Cinnamomum camphora polysaccharide as well as extraction method and application thereof

Abstract

The invention relates to an extraction method of Cinnamomum camphora polysaccharide, which comprises the following steps: crushing Cinnamomum camphora fruits, heating and extracting with water at 65-90 DEG C, concentrating a water extracting solution under reduced pressure, carrying out alcohol precipitation on a concentrated solution, deproteinizing, carrying out alcohol precipitation again, and freeze-drying to obtain the Cinnamomum camphora polysaccharide A; or crushing Cinnamomum camphora fruits, respectively extracting the crushed Cinnamomum camphora fruits with petroleum ether and ethyl acetate at room temperature, carrying out solid-liquid separation, carrying out heating extraction on residues with water at 65-90 DEG C, carrying out vacuum concentration on a water extract, carrying out alcohol precipitation on a concentrated solution, deproteinizing, carrying out alcohol precipitation again, and freeze-drying to obtain Cinnamomum camphora polysaccharide B; the Cinnamomum camphora polysaccharide A and / or the Cinnamomum camphora polysaccharide B are / is the Cinnamomum camphora polysaccharide. The invention also discovers a new application of the camellia oleifera tree polysaccharide, and the extracted camellia oleifera tree polysaccharide has a remarkable treatment effect and no toxic or side effect, and can be used for development and utilization of anti-oxidative stress drugs, especially liver protection drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of medicine and / or health products, and specifically relates to a tea fruit camphor polysaccharide, an extraction method and application thereof in preparing an antioxidant stress-resistant drug, especially a liver protection drug. Background Art

[0002] Under normal circumstances, the antioxidant-oxidative system is in a dynamic equilibrium state in the body, but when free radicals accumulate excessively, they will cause oxidative damage to proteins, lipids and nucleic acids. With the abuse of synthetic antioxidants and the emergence of side effects, people are increasingly paying attention to finding effective, natural antioxidants to reduce the damage of free radicals and antioxidants to our bodies.

[0003] As an important metabolic organ, the liver actively participates in various life processes and energy metabolism in the human body, making it extremely vulnerable to damage from various endogenous or exogenous substances. Exogenous substances such as drugs, infections, alcohol, and chemicals may induce liver poisoning, leading to an overall decline in liver metabolic function. Carbon tetrachloride (CCl4) is a classic exogenously induced hepatotoxic substance. An increasing number of studies have shown that oxidative stress is an important mechanism of CCl4-induced hepatotoxicity. CCl4 is metabolized by the cytochrome P450 system to form trichloromethyl free radicals (CCl3-) and trichloromethyl peroxyl free radicals (CCl3OO-), thereby inducing membrane lipid peroxidation.

[0004] Tea fruit camphor ( Cinnamon chago BSSun et HLZhao) is a genus of the Lauraceae family (Cinnamomum spp. Cinnamomum ). Cinnamomum camphora often grows on hillside roadsides and in shrub forests. The wood is fragrant, the fruit is medium-sized berry-shaped, the kernel is rich in oil, and the fruit can be eaten after peeling, drying or frying. Local people in the distribution area call it "tea fruit". Tea fruit is not only delicious, but also has the effect of clearing away heat and improving eyesight. However, there are no reports on the activity research of Cinnamomum camphora polysaccharides. It is of certain research significance to explore the liver protective effect of Cinnamomum camphora. Based on this, this application was developed. Summary of the invention

[0005] The purpose of the present invention is to overcome the defects of the prior art and provide a tea fruit cinnamon polysaccharide. The prepared tea fruit cinnamon polysaccharide has significant therapeutic effects and no toxic side effects. It can be used for the development and utilization of anti-oxidative stress drugs, especially liver protection drugs. That is, the present invention has discovered a new use of tea fruit cinnamon polysaccharide.

[0006] The present invention also provides a method for extracting the tea fruit camphor polysaccharide and its application in preparing anti-oxidative stress drugs, especially liver protection drugs.

[0007] To achieve the above object, the present invention adopts the following technical solution: A method for extracting tea fruit cinnamon polysaccharide, comprising the following steps: The cinnamomum camphora fruit is crushed and then heated with water at 65-90°C for extraction, the water extract is concentrated under reduced pressure, the concentrate is precipitated with alcohol, protein is removed, and then precipitated with alcohol again, and freeze-dried to obtain cinnamomum camphora polysaccharide A; Alternatively, after the fruits of Cinnamomum camphora are crushed, they are extracted with petroleum ether and ethyl acetate at room temperature, the solid and liquid are separated, the residue is extracted with water at 65-90°C, the water extract is concentrated under reduced pressure, the concentrate is precipitated with alcohol, the protein is removed, and then precipitated with alcohol again, and freeze-dried to obtain Cinnamomum camphora polysaccharide B; The tea fruit cinnamon polysaccharide A and / or tea fruit cinnamon polysaccharide B are tea fruit cinnamon polysaccharides.

[0008] Specifically, water can be added according to a solid-liquid ratio of 1 g: 20-40 ml for heating extraction, the heating extraction time is 2-5 hours, and the heating extraction is performed 1-4 times.

[0009] Furthermore, petroleum ether or ethyl acetate can be added for leaching at a solid-liquid ratio of 1 g: 10-30 ml, the leaching time is 8-15 h, and the leaching is performed 1-5 times.

[0010] In the extraction method of the present invention, after the concentrated solution is precipitated with alcohol and the protein is removed, the alcohol precipitation can be performed again using conventional techniques in the art, such as the following operation: The concentrate was added with ethanol to a final concentration of 75±5%, allowed to stand, and the precipitate was collected. After being dissolved in water, the protein was removed by the Sevage method. The supernatant was added with ethanol to a final concentration of 75±5%, allowed to stand, and the precipitate was freeze-dried.

[0011] The invention provides tea fruit cinnamomum camphora polysaccharide extracted by the extraction method.

[0012] The present invention provides a compound preparation comprising the above-mentioned cinnamomum camphora polysaccharide, wherein the cinnamomum camphora polysaccharide is compounded with conventional auxiliary materials in the art to prepare the compound preparation.

[0013] Furthermore, the dosage form of the compound preparation includes but is not limited to tablets, granules, pills, capsules or injections.

[0014] The present invention provides the use of the tea fruit camphor polysaccharide or the compound preparation in preparing a medicine for treating or preventing oxidative stress.

[0015] Furthermore, the oxidative stress is oxidative stress caused by cyclophosphamide; the camphor polysaccharide or compound preparation can effectively reduce the malondialdehyde (MDA) content of the liver caused by oxidative stress caused by cyclophosphamide, increase the levels of catalase (CAT) and total antioxidant capacity (T-AOC) in the liver, effectively enhance the antioxidant capacity, and have a good protective effect on liver damage.

[0016] The present invention also provides the use of the tea fruit camphor polysaccharide or the compound preparation in preparing a drug for treating or preventing liver damage, which can also be said to be an application in preparing a liver protection drug.

[0017] Furthermore, the liver injury is acute liver injury; the camphor tree polysaccharide or compound preparation can improve the liver function indicators of Kunming mice, significantly reduce the levels of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase, and at the same time can significantly alleviate the damage to the liver organ and effectively enhance the body's antioxidant capacity.

[0018] Compared with the prior art, the present invention has the following beneficial effects: The present invention uses tea fruit camphor polysaccharide to administer to mice by gavage once a day, and injects 0.3% carbon tetrachloride olive oil solution after three weeks to model and establish a mouse acute liver injury model. Based on the above conditions, a new use of tea fruit camphor polysaccharide is found. The research results show that compared with the model group, tea fruit camphor polysaccharide can significantly reduce the activity of mouse aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase, and enhance the antioxidant capacity of mice. At the same time, it can significantly alleviate the damage of mouse liver organs and play a liver protection effect. Therefore, the tea fruit camphor polysaccharide of the present invention can be used to prepare liver protection drugs, and has the potential and value of developing new liver protection drugs and health products. In addition, tea fruit camphor polysaccharide can also significantly reduce the malondialdehyde (MDA) content of mouse liver, increase the level of catalase (CAT) and total antioxidant capacity (T-AOC) in mouse liver, and enhance the antioxidant capacity of mice. Therefore, the tea fruit camphor polysaccharide of the present invention can be used to prepare drugs for resisting oxidative stress caused by cyclophosphamide, and has the potential and value of developing new drugs and health products for resisting oxidative stress. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The effect of Cinnamomum camphora polysaccharide on mouse liver tissue morphology, BC is the blank group, MC is the model group, PC is the bifendate positive control group, HD, MD, LD are the high, medium and low dose groups of Cinnamomum camphora polysaccharide, the same below; Figure 2 The effect of Cinnamomum camphora polysaccharide on the activities of aspartate aminotransferase (AST) (A), alanine aminotransferase (ALT) (B), and alkaline phosphatase (AKP) (C) in mouse serum. The data are expressed as mean ± standard deviation, n = 10. Compared with the blank group, ### P <0.001, ## P <0.01, # P ﹤0.05. Compared with the model group,*** P <0.001,** P <0.01,* P ﹤0.05. Same below; Figure 3 The effects of Cinnamomum camphora polysaccharide on superoxide dismutase (SOD) activity (A), glutathione peroxidase (GSH-Px) activity (B) and malondialdehyde (MDA) content (C) in mouse liver. Figure 4 Effects of Cinnamomum camphora polysaccharide on CAT (A) activity, T-AOC (B) activity and MDA (C) activity in mouse liver (data are expressed as mean ± SD, n = 8. Compared with the blank group, ### P <0.001, ## P <0.01, # P ﹤0.05. Compared with the model group,*** P <0.001,** P <0.01,* P ﹤0.05). DETAILED DESCRIPTION

[0020] The technical solution of the present invention is further described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0021] Tea fruit camphor ( Cinnamomum chago BS Sun & H. L. Zhao) fruits were collected from Yangbi County, Yunnan Province. In the following examples, ethanol concentrations are all referred to as volume percentage concentrations unless otherwise specified.

[0022] Example 1 Preparation of Cinnamomum camphora polysaccharide: A method for preparing tea fruit cinnamon polysaccharide comprises the following steps: 5342 g of Cinnamomum camphora fruit were crushed, and water was added at a solid-liquid ratio of 1g:30 ml for heating extraction at 80°C. The heating extraction time was 4 h each time, and the extraction was repeated twice. After the water extracts were combined, they were concentrated under reduced pressure at below 60°C using a rotary evaporator. 95% ethanol was added to the concentrate until the final ethanol concentration was 75%, and the solution was allowed to stand for 24 h. The solution was centrifuged and the precipitate was collected. The precipitate was dissolved in water and the protein was removed by the Sevage method (conventional techniques in the art can be used, or as follows: sample: Sevage reagent = 15:5, Sevage reagent is a chloroform-n-butanol mixture with a volume ratio of 4:1; oscillate for 60 min, stand for 30 min, centrifuge at 7000 r / min for 6 min, and remove the denatured protein outside the interface between the water layer and the chloroform layer. The upper sugar solution was taken and the operation of removing the denatured protein was repeated until all the protein was removed). The supernatant was added with 95% ethanol to a final ethanol concentration of 75%, and allowed to stand for 24 h. The precipitate was collected and freeze-dried (-50 °C, 12 h) to obtain 31.7 g of cinnamomum camphora polysaccharide with a yield of about 0.59% for later use.

[0023] The protective effect of the cinnamomum camphora polysaccharide prepared in Example 1 on mice with acute liver injury induced by carbon tetrachloride was investigated below.

[0024] Application experiment 1 Animal experiment on acute liver injury caused by carbon tetrachloride in mice: The tea fruit camphor polysaccharide prepared in Example 1 was subjected to in vivo animal experiments. 1. Experimental Animals Kunming (KM) mice, Specific pathogen free (SPF) grade, male, 5-6 weeks old, 22-28 g, were purchased from Henan Sikebes Biotechnology Co., Ltd. The mice were adaptively raised for 1 week at a temperature of 25±2°C, a relative humidity of 40-45%, and a light intensity of 12 h / d, and were fed with standard feed and had free access to water.

[0025] 2. Test reagents and instruments Electronic balance (ME204, METTLER TOLEDO); Rotary evaporator (N-1300, EYELA); FMB40 fully automatic flake ice machine (Shanghai Bilang Instrument Co., Ltd.); Desktop centrifuge, MiniSpin, Shanghai Anting Scientific Instrument Factory; Full wavelength microplate reader, 1510 Thermo Fisher; KZ-Ⅲ-FP high-speed cryogenic tissue grinder (Wuhan Saiweier Biotechnology Co., Ltd.); Electric heating blast drying oven (Shanghai Yiheng Scientific Instrument Co., Ltd.); Carbon tetrachloride (CCl4, analytical grade) (Tianjin Hongyan Chemical Reagent Factory); Bifendate dripping pills (Zhejiang Wanbang Pharmaceutical Co., Ltd.); Aspartate aminotransferase (AST / GOT) kit (Nanjing Jiancheng Bioengineering Institute); Alanine aminotransferase (ALT / GPT) kit (Nanjing Jiancheng Bioengineering Institute); Alkaline phosphatase (AKP) kit (Nanjing Jiancheng Bioengineering Institute); Malondialdehyde (MDA) assay kit (Nanjing Jiancheng Bioengineering Institute); Superoxide dismutase (SOD) assay kit (Nanjing Jiancheng Bioengineering Institute); Glutathione peroxidase (GSH-Px) assay kit (Nanjing Jiancheng Bioengineering Institute); BCA protein concentration determination kit (Beijing Solebow Technology Co., Ltd.).

[0026] 3. Test methods Table 1 Experimental groups and corresponding dosages After 1 week of adaptive feeding, the mice were randomly divided into 6 groups: blank group, model group, positive drug group, low-dose group, medium-dose group, and high-dose group, with 10 mice in each group. The corresponding dosages are shown in Table 1. Each group of mice was gavaged once a day at a volume of 10 mL / kg for 3 weeks. All drugs were dissolved in normal saline for administration.

[0027] The mice were weighed every 2 days, and the mice were gavaged daily. The volume of each gavage was adjusted according to the weight changes of the mice. The blank group and the model group were given normal saline, the positive drug PC group was given bifendate, and the drug group was given tea fruit camphor polysaccharide. 12 hours after the experimental samples were given on the last day of the experiment, except for the blank group (the blank group mice were intraperitoneally injected with olive oil at a volume of 10 mL / kg), the other groups of mice were injected with 0.3% CCl4 olive oil solution in the same way to induce acute liver injury in mice.

[0028] The mice were modeled for 12 hours, during which they were fasted but not watered. The weight of the mice was recorded after 12 hours. The mice were pre-anesthetized with isoflurane, the eyeballs were removed, blood was collected and the mice were killed. After the blood was allowed to stand at 4 °C for 30 minutes, it was centrifuged at 3500 rpm / min and 4 °C for 10 minutes. The supernatant was transferred to a clean centrifuge tube using a pipette to obtain serum. At the same time, the mouse liver tissue was taken, washed with saline and the surface water was removed. Part of the liver tissue was accurately weighed to prepare 10% liver homogenate. Another part of the liver tissue was fixed in 4% paraformaldehyde for 24 hours for pathological sections, and the rest of the liver was stored at -80 °C.

[0029] 1. Analysis of mouse liver pathological tissue sections Mouse liver tissue was obtained, fixed with 4% paraformaldehyde for 24 hours, embedded in paraffin and sliced. H&E staining was used to observe the changes in liver tissue under a microscope.

[0030] 2. Determination of liver function level in mice According to the kit instructions, the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (AKP) in mouse serum were detected.

[0031] 3. Determination of Antioxidant Capacity in Mice Part of the liver tissue was accurately weighed and added with physiological saline to prepare 10% liver tissue homogenate. The activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and malondialdehyde (MDA) in the liver tissue of mice were determined using a kit.

[0032] 4. Statistical analysis The data were statistically analyzed using GraphPad Prism 8.0 software, and one-way ANOVA was used between multiple groups. The results were expressed as mean ± standard deviation (X ± SD). P When the difference was less than 0.05, the difference between the groups was statistically significant.

[0033] Effects of Cinnamomum camphora polysaccharide on mice with acute liver injury 1. Effects of Cinnamomum camphora polysaccharide on pathological changes in mouse liver tissue Figure 1 The effects of Cinnamomum camphora polysaccharide on mouse liver tissue morphology are given. BC is the blank group, MC is the model group, PC is the bifendate positive control group, HD, MD, and LD are the high, medium, and low dose groups of Cinnamomum camphora polysaccharide, respectively. Figure 1 As shown in the figure, the liver tissue sections of the blank group were normal, the cell nuclei and nucleoli were clear, and the central vein was visible. In addition, the hepatic sinusoids were normal, and the hepatic plates and hepatic sinusoids were arranged regularly. In contrast, the hepatocytes in the model group were enlarged and irregularly arranged, the hepatic plates were arranged in disorder, the cell nuclei were atrophied, and the hepatocytes were necrotic in a large area with vacuolation. After treatment with tea fruit camphor polysaccharide, the liver lesions were significantly improved, the liver tissue cell structure was normal, and the structure was close to that of normal liver. The results showed that tea fruit camphor polysaccharide had a significant protective effect on CCl4-induced liver damage in mice.

[0034] 2. Effects of Cinnamomum camphora polysaccharide on liver function in mice Figure 2 The effects of tea fruit camphor polysaccharide on the activities of related enzymes in mouse serum are given. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (AKP) are important markers for clinical evaluation of liver damage, such as Figure 2 As shown in Figure 2, the levels of AST, ALT and AKP in the serum of mice in the model group were significantly higher than those in the blank control group ( P <0.001), indicating that CCl4 induced liver damage in mice. After drug treatment, the levels of PC group and drug treatment group were improved to varying degrees. These results show that the levels of AST, ALT and AKP in mice with CCl4-induced liver damage can be improved by Cinnamomum camphora polysaccharide, and the Cinnamomum camphora polysaccharide administration group showed a good dose-dependency in the reduction of liver function indicators in mice.

[0035] 3. Effect of Cinnamomum camphora polysaccharide on the antioxidant level of mice Figure 3 The effects of Cinnamomum camphora polysaccharide on superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-Px) activity and malondialdehyde (MDA) content in mouse liver were given. Figure 3 As shown in Figure 2, compared with the blank group, the activities of SOD and GSH-Px in the serum of mice in the model group were significantly reduced ( P<0.05), MDA content was significantly increased ( P <0.001). The drug group and PC group could reverse this decrease to varying degrees and showed a dose-dependent manner. At the same time, compared with the model group, the MDA content in the HD group and MD group was significantly reduced. The results showed that tea fruit camphor polysaccharide may protect the liver by enhancing the antioxidant capacity of mice.

[0036] Example 2 Preparation of tea fruit camphor polysaccharide: A method for preparing tea fruit cinnamon polysaccharide comprises the following steps: 254 g of Cinnamomum camphora fruit was crushed and extracted with petroleum ether and ethyl acetate at room temperature according to a solid-liquid ratio of 1g:20 ml, respectively. Each extraction was 12 h, and a total of 3 extractions were performed. After the extraction, the supernatant was filtered out and the residue was retained. The residue was heated and extracted at 80°C with water at a solid-liquid ratio of 1g:30 ml. The heating extraction time was 4 h each time, and the extraction was performed twice. After the water extracts were combined, they were concentrated under reduced pressure at 60°C using a rotary evaporator. 95% ethanol was added to the concentrate until the final ethanol concentration was 75%. The concentrate was allowed to stand for 24 hours, centrifuged, and the precipitate was collected. The precipitate was dissolved in water and then the protein was removed by the Sevage method (conventional techniques in the art can be used, or as follows: sample: Sevage reagent = 15:5, Sevage reagent is a chloroform-n-butanol mixture with a volume ratio of 4:1; oscillation for 60 minutes, standing for 30 minutes, centrifuged at 7000 r / min for 6 minutes, and the denatured protein outside the interface between the water layer and the chloroform layer was removed. The upper sugar solution was taken and the denatured protein removal operation was repeated until all the protein was removed). The supernatant was added with 95% ethanol until the final ethanol concentration was 75%, allowed to stand for 24 hours, centrifuged, and the precipitate was collected and freeze-dried (-50°C, 12 hours) to obtain 18.158 g of tea fruit cinnamomum camphora polysaccharide, with a yield of about 7.15% and a sugar content of 49.86%, which was set aside.

[0037] The protective effect of the cinnamomum camphora polysaccharide prepared in Example 2 on cyclophosphamide-induced oxidative stress mice was investigated below.

[0038] Application experiment 2 Cyclophosphamide-induced oxidative stress in mice: The tea fruit camphor polysaccharide prepared in Example 1 was subjected to in vivo animal experiments. 1. Experimental Animals Same as application experiment 1.

[0039] 2. Test reagents and instruments Electronic balance (ME204, METTLER TOLEDO) Rotary evaporator (N-1300, EYELA); FMB40 fully automatic flake ice machine (Shanghai Bilang Instrument Co., Ltd.); Desktop centrifuge, MiniSpin, Shanghai Anting Scientific Instrument Factory; Full wavelength microplate reader, 1510 Thermo Fisher; KZ-Ⅲ-FP high-speed cryogenic tissue grinder (Wuhan Saiweier Biotechnology Co., Ltd.); Electric heating blast drying oven (Shanghai Yiheng Scientific Instrument Co., Ltd.); Cyclophosphamide for injection (Jiangsu Hengrui Medicine Co., Ltd.); Lentinus edodes polysaccharide tablets (Hubei Guangren Pharmaceutical Co., Ltd.); Malondialdehyde (MDA) assay kit (Nanjing Jiancheng Bioengineering Institute); Catalase (CAT) assay kit (Nanjing Jiancheng Bioengineering Institute); Total antioxidant capacity (T-AOC) assay kit (Nanjing Jiancheng Bioengineering Institute); BCA protein concentration determination kit (Beijing Solebow Technology Co., Ltd.).

[0040] 3. Test methods Table 2 Experimental groups and corresponding dosages After 1 week of adaptive feeding, the mice were randomly divided into 6 groups: blank group, model group, positive drug group, low-dose group, medium-dose group, and high-dose group, with 8 mice in each group. The corresponding dosages are shown in Table 2. Each group of mice was gavaged once a day at a volume of 10 ml / kg for 3 weeks. All drugs were dissolved in physiological saline for administration.

[0041] The mice were weighed every 2 days, and the mice were gavaged daily, and the gavage volume was adjusted according to the weight changes of the mice. The blank group and the model group were given normal saline, the PC group was given Lentinan polysaccharide, and the treatment group was given Cinnamomum camphora polysaccharide. On the 18th, 19th, 20th, and 21st days of administration, the mice in the BC blank group were intraperitoneally injected with normal saline, and the mice in the other groups were intraperitoneally injected with cyclophosphamide CTX at a dose of 70 mg / kg·BW for modeling.

[0042] The mice were modeled for 12 hours, during which they were fasted but not watered. The weight of the mice was recorded after 12 hours. After pre-anesthesia with isoflurane, the eyeballs were removed to collect blood and the mice were killed. After the blood was allowed to stand at 4°C for 30 minutes, it was centrifuged at 3500 rpm / min and 4°C for 10 minutes. The supernatant was transferred to a clean centrifuge tube using a pipette to obtain serum. At the same time, the mouse liver tissue was taken, washed with saline and the surface water was removed. Part of the liver tissue was accurately weighed to prepare 10% liver homogenate. The rest of the liver was stored at -80°C.

[0043] 1. Determination of antioxidant capacity in mice Part of the liver tissue was accurately weighed and added with physiological saline to prepare 10% liver tissue homogenate. The contents of malondialdehyde (MDA), catalase (CAT), and total antioxidant capacity (T-AOC) in mouse liver tissue were determined using a kit.

[0044] 2. Statistical analysis The data were statistically analyzed using GraphPad Prism 8.0 software, and one-way ANOVA was used between multiple groups. The results were expressed as mean ± standard deviation (X ± SD). P When the difference was less than 0.05, the difference between the groups was statistically significant.

[0045] Effects of Cinnamomum camphora polysaccharide on cyclophosphamide-induced oxidative stress in mice 1. Effect of Cinnamomum camphora polysaccharide on the antioxidant level of mice Cyclophosphamide (CTX) is an alkylating cytotoxic drug commonly used to treat cancer, however, its high dose may cause immunosuppression and oxidative stress damage. Oxidative stress response leads to an imbalance between pro-oxidant and antioxidant factors, resulting in damage to cellular DNA, proteins, and lipids. The antioxidant capacity of mice can be assessed by detecting the malondialdehyde (MDA) content, catalase (CAT), and total antioxidant capacity (T-AOC) in liver tissue.

[0046] Figure 4 The results of the effects of tea fruit camphor polysaccharide on CAT (A) activity, T-AOC (B) activity and MDA (C) in mouse liver are given. Figure 4 As shown, compared with the blank group, the activity of catalase (CAT) (Figure A) and total antioxidant capacity (T-AOC) (Figure B) in the liver of mice in the model group was significantly reduced ( P <0.001), and the malondialdehyde (MDA) (Figure C) content was significantly increased. The high-dose group and PC group of the present invention can significantly reverse this decrease. At the same time, compared with the model group, the high-dose MDA content was significantly reduced. The results show that the present invention of the tea fruit camphor polysaccharide may achieve the effect of resisting oxidative stress caused by cyclophosphamide by enhancing the antioxidant capacity of mouse liver.

[0047] In summary, by using carbon tetrachloride to establish a mouse acute liver injury model, the present invention found that tea fruit camphor polysaccharide can effectively reduce the activity of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase in the serum of mice with acute liver injury, improve liver tissue morphology, increase the activity of superoxide dismutase (SOD) and glutathione catalase (GSH-Px) in mouse liver, reduce the content of malondialdehyde (MDA), and alleviate the damage of mouse liver tissue. It has excellent liver protection effect and has the potential to be developed into a liver protection drug, especially for preparing drugs that protect acute liver injury. In addition, by using cyclophosphamide to establish a mouse oxidative stress model, the present invention found that tea fruit camphor polysaccharide can also effectively reduce the content of malondialdehyde (MDA) in mouse liver, increase the level of catalase (CAT) and total antioxidant capacity (T-AOC) in mouse liver, have a good protective effect on mouse oxidative stress injury, have excellent anti-oxidative stress effect, and have the potential to be developed into an anti-oxidative stress drug.

Claims

1. A method for extracting tea fruit cinnamon polysaccharides, characterized in that: The steps include: The cinnamomum camphora fruit is crushed and then heated with water at 65-90°C for extraction, the water extract is concentrated under reduced pressure, the concentrate is precipitated with alcohol, protein is removed, and then precipitated with alcohol again, and freeze-dried to obtain cinnamomum camphora polysaccharide A; Alternatively, after crushing the fruits of Cinnamomum camphora, extracting them with petroleum ether and ethyl acetate at room temperature, separating the solid and liquid, extracting the residue with water at 65-90°C, concentrating the water extract under reduced pressure, precipitating the concentrate with alcohol, removing protein, precipitating again with alcohol, and freeze-drying to obtain Cinnamomum camphora polysaccharide B; The tea fruit cinnamon polysaccharide A and / or tea fruit cinnamon polysaccharide B are tea fruit cinnamon polysaccharides.

2. The method for extracting tea fruit cinnamon polysaccharides according to claim 1, characterized in that: Add water according to the solid-liquid ratio of 1g:20-40 ml for heating extraction. The heating extraction time is 2-5 hours, and the heating extraction is performed 1-4 times.

3. The method for extracting tea fruit cinnamon polysaccharides according to claim 1, characterized in that: Add petroleum ether or ethyl acetate according to the solid-liquid ratio of 1g:10-30 ml for extraction. The extraction time is 8-15 hours and the extraction is performed 1-5 times.

4. Cinnamomum camphora polysaccharide obtained by extraction using the extraction method according to any one of claims 1 to 3.

5. A compound preparation comprising the tea fruit cinnamon polysaccharide according to claim 4, characterized in that: The tea fruit camphor polysaccharide is compounded with conventional auxiliary materials in the art to prepare a compound preparation.

6. The compound preparation according to claim 5, characterized in that: The dosage form of the compound preparation is tablet, granule, pill, capsule or injection.

7. Use of the cinnamomum camphora polysaccharide according to claim 4 or the compound preparation according to claim 5 in the preparation of a drug for treating or preventing oxidative stress.

8. The use according to claim 7, characterized in that The oxidative stress is oxidative stress caused by cyclophosphamide; the Cinnamomum camphora polysaccharide or compound preparation can effectively reduce the malondialdehyde content in the liver and increase the levels of catalase and total antioxidant capacity in the liver.

9. Use of the cinnamomum camphora polysaccharide according to claim 4 or the compound preparation according to claim 5 in the preparation of a drug for treating or preventing liver damage.

10. The use according to claim 9, characterized in that The liver damage is acute liver damage; the camphor tree polysaccharide or compound preparation can reduce the levels of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase, and alleviate the damage to the liver organ.

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