Dendrobium officinale glycolipid, its preparation method and antioxidant application

Abstract

The present invention discloses dendrobium officinale glycolipid, its preparation method and antioxidant application. The dendrobium officinale glycolipid is a diglyceride rich in glucose, mannose, linoleic acid and 9,12,15-octadecatrienoic acid. The invention also discloses the alcohol extraction process of dendrobium officinale glycolipid, its applications in increasing the abundance of intestinal acid-producing bacteroides, up-regulating the expression of piceatannol and down-regulating the expression of 1-naphthol, and enhancing the activities of superoxide dismutase and glutathione peroxidase in the body, so as to ultimately improve the antioxidant capacity in vivo.

Description

Technical Field

[0001] The present invention belongs to the technical field of the research on the structure and activity of Dendrobium officinale glycolipids, and specifically relates to Dendrobium officinale glycolipids, a preparation method thereof, and an antioxidant application thereof. Background Art

[0002] Dendrobium officinale ( Dendrobium officinale ) is a perennial herbaceous plant of the genus Dendrobium in the Orchidaceae family, and is famous for its medicinal and edible properties. Modern research shows that Dendrobium officinale is rich in various bioactive components, such as polysaccharides and alkaloids, and has potential effects such as enhancing immunity, regulating blood sugar levels, and antioxidant and anti-aging effects. Generally, these active components are extracted by hot water and then separated and prepared by ethanol precipitation or column chromatography. However, the residue left after water extraction is often discarded and not effectively utilized, resulting in waste of resources. Glycolipids are complex compounds composed of water-insoluble lipids and sugars. Structurally, glycolipids mainly consist of sphingosine fatty acids as the lipid part, which are linked to monosaccharides, oligosaccharides, or polysaccharides as the sugar part through glycosidic bonds. In addition to maintaining normal cell physiological functions, glycolipids also participate in immune defense mechanisms and the development process of diseases.

[0003] During normal physiological processes, the body's metabolic activities generate reactive oxygen species (ROS). These ROS are crucial for certain cell functions (such as signal pathways and immune responses), but excessive production may cause health problems. Under normal circumstances, the body's antioxidant defense system (including superoxide dismutase, catalase, and glutathione peroxidase, etc.) can effectively neutralize excessive ROS. However, when the balance between the generation of reactive oxygen species and their clearance by antioxidant enzymes is lost, these accumulated harmful substances will damage various cell components such as proteins, lipids, and DNA, thereby destroying the normal cell structure and function and triggering a series of health problems, including inflammatory bowel disease and diabetic ulcers. Therefore, effective management of oxidative damage is crucial for maintaining optimal health and preventing / treating various diseases.

[0004] The gut microbiota refers to a group of microorganisms that inhabit the human gastrointestinal tract and coexist with the host. It not only participates in food digestion and absorption but also plays an important role in key functions such as immune regulation, metabolic control, and psychological behavior regulation. The gut microbiota can convert indigestible cellulose and polysaccharides into easily absorbable beneficial metabolites such as short-chain fatty acids, providing an energy source for cell metabolism, and at the same time has an important role in regulating immune responses, promoting colon mucosal barrier function, and inhibiting the proliferation of pathogenic microecology. In addition, some specific types of gut microbiota can produce antioxidants such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) to reduce damage caused by oxidative stress, and when the gut microbiota is imbalanced, it may lead to an increase in the level of oxidative stress.

[0005] At present, the preparation technology and bioactivity development of Dendrobium officinale glycolipids are significantly insufficient, which will limit their further application in the food and pharmaceutical fields. Summary of the Invention

[0006] The present invention provides a Dendrobium officinale glycolipid ( D. officinale glycolipid, DOG), its preparation method and antioxidant application. The Dendrobium officinale glycolipid is prepared from the residue of hot water extraction, and its structure is preliminarily analyzed. In addition, an animal model of oxidative damage is established to study its antioxidant activity based on the metabolism of intestinal flora. These research results will provide valuable data support for the further development of DOG in the field of functional foods and promote the efficient utilization of Dendrobium officinale raw materials.

[0007] To achieve the above object, the present invention provides the following technical solution: A Dendrobium officinale glycolipid, which is a diglyceride formed by glycerol (20.43%), stearic acid (1.71%), D-mannose (7.42%), D-glucose (62.36%), linoleic acid (4.38%), 9,12,15-octadecatrienoic acid (2.78%) and oleic acid (0.92%).

[0008] Preferably, the 1st and 2nd carbon atoms of glycerol in the Dendrobium officinale glycolipid are linked with sugar and fatty acid molecules.

[0009] The present invention also provides a preparation method of Dendrobium officinale glycolipid. The specific steps of the preparation method are as follows:

[0010] S1, adding the residue component after hot water extraction of Dendrobium officinale into an ethanol solution for extraction, with a liquid-to-solid ratio of 20 mL / g, an ethanol concentration of 70%, an extraction temperature of 70 °C, and an extraction time of 2.5 h to obtain a concentrated solution;

[0011] S2, treating the concentrated solution obtained in step S1 with a vacuum rotary evaporator to remove ethanol, and then performing vacuum freeze-drying to obtain Dendrobium officinale glycolipid.

[0012] The present invention also provides the application of Dendrobium officinale glycolipid in the field of antioxidant.

[0013] The application in the field of antioxidant includes being used as a dietary supplement for patients with oxidative damage, immunodeficiency, and tumors.

[0014] Compared with the prior art, the beneficial effects of the present invention are:

[0015] The Dendrobium officinale glycolipid of the present invention is a diglyceride, rich in carbohydrate components and unsaturated fatty acids, with good emulsifying properties. It can not only directly participate in the scavenging of free radicals, but also show an increase in the abundance of Bacteroides acidifaciens in the gastrointestinal tract of mice, an up-regulation of the expression of piceatannol, and a down-regulation of the expression of 1-naphthol in animal experiments, thereby enhancing the activities of SOD and GSH-Px in the serum of mice and reducing the content of MDA. MDA is malondialdehyde, and its excessive accumulation will cause cross-linking polymerization of biological macromolecules such as proteins and nucleic acids, leading to changes in the structure and function of cell membranes and belonging to group 3 carcinogens. Finally, it plays a strong antioxidant role. These results will be beneficial to the further development and utilization of Dendrobium officinale glycolipid in the fields of food and medicine.

[0016] Other features and advantages of the present disclosure will become clear from the following detailed description of the exemplary embodiments of the present disclosure with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present disclosure or related technologies, the following will briefly introduce the drawings required for use in the description of the embodiments or related technologies. Obviously, the drawings in the following description are only the embodiments of the present disclosure. For those of ordinary skill in the art, other drawings can be obtained according to the provided drawings without creative efforts.

[0018] Figure 1 It is the ultraviolet full-wavelength scanning spectrum of the Dendrobium officinale glycolipid of the present invention;

[0019] Figure 2 It is the infrared spectrum of the Dendrobium officinale glycolipid of the present invention;

[0020] Figure 3 It is the direct scavenging rate chart of the Dendrobium officinale glycolipid of the present invention against ABTS free radicals;

[0021] Figure 4 It is the direct scavenging rate chart of the Dendrobium officinale glycolipid of the present invention against DPPH free radicals;

[0022] Figure 5 It is the comparison chart of the effect of the Dendrobium officinale glycolipid of the present invention on the SOD activity in the serum of oxidized-damaged mice;

[0023] Figure 6 It is the comparison chart of the effect of the Dendrobium officinale glycolipid of the present invention on the GSH-Px activity in the serum of oxidized-damaged mice;

[0024] Figure 7 It is the comparison chart of the effect of the Dendrobium officinale glycolipid of the present invention on the MDA content in the serum of oxidized-damaged mice;

[0025] Figure 8It is a comparison chart of the effect of Dendrobium officinale glycolipid of the present invention on the total antioxidant capacity in the serum of mice with oxidative damage;

[0026] Figure 9 It is a Venn diagram of the effect of Dendrobium officinale glycolipid of the present invention on the intestinal flora of mice with oxidative damage;

[0027] Figure 10 It is a diagram of the effect of Dendrobium officinale glycolipid of the present invention on the top 10 strains of intestinal flora in mice with oxidative damage;

[0028] Figure 11 It is a ternary phase diagram of the effect of ascorbic acid on the top 10 strains of intestinal flora in mice with oxidative damage;

[0029] Figure 12 It is a ternary phase diagram of the effect of Dendrobium officinale glycolipid of the present invention on the top 10 strains of intestinal flora in mice with oxidative damage;

[0030] Figure 13 It is a heat map of the effect of ascorbic acid on the metabolites of intestinal flora in mice with oxidative damage;

[0031] Figure 14 It is a heat map of the effect of Dendrobium officinale glycolipid of the present invention on the metabolites of intestinal flora in mice with oxidative damage. Detailed implementation manners

[0032] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and in no way limits the present invention and its application or use. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0033] Please refer to Figures 1 - 14 , the present invention provides a technical solution:

[0034] The dried Dendrobium officinale is crushed into uniform powder (300 meshes), then soaked in deionized water at 80 °C for 4 h for water extraction, and the extraction is repeated 3 times. All the leachates are mixed to prepare polysaccharides. The residue after water extraction is dried and used as a raw material for the preparation of Dendrobium officinale glycolipid. Ethanol soaking and heating are carried out to improve the dissolution efficiency. The liquid-solid ratio is 20 mL / g, the ethanol concentration is 70%, the extraction temperature is 70 °C, and the extraction time is 2.5 h to obtain a concentrated solution. The concentrated solution is treated by a vacuum rotary evaporator to remove ethanol, and then vacuum freeze-dried to obtain Dendrobium officinale glycolipid (DOG). Under this process, the yield of DOG is 2.64 ± 0.18%.

[0035] The ultraviolet full-wavelength scanning spectrum of DOG was measured, and the results are as Figure 1 shown. The absorption of saturated fatty acids is relatively weak, and their absorption peaks are usually located in the lower wavelength range (200 - 210 nm). In contrast, unsaturated fatty acids have a stronger absorption capacity for ultraviolet light, and their absorption peaks are located in a higher wavelength range (220 - 230 nm). In addition, the absorption near 280 nm and 410 nm in the ultraviolet spectrum indicates the possible presence of double bonds and conjugation in DOG.

[0036] The characteristic functional groups of DOG were determined by infrared spectroscopy, and the results are as Figure 2 shown. Figure 2 The results of the infrared spectrum of -1 show that DOG contains both saturated and unsaturated fatty acids. The large absorption peak at 3439.54 cm -1 indicates its certain hydrophilicity and can be attributed to O-H stretching. The signals at 2923.37 cm -1 and 2852.05 cm -1 suggest the presence of CH2 and CH groups respectively. The absorptions at 1740.40 cm -1 and 1632.50 cm -1 belong to the stretching vibration of the carbonyl group (C=O), suggesting the presence of an ester structure. The bands appearing at 1465.68 cm -1 and 1384.11 cm -1 can be attributed to the asymmetric and symmetric bending vibrations of the CH3 group. The characteristic absorption peak at 1262.33 cm -1 mainly corresponds to the hydroxyl vibration mode in primary alcohols, while the bands found at 1165.31 cm -1 and 1076.23 cm

[0037] Table 1 shows the GC-MS analysis results of DOG after hydrolysis and acetylation. As shown in the table, the content of glycerol 1,2-diacetate in DOG is 20.43%, which indicates that the main component of DOG is diglyceride. The presence of D-glucitol hexaacetate (62.36%) and D-mannitol hexaacetate (7.42%) in DOG indicates that the sugar moiety of DOG is mainly composed of glucose and mannose. The detection of specific fatty acids such as palmitic acid (1.71%), linoleic acid (4.38%), 9,12,15-octadecatrienoic acid (2.78%), and oleic acid (0.92%) is consistent with the previous detection results. The structural framework of glyceroglycolipids is composed of a glycerol backbone covalently linked to monosaccharide residues and fatty acids through O-glycosidic bonds, widely existing in the plant, animal, and bacterial kingdoms, participating in processes such as intercellular communication, immune response regulation, and photosynthetic electron transfer, and having antioxidant potential.

[0038] Table 1 Monomer Composition of DOG

[0039] Serial number Compound description Molecular formula Retention time (min) Molecular weight (Da) Ratio (%) 1 Glycerol 1,2 - diacetate <![CDATA[C7H 12 O5]]> 4.79 176.07 20.43% 2 Ethyl stearate <![CDATA[C 18 H 36 O2]]> 10.55 284.27 1.71% 3 D - Mannitol, hexaacetate <![CDATA[C 18 H 26 O 12 > 12.45 434.14 7.42% 4 D - Glucitol, hexaacetate <![CDATA[C 18 H 26 O 12 > 12.66 434.14 62.36% 5 Ethyl linoleate <![CDATA[C 20 H 36 O2]]> 13.02 308.27 4.38% 6 9,12,15 - Octadecatrienoic acid, ethyl ester <![CDATA[C 20 H 34 O2]]> 13.13 306.26 2.78% 7 Vaccenic acid, ethyl ester <![CDATA[C 20 H 40 O2]]> 13.42 312.30 0.92%

[0040] Figure 3 and Figure 4 shows the scavenging rates of DOG on ABTS and DPPH free radicals, with ascorbic acid as the positive control. ABTS free radical refers to: when a substance is added to the ABTS free radical solution, if the absorbance at 734 nm decreases, it indicates that the substance has free radical scavenging activity and belongs to an antioxidant. This method is called the ABTS free radical scavenging method and can be used to evaluate the antioxidant capacity of plants (or Chinese herbal medicine extracts) and pure compounds. DPPH free radical is a stable free radical and is widely used to quantitatively determine the in vitro antioxidant capacity of biological samples, pure compounds, and extracts. As shown in the figure, as the concentrations of DOG and ascorbic acid increase from 0.25 to 4.00 mg / mL, their scavenging effects on ABTS and DPPH free radicals show a similar trend. Ascorbic acid has strong antioxidant activity in vitro. Even at the lowest concentration (0.25 mg / mL), it can achieve a free radical scavenging ability of about 90%, and further increasing the concentration cannot continue to significantly improve its antioxidant activity. However, as the concentration of DOG increases, the average scavenging rate of ABTS free radicals increases from 30.97% to 88.78%, and the average scavenging rate of DPPH free radicals increases from 39.56% to 83.09%, indicating that DOG also has a certain ability to directly scavenge free radicals.

[0041] Purchase 50 six-week-old Kunming mice (weighing 25 ± 2 g) from Jinan Pengyue Experimental Animal Breeding Co., Ltd. The mice are raised in an environment with a relative humidity of 45% - 55% and a temperature controlled at 20 - 25 °C. After the adaptation period, the mice are randomly divided into five groups (10 mice in each group): blank group, model group, ascorbic acid group, low-dose DOG treatment group (DOG-L, 50 mg / kg), and high-dose DOG treatment group (DOG-H, 100 mg / kg). The specific experimental procedure is as follows: At the beginning of the experiment, the blank group and the model group are intragastrically administered 0.2 mL of normal saline (0.9%) daily, while the ascorbic acid group is intragastrically administered 50 mg / kg of ascorbic acid, and the DOG-L and DOG-H groups are respectively given the corresponding doses of DOG for 21 days. On the 8th day, except for the blank group, the mice in each experimental group are injected with 60 mg / kg of cyclophosphamide (CTX) for 3 days. On the 22nd day, the serum and fecal samples of the mice are collected for further analysis.

[0042] Figure 5 and Figure 6The SOD and GSH-Px activities in the sera of mice in each group were shown respectively. As shown in the figure, compared with the blank group, the activities of SOD and GSH-Px in the sera of mice in the model group were significantly decreased ( p <0.01), indicating that intraperitoneal injection of cyclophosphamide caused severe damage to the body's antioxidant system. After ascorbic acid intervention, no significant improvement was observed, indicating that ascorbic acid could not effectively enhance the antioxidant enzyme activity in vivo. On the contrary, after intragastric administration of DOG, the activities of these two antioxidant enzymes in the sera of mice increased to varying degrees, indicating that DOG could improve the overall antioxidant level by increasing the antioxidant enzyme activity.

[0043] Figure 7 The content of MDA in the sera of mice in each group was shown. As shown in the figure, compared with the blank group, the content of MDA in the sera of mice in the model group was significantly increased ( p <0.01), suggesting that cyclophosphamide inhibited the body's antioxidant system. After treatment with ascorbic acid and DOG, the content of MDA in the sera of mice in the model group decreased significantly, indicating that they had certain antioxidant effects in vivo, but their action mechanisms might be different. DOG exerted its antioxidant function in vivo by enhancing the activity of related enzymes, while ascorbic acid achieved its antioxidant effect by directly scavenging free radicals.

[0044] Figure 8 The direct scavenging effect of the sera of mice in each group on free radicals was shown. The results showed that there was no significant difference in the direct scavenging ability of the sera of mice in each group on free radicals, indicating that the peripheral blood of the body had good self-coordination ability.

[0045] Figure 9 The changes in the intestinal flora abundances of mice in each group were shown. As shown in the figure, there were 461 common operational taxonomic units (OTUs) in these groups, and the specific OTUs in the blank group, model group, ascorbic acid group, and DOG group were 121, 137, 69, and 124 respectively, indicating that various intervention treatments had obvious effects on the intestinal microbial diversity of mice with oxidative damage.

[0046] Figure 10Shows the changes in the top 10 gut bacterial species content in each group of mice. As shown in the figure, compared with the blank group, the contents of Clostridium papyrosolvens, Bacteroides vulgatus, and Helicobacter hepaticus in the model group increased significantly, while the contents of Escherichia coli and Bacteroides acidifaciens decreased significantly. This indicates that colitis occurred in the cyclophosphamide-treated mice, increasing the oxidative stress response. Compared with the model group, the contents of Escherichia coli and Helicobacter MIT01-6451 in the ascorbic acid group of mice increased significantly, while the contents of Clostridium papyrosolvens and Bacteroides vulgatus decreased. At the same time, the DOG group had higher contents of Helicobacter MIT 01-6451 and Bacteroides acidifaciens, and lower proportions of Bacteroides vulgatus and Clostridium papyrosolvens. These results suggest that the antioxidant effects of DOG and ascorbic acid are different.

[0047] Figure 11 and Figure 12 Displays the ternary phase diagrams of the top 10 gut bacterial species contents in the blank-group - model-group - ascorbic acid group and blank-group - model-group - DOG group. As shown in the figure, it can be seen more intuitively that Clostridium papyrosolvens and Bacteroides vulgatus in the model group mainly act by inhibiting the body's antioxidant system, while Helicobacter MIT 01-6451 in the ascorbic acid group and DOG group may reflect the antioxidant and immune regulation levels in the body. Escherichia coli mainly plays an important indicator role in the oxidatively damaged mice after ascorbic acid intervention, while Bacteroides acidifaciens can enhance the antioxidant activity induced by DOG by protecting liver metabolism. This is consistent with the previous analysis results.

[0048] Table 2 shows the information on the gut differential metabolites in the ascorbic acid group and model group of mice. The results show that there are 25 metabolites with significantly different expressions, among which 15 are down-regulated and 10 are up-regulated.

[0049] Table 2 Differential expression of metabolites in the ascorbic acid group compared with the model group

[0050] Metabolite ID Compound description Molecular formula Retention time (min) P - value Expression C879 PG(18:3(9Z,12Z,15Z) / 18:3(6Z,9Z,12Z)) <![CDATA[C 42 H 71 O 10 P]]> 4.67 0.0192 Down - regulated C271 8 - Isoprostaglandin F2a <![CDATA[C 20 H 34 O5]]> 5.55 0.0341 Down - regulated C287 ACar 15:1 <![CDATA[C 22 H 42 NO4]]> 8.19 0.0194 Down - regulated C897 Progesterone <![CDATA[C 21 H 30 O2]]> 6.76 0.0235 Down - regulated C959 Urobilin <![CDATA[C 33 H 46 N4O6]]> 10.18 0.0338 Down - regulated C298 ACar 20:2 <![CDATA[C 27 H 50 NO4]]> 9.36 0.0262 Down - regulated C820 PC (22:6e / 18:5) <![CDATA[C 48 H 76 NO7P]]> 7.68 0.0343 Down - regulated C711 N - Propionyl - phenylalanine <![CDATA[C 12 H 15 NO4]]> 1.17 0.0179 Down - regulated C106 2 - (1H - 1,2,3 - Benzotriazol - 1 - yl) - N - (2,3 - dihydro - 1H - inden - 2 - yl) acetamide <![CDATA[C 17 H 16 N 4 O]]> 13.14 0.0305 Down - regulated C437 DL - o - Tyrosine <![CDATA[C9H 11 NO3]]> 2.87 0.0239 Down - regulated C936 SM(d18:0 / 16:0) <![CDATA[C 39 H 81 N2O6P]]> 4.67 0.0459 Down - regulated C672 N,N - Dimethylsphing - 4 - malvidin - 3 - glucoside <![CDATA[C 20 H 41 NO2]]> 9.15 0.0041 Down - regulated C666 N - (3 - Oxohexanoyl) homoserine lactone <![CDATA[C 10 H 15 NO4]]> 9.18 0.0372 Down - regulated C362 Ceramide(d18:1 / 16:0) <![CDATA[C 34 H 67 NO3]]> 10.06 0.0159 Down - regulated C64 Ketoetiocholanolone <![CDATA[C 19 H 28 O3]]> 13.87 0.0444 Down - regulated C673 Dihydrocapsaicin <![CDATA[C 18 H 29 NO3]]> 13.72 0.0057 Up - regulated C548 L - 2 - Amino - 3 - oxobutyric acid <![CDATA[C4H7NO3]]> 0.56 0.0174 Up - regulated C236 5,7 - Dimethyl - 2 - phenylpyrazolo[1,5 - a]pyrimidine <![CDATA[C 14 H 13 N3]]> 13.24 0.0223 Up - regulated C320 All - trans 13,14 - Dihydroretinol <![CDATA[C 20 H 32 O]]> 5.11 0.0362 Up - regulated C384 Creatinine <![CDATA[C4H7N3O]]> 0.57 0.0274 Up - regulated C246 5 - Methoxyindoleacetic acid <![CDATA[C 11 H 11 NO3]]> 4.06 0.0185 Up - regulated C615 Lysophosphatidylcholine(14:0) <![CDATA[C 22 H 46 NO7P]]> 6.51 0.0340 Up - regulated C385 Cryptotanshinone <![CDATA[C 19 H 20 O3]]> 4.3 0.0102 Up - regulated C118 2,4 - Dihydroxybenzoic acid <![CDATA[C7H6O4]]> 0.83 0.0004 Up - regulated C192 3 - Methoxycinnamic acid <![CDATA[C 10 H 10 O3]]> 4.27 0.0126 Up - regulated

[0051] Figure 13 Performed a heatmap analysis of these metabolites in a more intuitive way. Among Figure 13In the figure, the color gradient from blue to red indicates the increase in metabolite concentration, making the expression differences between different metabolites clearer. In addition, through the signal pathway enrichment analysis of these metabolites, two significantly changed pathways were found: progesterone (C897)-mediated oocyte meiosis and maturation. Studies have shown that progesterone can promote the expression of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPX) in cells, especially oocytes, and improve their antioxidant activity through related signal pathways. However, the results of this study showed that ascorbic acid intervention led to a significant downregulation of progesterone levels in oxidative stress mice, indicating that the signal pathway mediated by progesterone was also inhibited, which is consistent with previous studies that ascorbic acid cannot enhance the activity of antioxidant enzymes.

[0052] Table 3 shows the information of differential metabolites in the intestine of mice in the DOG group and the model group. The results showed that a total of 63 metabolites showed significant differences, of which 35 were downregulated and 28 were upregulated.

[0053] Table 3 Differential expression of metabolites in the DOG group compared with the model group

[0054] Metabolite ID Compound description Molecular formula Retention time (min) P - value Expression C1014 (2,3,9,17,22R) - 2,3,14,20,22 - Pentahydroxyergost - 7 - en - 6 - one <![CDATA[C 28 H 46 O6]]> 6.28 0.0025 Down - regulated C576 LPC 14:0 <![CDATA[C 22 H 46 NO7P]]> 8.60 0.0019 Down - regulated C97 1 - Naphthol <![CDATA[C 10 H8O]]> 13.00 0.0080 Down - regulated C537 Inosine <![CDATA[C 10 H 12 N4O5]]> 4.05 0.0006 Down - regulated C108 2 - (1H - Indol - 3 - yl) acetic acid <![CDATA[C 10 H9NO2]]> 15.19 0.0085 Down - regulated C92 1 - Methyladenosine <![CDATA[C 11 H 15 N5O4]]> 0.55 0.0034 Down - regulated C959 Urobilin <![CDATA[C 33 H 46 N4O6]]> 10.18 0.0182 Down - regulated C480 FRH <![CDATA[C 21 H 30 N8O4]]> 10.88 0.0435 Down - regulated C179 3 - Hydroxy - 5, 8 - carnitine <![CDATA[C 19 H 25 BN4O4]]> 6.17 0.0366 Down - regulated C809 PC (19:0 / 19:1) <![CDATA[C 46 H 90 NO8P]]> 10.45 0.0359 Down - regulated C840 PC(18:1(11Z) / 20:0) <![CDATA[C 46 H 90 NO8P]]> 13.30 0.0341 Down - regulated C287 ACar 15:1 <![CDATA[C 22 H 42 NO4]]> 8.19 0.0350 Down - regulated C1105 (13E,16E,19E) - Docosatri - 13,16,19 - enolic acid <![CDATA[C 22 H 38 O2]]> 9.62 0.0286 Down - regulated C188 3 - Indoleacrylic acid <![CDATA[C 11 H9NO2]]> 3.22 0.0251 Down - regulated C437 DL - o - Tyrosine <![CDATA[C9H 11 NO3]]> 2.87 0.0157 Down - regulated C282 ACar 12:1 <![CDATA[C 19 H 36 NO4]]> 5.93 0.0184 Down - regulated C883 Choline phosphate <![CDATA[C5H 14 NO4P]]> 13.19 0.0348 Down - regulated C1147 PC (18:2e / 20:4) <![CDATA[C 46 H 82 NO7P]]> 13.17 0.0152 Down - regulated C823 PC (9:0 / 9:0) <![CDATA[C 26 H 52 NO8P]]> 8.43 0.0390 Down - regulated C906 Protectin D1 <![CDATA[C 22 H 32 O4]]> 6.73 0.0224 Down - regulated C699 N - Arachidonoyl - L - serine <![CDATA[C 23 H 37 NO4]]> 13.02 0.0420 Down - regulated C112 2 - (3,5 - Dimethyl - 1H - pyrazol - 4 - yl) - 5 - methoxybenzoic acid <![CDATA[C 13 H 14 N 2 O3 > 4.05 0.0314 Down - regulated C578 LPC 15:0 <![CDATA[C 23 H 48 NO7P]]> 6.87 0.0168 Downregulation C711 N-propionyl-phenylalanine <![CDATA[C 12 H 15 NO4]]> 1.17 0.0013 Downregulation C1103 (+ / -)11(12)-EET <![CDATA[C 20 H 32 O3]]> 7.44 0.0266 Downregulation C1000 Uridine <![CDATA[C9H 12 N2O6]]> 0.58 0.0339 Downregulation C541 Cafestol <![CDATA[C 20 H 26 O3]]> 2.98 0.0397 Downregulation C1115 2-Arachidonoylglycerol <![CDATA[C 23 H 38 O4]]> 9.03 0.0009 Downregulation C776 PC (17:1 / 17:2) <![CDATA[C 42 H 78 NO8P]]> 9.77 0.0010 Downregulation C1109 12-HETE <![CDATA[C 20 H 32 O3]]> 7.26 0.0328 Downregulation C449 D-Phenylalanine <![CDATA[C9H 11 NO2]]> 1.43 0.0202 Downregulation C452 Ecgonine <![CDATA[C9H 15 NO3]]> 3.25 0.0166 Downregulation C64 11-Ketotestosterone <![CDATA[C 19 H 28 O3]]> 13.87 0.0366 Downregulation C534 Indoline-2-carboxylic acid <![CDATA[C9H9NO2]]> 1.35 0.0391 Downregulation C888 Pimelyl carnitine <![CDATA[C 14 H 25 NO6]]> 5.25 0.0136 Downregulation C279 9-Oxo-ODE <![CDATA[C 18 H 30 O3]]> 6.44 0.0136 Upregulation C28 (9Z)-(7S,8S)-Dihydroxyoctadecadienoate <![CDATA[C 18 H 34 O4]]> 6.07 0.0136 Upregulation C1021 12(13)-DiHOME <![CDATA[C 18 H 34 O4]]> 9.23 0.0147 Upregulation C115 2,3-Dihydroxypropyl-12-methyltridecanoate <![CDATA[C 17 H 34 O4]]> 6.79 0.0192 Upregulation C446 Lauroyl carnitine <![CDATA[C 19 H 37 NO4]]> 7.78 0.0353 Upregulation C320 All-trans 13,14-dihydroretinol <![CDATA[C 20 H 32 O]]> 5.11 0.0136 Upregulation C23 (6R,7S)-6,7-Epoxyoctadecanoic acid <![CDATA[C 16 H 26 O5]]> 7.41 0.0136 Upregulation C595 LPE 18:2 <![CDATA[C 23 H 44 NO7P]]> 6.93 0.0462 Upregulation C1 (+ / -)12(13)-DiHOME <![CDATA[C 18 H 34 O4]]> 6.47 0.0092 Upregulation C316 Adrenic acid <![CDATA[C 22 H 36 O2]]> 0.59 0.0087 Upregulation C12 (2R)-(9Z,12Z,15Z)-2-Hydroperoxyoctadecatri-9,12,15-enoic acid <![CDATA[C 20 H 40 O2]]> 6.03 0.0489 Upregulation C1129 D-(+)-Maltose <![CDATA[C 12 H 22 O 11 > 4.32 0.0249 Upregulation C236 5,7-Dimethyl-2-phenylpyrazolo[1,5-a]pyrimidine <![CDATA[C 14 H 13 N3]]> 13.24 0.0259 Upregulation C283 ACar 12:3 <![CDATA[C 19 H 32 NO4]]> 5.27 0.0004 Upregulation C277 9-HpODE <![CDATA[C 18 H 32 O4]]> 6.5 0.0354 Upregulation C926 SM (d17:0 / 23:1) <![CDATA[C 45 H 91 N2O6P]]> 3.38 0.0015 Upregulation C228 4-Cresol <![CDATA[C7H8O]]> 3.69 0.0094 Upregulation C1123 9-HpODE <![CDATA[C 18 H 32 O4]]> 6.53 0.0270 Upregulation C24 (7S,8S)-DiHODE <![CDATA[C 18 H 32 O4]]> 6.44 0.0261 Upregulation C599 LPE 20:5 <![CDATA[C 25 H 42 NO7P]]> 6.93 0.0409 Upregulation C652 Matricin <![CDATA[C 17 H 22 O5]]> 5.43 0.0290 Upregulation C493 Ginkgolic acid <![CDATA[C 22 H 34 O3]]> 8.89 0.0307 Upregulation C294 ACar 18:2 <![CDATA[C 25 H 46 NO4]]> 7.41 0.0269 Upregulation C4 (11E,15Z)-9,10,13-Trihydroxyoctadeca-11,15-dienoic acid <![CDATA[C 18 H 32 O5]]> 4.78 0.0121 Upregulation C319 All-cis-4,7,10,13,16-Docosapentaenoic acid <![CDATA[C 22 H 34 O2]]> 6.47 0.0033 Upregulation C994 Quinodimethacrylate <![CDATA[C 14 H 18 O4]]> 5.78 0.0445 Upregulation C758 PC (16:0 / 17:0) <![CDATA[C 41 H 82 NO8P]]> 8.25 0.0358 Upregulation C1167 Piceatannol <![CDATA[C 14 H 12 O4]]> 0.58 0.0204 Upregulation

[0055] Figure 14 A heat map analysis of these metabolites was performed in a more intuitive way. Figure 14 In the figure, the color gradient from blue to red indicates the increase in metabolite concentration, making the expression differences between different metabolites clearer. In addition, through the signal pathway enrichment analysis of these metabolites, four pathways were found: 4-methylphenol (C228)-mediated nitrotoluene degradation, piceatannol (C1167)-mediated flavonoids, diphenylheptane compounds and turmeric ketones, and 1-naphthol (C97)-mediated cytochrome P450 and naphthalene family metabolism of exogenous substances. In this application, DOG mainly exerts its antioxidant effect by enhancing the activity of related enzymes in the body, reducing the potential direct free radical scavenging effect of 4-methylphenol. Studies have shown that some microorganisms can synthesize piceatannol during fermentation and enhance the activity of antioxidant enzymes in the body. In contrast, 1-naphthol and its metabolites can induce the massive production of ROS in the body, destroying the inherent redox balance and triggering oxidative stress. Therefore, the antioxidant mechanism of DOG may be related to upregulating the expression of piceatannol and downregulating the expression of 1-naphthol, thereby increasing the activity of antioxidant enzymes and ultimately exerting a protective effect against cyclophosphamide-induced oxidative damage.

[0056] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A Dendrobium officinale glycolipid, characterized in that: The Dendrobium officinale sugar lipid is a diglyceride formed by 20.43% of 1,2-diacetate of glycerol, 1.71% of stearic acid, 7.42% of D-mannose, 62.36% of D-glucose, 4.38% of linoleic acid, 2.78% of 9,12,15-octadecatrienoic acid and 0.92% of octadecenoic acid.

2. The method for preparing a Dendrobium officinale glycolipid according to claim 1, characterized in that: The specific steps of the preparation method are as follows: S1, adding the residue components after hot water extraction of Dendrobium officinale to ethanol solution for extraction, with a liquid-to-solid ratio of 20 mL / g, an ethanol concentration of 70%, an extraction temperature of 70°C, and an extraction time of 2.5 h to obtain a concentrated solution; S2, the concentrated solution obtained in step S1 is treated by a vacuum rotary evaporator to remove ethanol, and then subjected to vacuum freeze-drying to obtain Dendrobium officinale glycolipid.

3. Use of the Dendrobium officinale glycolipid as claimed in claim 1 in the preparation of antioxidant drugs.

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