uses step-wise normalized collision energy, the set values are 10, 28, 35 eV. The first-order mass spectrometry scan range is 100 - 1500 m / z, the resolution is set to 70000, the AGC target is set to 3E6, and the injection time is set to 200 ms. The second-order mass spectrometry resolution is set to 17500, the AGC target is set to 1E5, and the injection time is set to 50 ms.

[0074] The base peak chromatograms of the sample in positive and negative ion modes are shown in Figure 5 and Figure 6 . The LCMS data is automatically searched against a database (including chemspider plant database CHEBI, CHEMBL, natural product database, flavonoid database, OTC traditional Chinese medicine database, and mz cloud database, etc.) using Compounddiscover software (V3.2, Thermo Fisher Scientific, CA, USA) to analyze the possible chemical components. Chemical components with higher matching degrees or higher contents (peak area percentage greater than 1.5%) are retained. See Table 1 for details.

[0075] Table 1. Chemical Component Analysis

[0076]

[0077] According to the data in Table 1, the contents of 2-amino-1,3,4-octadecanetriol, 5-hydroxymethylfurfural, malic acid, adenosine, erucamide, 4-oxoproline, 9-hydroxy-10,12-octadecadienoic acid and citric acid in the sample are relatively high. According to relevant reports, these compounds show remarkable performance in the medical field. For example, 2-amino-1,3,4-octadecanetriol has the functions of moisturizing, antibacterial and promoting skin repair; 5-hydroxymethylfurfural plays a role in protecting the liver and reducing blood pressure by antioxidant and improving blood circulation; malic acid can promote digestion, assist in regulating acid-base balance, increase appetite and adjust taste; adenosine can relieve cardiovascular diseases; erucamide has antibacterial and antioxidant effects; 4-oxoproline, as an immunomodulator, affects the functions of T lymphocytes and macrophages and can enhance the body's anti-infection ability; 9-hydroxy-10,12-octadecadienoic acid (HODE) can reduce chronic inflammatory responses by inhibiting the release of inflammatory factors or regulating the functions of immune cells, and may have intervention value for autoimmune diseases or inflammatory bowel diseases. Moreover, HODE is used to analyze abnormal lipid metabolism pathways and assist in the analysis of disease mechanisms (such as metabolic syndrome, tumor metabolic reprogramming, etc.); the medicinal value of citric acid covers multiple fields such as digestion, urinary system, cardiovascular and immune regulation, and is widely used as a pharmaceutical excipient in the development of preparations.

[0078] It should be noted that the above components have not been widely reported in mature Dendrobium officinale plants and Dendrobium officinale buds cultivated by conventional cultivation techniques. Their significant enrichment in the Dendrobium officinale buds prepared in this application not only provides new ideas for medicinal directions such as anti-inflammatory, synergistic enhancement of chemotherapy and toxicity alleviation, but also indicates the potential feasibility of replacing mature plants with the newly cultivated Dendrobium officinale buds to develop medicinal and functional health products.

[0079] Application Example

[0080] Based on the above component analysis, it can be seen that the Dendrobium officinale buds cultivated according to the method disclosed in Example 1 contain a variety of beneficial functional components and show good application prospects in the medical field. Therefore, in this application example, the synergistic effect of Dendrobium officinale buds on chemotherapy in tumor-bearing animals and its toxicity reduction effect were further studied.

[0081] 1. Experimental Materials

[0082] 1.1 Experimental Animals

[0083] Provided by Liaoning Changsheng Biotechnology Co., Ltd., with the license number SCXK(Liao)2020-0001. SPF-grade KM mice, male, 4 weeks old, 18-22 g. SPF-grade C57BL / 6 mice, male, 4 weeks old, 18-22 g. SPF-grade BALB / c nude mice, male, 4 weeks old, 22-25 g.

[0084] 1.2 Test Articles

[0085] Test article: The dried tender buds of Dendrobium officinale were ground into powder and suspended with sodium carboxymethylcellulose (CMC-Na) to obtain the test article, and the content of the powder of Dendrobium officinale tender buds was ≥ 99.5%.

[0086] 2. Experimental methods

[0087] 2.1. Antipyretic experiment

[0088] After 48 KM mice adapted to the environment, they were randomly divided into four groups (each group was divided into two cages, with six mice in each cage): blank control group (0.5% CMC-Na), model group (0.5% CMC-Na), low-dose tender bud group (1170 mg / kg / d), and high-dose group (3510 mg / kg / d). The mice were intragastrically administered once a day at an equal volume of 0.2 mL / 10 g body weight for 14 consecutive days. Among them, on the 9th, 10th, and 11th days of drug administration, the mice (except the blank control group) were intraperitoneally injected with cyclophosphamide (CTX) at 100 mg / kg once a day for 3 consecutive days.

[0089] Determination of hypothalamic COX-2

[0090] 1) Sample collection: The mice were anesthetized by intraperitoneal injection of 6% chloral hydrate solution and then fixed. The abdominal midline and sternum were cut open to fully expose the heart. A needle was inserted into the apex of the heart, and a small incision was made in the right auricle. The heart perfusion was completed by rapidly perfusing 20 mL of sodium chloride injection until the liver was completely bloodless and turned white. Subsequently, the mouse brain was dissected, and the hypothalamus was removed and placed in an EP tube. After all the tissues were collected, they were stored at -80 °C for later use.

[0091] 2) ELISA detection: According to the instructions of the COX-2 detection kit, 100 μL of diluted sample or standard products with different concentrations was added to each well, and after covering with a sealing plate membrane, it was incubated at 37 °C for 1 h. After the incubation ended, the liquid was discarded. 100 μL of biotinylated antibody working solution was added to each well and incubated again at 37 °C for 1 h. Subsequently, the liquid was discarded. 300 μL of 1× washing solution was added to each well and allowed to stand for 1 min, and then the washing solution was discarded and patted dry on absorbent paper. After repeating the washing 3 times, 100 μL of enzyme conjugate working solution was added to each well and incubated at 37 °C for 30 min. The previous washing operation was repeated, and a total of 5 washes were performed. Subsequently, 90 μL of substrate TMB was added to each well, and it was incubated at 37 °C in the dark for 15 min. Finally, 50 μL of stop solution was added to each well, and the OD value of each well was immediately measured at a wavelength of 450 nm.

[0092] COX-2 is an inducible enzyme that is less expressed in normal tissues but highly expressed when cells are stimulated by inflammation. The increase in COX-2 expression easily promotes the release of inflammatory factors, which increase body temperature by affecting the temperature regulation center in the hypothalamus. CTX is a broad-spectrum anti-tumor drug and immunosuppressant. In animal experiments, CTX is often used to prepare a chemotherapy-induced myelosuppression model. Therefore, detecting the level of COX-2 in the hypothalamus of mice can be used to investigate the antipyretic effect of the tender buds of Dendrobium officinale on mice after chemotherapy. The results are shown in Figure 7 , compared with the blank control group, the level of COX-2 in the hypothalamus tissue of the model group mice increased significantly; compared with the model group, the concentrations of COX-2 in the hypothalamus tissues of the low- and high-dose tender bud groups of mice decreased significantly, and the intervention effect was obvious, showing a dose-dependent improvement, suggesting that the tender buds of Dendrobium officinale have a good antipyretic effect on mice after chemotherapy.

[0093] 2.2 Experimental study on leukopenia in mice bearing LLC murine lung cancer cells

[0094] After 60 C57BL / 6 mice adapted to the environment, they were randomly divided into five groups (each group was divided into two cages, with six mice in each cage): blank control group (0.5% CMC-Na), model group (0.5% CMC-Na), Sanguisorba officinalis group (156 mg / kg / d), low-dose tender bud group (1170 mg / kg / d), and high-dose tender bud group (3510 mg / kg / d). Subcutaneous injection of LLC murine lung cancer cells was used to inoculate the mice in the model group and each drug administration group to establish an ectopic transplantation tumor model. After 24 hours of tumor inoculation, drug administration was carried out. The mice were gavaged once a day at an equal volume of 0.2 mL / 10 g body weight for 14 consecutive days. Among them, on the 9th, 10th, and 11th days after drug administration, mice (except the blank control group) were intraperitoneally injected with CTX 100 mg / kg once a day for 3 consecutive days. Three days after the last drug administration, the tumor was measured with a vernier caliper.

[0095] The specific method for establishing an ectopic transplantation tumor model of LLC murine lung cancer cells is as follows: Take LLC murine lung cancer cells in the logarithmic growth phase, digest them with trypsin and then centrifuge to collect. After collection, resuspend the cells with PBS, count them, and adjust the cell number to 1×10 7 cells / mL. Inject 100 μL of the above cell suspension into the right axilla of each mouse to complete the construction of the transplantation tumor model of LLC lung cancer cell line.

[0096] Blood routine test: 2 hours after drug administration on the 14th day, 0.5 mL of orbital blood was collected from each group of mice and injected into a 1.5 mL EDTA anticoagulation tube. Shake the centrifuge tube up and down to prevent blood coagulation, and use an automatic blood cell analyzer to measure the numbers of white blood cells, lymphocytes, red blood cells, and platelets, as well as the hemoglobin content.

[0097] Tumor volume detection: After blood collection, the mice were sacrificed by cervical dislocation. The longest and shortest diameters of the tumor were measured, and the tumor volume was calculated as follows:

[0098] Tumor volume = (a × b 2 ) / 2 (where a is the longest diameter of the tumor, in mm; b is the shortest diameter of the tumor, in mm).

[0099] As shown in Table 2, after injecting CTX into the mice bearing murine lung cancer LLC cells, the numbers of white blood cells, lymphocytes, and neutrophils decreased significantly, and there were significant statistical differences compared with the blank control group (P < 0.001). However, there were no significant differences in the numbers of platelets, red blood cells, and hemoglobin (Table 3). It was considered that the model of leukopenia in tumor-bearing mice was successfully established. Compared with the tumor-bearing mice in the CTX single-treatment group (model group), the combination of low-dose Dendrobium officinale tender buds and CTX treatment (tender bud low-dose group) could significantly increase the numbers of white blood cells, lymphocytes, and neutrophils in leukopenia mice (P < 0.05). It is suggested that the Dendrobium officinale tender buds cultivated by the method disclosed in this application can significantly improve the leukopenia symptoms during the progression of chemotherapy-related myelosuppression.

[0100] Table 2. Numbers of white blood cells, lymphocytes, and neutrophils in mice of each group

[0101]

[0102]

[0103] Note: The results are expressed as mean ± standard deviation (Mean ± SD); *** indicating a significant difference P < 0.001 compared with the blank control group, # indicating a significant difference P < 0.05 compared with the model group.

[0104] Table 3. Numbers of platelets, red blood cells, and hemoglobin content in mice of each group

[0105]

[0106] Note: The results are expressed as mean ± standard deviation (Mean ± SD); # indicating a significant difference P < 0.05 compared with the model group.

[0107] The tumor volumes of mice in each group are shown in Table 4, Figure 8As shown, after 14 days of combined treatment with the tender buds of Dendrobium officinale and CTX, the average tumor volume of tumor-bearing mice was significantly reduced, and there was a significant difference compared with the treatment effect of CTX alone (P<0.001). This indicates that the tender buds of Dendrobium officinale can enhance the anti-tumor effect of the chemotherapeutic drug CTX. The tumor suppression effect of the combined treatment of Yuzhi Shengbai Tablets and CTX was significantly better than that of CTX alone (model group), comparable to the low-dose tender bud group, but weaker than the high-dose tender bud group, suggesting a dose-dependent tumor suppression effect of the tender buds of Dendrobium officinale.

[0108] Table 4. Tumor volumes of mice in each group

[0109] Group Number of animals (pcs) <![CDATA[Tumor volume (mm 3 )]]> Model group 12 295.26±19.80 Sanguisorbae Radix group for leukocytosis elevation 7 <![CDATA[110.99±30.26 *** > Low-dose tender bud group 9 <![CDATA[123.24±12.36 *** > High-dose tender bud group 10 <![CDATA[66.06±10.09 *** >

[0110] Note: The results are expressed as Mean±S.E.M; *** Indicates a significant difference P<0.001 compared with the model group.

[0111] 2.3. Detection experiment on food intake and survival period of nude mice bearing human lung cancer A549 cells

[0112] After 36 BALB / c nude mice adapted to the environment, they were randomly divided into three groups (each group was divided into two cages, with six mice in each cage): model group (0.5% CMC-Na), low-dose tender bud group (1170 mg / kg / d), and high-dose tender bud group (3510 mg / kg / d). An ectopic transplantation tumor model was constructed with human lung cancer A549 cells, and administration was carried out 24 hours after tumor bearing. The nude mice were gavaged once a day with an equal volume of 0.2 mL / 10 g body weight. During the administration process, the nude mice were given a round of CTX chemotherapy every 10 days, that is, the nude mice were intraperitoneally injected with CTX 100 mg / kg once a day. Each round of chemotherapy was continuously injected for 3 days and recovered for 7 days, and a total of 5 rounds of chemotherapy were carried out.

[0113] The method for constructing an ectopic transplantation tumor model of human lung cancer A549 cells is as follows: Collect human lung cancer A594 cells, adjust the cell number to 5×10 6 cells / mL, and inoculate 100 μL of cell suspension under the right armpit of each nude mouse to complete the construction of the transplantation tumor model of the human lung cancer A549 cell line.

[0114] Detection of nude mouse food intake: Record the construction of the ectopic transplantation tumor model of lung cancer cells as day 1, and then weigh and record the food intake of nude mice in each group every 2 days.

[0115] Detection of survival period: Check the survival status of nude mice in each group every day. After excluding accidental deaths, count the death dates of nude mice in each group and draw a survival curve.

[0116] As shown in Table 5 and Figure 9As shown in Subfigure A, compared with the model group treated with CTX alone in tumor-bearing nude mice, the low-dose tender bud group could significantly improve the average food intake of tumor-bearing nude mice during the recovery period (7 days) of each round of chemotherapy (P<0.05). As shown in Table 6 and Figure 9 As shown in Subfigure B, compared with the model group, the high-dose tender bud group could significantly improve the survival period of nude mice (P<0.01). The experimental results suggest that the Dendrobium officinale tender buds cultivated by the method disclosed in this application can effectively improve the adverse effect of reduced appetite caused by chemotherapy and can significantly extend the survival period of tumor-bearing nude mice.

[0117] Table 5. Food intake of mice in each group

[0118]

[0119]

[0120] Note: The results are expressed as Mean±S.E.M, and * indicates significant difference P<0.05 compared with the model group.

[0121] Table 6. Number of dead mice in each group

[0122]

[0123] Tumor volume detection: On the 3rd day after each round of CTX administration, the longest and shortest diameters of the tumors in nude mice were measured, and the tumor volume was calculated (the calculation method is the same as above).

[0124] As shown in Table 7 and Figure 10 As shown, after the combination treatment of Dendrobium officinale tender buds and CTX, the average tumor volume of tumor-bearing mice was significantly reduced, and there was a significant difference compared with the model group treated with CTX alone (P<0.001, P<0.05), suggesting that the Dendrobium officinale tender buds cultivated by the method disclosed in this application can enhance the anti-tumor effect of the chemotherapeutic drug CTX.

[0125] Table 7. Tumor volume of mice in each group

[0126]

[0127]

[0128] Note: The results are expressed as Mean±S.E.M, * indicates significant difference P<0.05 compared with the model group, *** indicates significant difference P<0.001 compared with the model group.

[0129] From the above experimental results, it can be seen that after treatment with the Dendrobium officinale tender buds cultivated by the method disclosed in this application, the COX-2 level in the hypothalamus of mice was significantly reduced, suggesting that this product has an inhibitory effect on fever after chemotherapy.

[0130] Further analysis showed that pre-gavage with this Dendrobium officinale young shoot product could significantly improve CTX-induced leukopenia, specifically manifested as a significant increase in the numbers of peripheral blood leukocytes, lymphocytes, and neutrophils in tumor-bearing C57BL / 6 mice; meanwhile, this product could also effectively relieve the decrease in food intake of tumor-bearing nude mice caused by CTX. The above results together indicated that this Dendrobium officinale young shoot could not only relieve chemotherapy-related fever by inhibiting the inflammatory response, but also play a role in reducing the toxicity of chemotherapy by increasing the leukocyte level and improving appetite.

[0131] In this study, by constructing two ectopic xenograft tumor models of murine lung cancer LLC cells and human lung cancer A549 cells, the inhibitory effects of CTX monotherapy and its combination with the Dendrobium officinale young shoot product on tumor growth in tumor-bearing animals were systematically evaluated. The results showed that after the combination of Dendrobium officinale young shoot and CTX treatment, the average tumor volume of tumor-bearing mice was significantly reduced compared with the CTX monotherapy group; meanwhile, this combination regimen could significantly prolong the survival period of A549 tumor-bearing nude mice. The above results indicated that Dendrobium officinale young shoot could enhance the anti-tumor activity by synergizing with CTX and play a chemotherapy-sensitizing effect.

[0132] The above shows and describes the basic principles, main features, and advantages of the present invention. However, the above are only specific embodiments of the present invention, and the technical features of the present invention are not limited thereto. Any other embodiments obtained by those skilled in the art without departing from the technical solution of the present invention should be covered within the scope of the present invention.

Claims

1. The tender buds of Dendrobium officinale with enriched active ingredients, characterized in that, It is prepared by using the tissue culture of Dendrobium officinale combined with the liquid suspension culture technology of dynamic gas regulation. The culture process goes through three stages: protocorm induction, shoot bud differentiation, and proliferation and amplification. The characteristic metabolites in the shoots of Dendrobium officinale include 2-amino-1,3,4-octadecatriol, 5-hydroxymethylfurfural, malic acid, adenosine, erucamide, 4-oxoproline, 9-hydroxy-10,12-octadecadienoic acid, and citric acid.

2. The enriched active ingredient tender Dendrobium officinale buds as described in claim 1, characterized in that, During the preparation process, a weak sugar liquid medium is used in the shoot induction and proliferation stages, and air and carbon dioxide are continuously introduced during the culture in these two stages.

3. Use of the shoots of Dendrobium officinale with enriched active ingredients as described in any one of claims 1-2 in the preparation of anti-inflammatory and antipyretic preparations.

4. Use of the tender buds of Dendrobium officinale with enriched active ingredients as described in any one of claims 1-2 in the preparation of a chemotherapy adjuvant preparation, characterized in that, The preparation has the dual functions of enhancing the efficacy of chemotherapeutic drugs and reducing their toxic and side effects.

5. The application according to claim 3, wherein The dosage form of the preparation is any one of powder, tablet, ointment, capsule, granule, syrup, pill, or oral film.

6. The application according to claim 4, characterized in that The drug delivery form of the preparation includes, but is not limited to, oral solid preparations, mucosal administration preparations, or immediate-release / sustained-release composite dosage forms.

7. A functional health product for inflammation regulation, characterized in that, It contains a functionally effective amount of the shoots of Dendrobium officinale with enriched active ingredients as described in any one of claims 1-2 as the core active component, and is jointly composed of food-grade acceptable excipients.

8. A chemotherapy synergistic adjuvant preparation, characterized in that, It contains the shoots of Dendrobium officinale with enriched active ingredients as described in any one of claims 1-2, and the components thereof achieve the directional enrichment of specific secondary metabolites through the dynamic gas regulation culture process.

9. The chemotherapy synergistic adjuvant preparation according to claim 8, wherein The auxiliary preparation is combined with a chemotherapeutic drug to form a therapeutic composition, and the chemotherapeutic drug is selected from at least one of platinum compounds, alkylating agents, and antimetabolic drugs.

10. A functional beverage, characterized in that, It contains the shoots of Dendrobium officinale with enriched active ingredients as described in any one of claims 1-2 as the active ingredient, and food-grade acceptable excipients; The drinking method of the shoots of Dendrobium officinale is to directly juice and drink the fresh shoots of Dendrobium officinale or to take them after drying and grinding the fresh shoots of Dendrobium officinale into powder.

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