Tibetan medicine for relieving liver injury caused by potassium dichromate as well as preparation method and application of Tibetan medicine

Abstract

The invention discloses a preparation method and application of a Tibetan medicine capable of relieving liver injury caused by potassium dichromate, and belongs to the technical field of Tibetan medicines.The preparation method of the Tibetan medicine specifically comprises the following steps that 1, trametes versicolor sporocarp is taken and ground into coarse powder, the coarse powder is added into a 95% ethanol solution, after ultrasonic treatment is conducted for 30 min, filtering is conducted, and a mixture is obtained; (2) adding sterile water into the obtained mixture for extraction, and repeating the process twice to obtain an extract; (3) performing deproteinization treatment on the obtained extract by using a Sevag reagent to obtain a crude polysaccharide solution; and (4) pouring the obtained crude polysaccharide solution into a dialysis bag with the molecular weight cutoff value of 3500Da, collecting the polysaccharide solution in the dialysis bag, and freeze-drying to obtain coriolus versicolor polysaccharide powder. The coriolus versicolor polysaccharide disclosed by the invention not only can be used for improving the antioxidant capacity of mice and relieving liver injury, but also can be used for promoting the recovery of body functions by improving intestinal flora composition and regulating metabolic pathways.

Description

Technical Field

[0001] The present invention belongs to the technical field of Tibetan medicine, and particularly relates to a Tibetan medicine for relieving liver injury caused by potassium dichromate, a preparation method thereof, and an application thereof. Background Art

[0002] Liver injury refers to the structural and functional damage that occurs when the liver is affected by various factors (such as drugs, poisons, viral infections, alcoholism, etc.). As a crucial metabolic organ in the body, the liver undertakes many important functions such as detoxification, protein synthesis, and energy storage. Once the liver is damaged, serious health problems may be triggered. Current treatment methods usually rely on antibiotics. However, with the long-term and inappropriate use of antibiotics, it is easy to cause an increase in drug residues and drug resistance. At the same time, it will also cause drug residues in the body, produce toxic and side effects on the body, and due to incomplete treatment, it is easy to relapse, and may even further aggravate liver injury.

[0003] Coriolus versicolor, also known as Trametes versicolor or Coriolus versicolor, is a type of Coriolus versicolor that grows in high-altitude areas such as the Qinghai-Tibet Plateau. It belongs to the family Polyporaceae and is a widely recognized traditional Chinese medicine. Coriolus versicolor is rich in various active ingredients due to its unique growth environment and has high medicinal value and health care effects. In traditional Chinese medicine, Coriolus versicolor is often used in aspects such as enhancing the immune system, antioxidation, anti-tumor, and regulating body functions. It contains a variety of complex chemical components, including fats, polysaccharide peptides, amino acids, proteins, inorganic salts, alkaloids, and various enzyme substances. These components work together to endow Coriolus versicolor with various pharmacological activities. In particular, the polysaccharide extracted from Coriolus versicolor, which is a high-molecular polymer, has attracted much attention due to its significant effects in immunomodulation, anti-cancer and anti-tumor, anti-inflammatory, improving immunity, lowering blood sugar and blood lipids, antioxidation, protecting the liver, and anti-diarrhea, and the polysaccharide of Coriolus versicolor does not produce side effects such as drug resistance.

[0004] Therefore, in view of the above characteristics, Coriolus versicolor and its main active ingredient, polysaccharide of Coriolus versicolor, as an efficient, safe and drug that is not prone to drug resistance, provide new possibilities for relieving liver injury and have become one of the directions that need to be explored and studied by those skilled in the art. Summary of the Invention

[0005] In view of the deficiencies of the above-mentioned prior art, the present invention discloses a Tibetan medicine for relieving liver injury caused by potassium dichromate, a preparation method thereof, and an application thereof. The Tibetan medicine disclosed by the present invention is applied in mouse experiments and can effectively relieve liver injury in mice, enhance the overall physical fitness of mice, improve their antioxidant capacity, regulate the composition and structure of the intestinal flora of mice, enhance the intestinal barrier function, and enhance the immune system efficacy, so as to achieve the effects of relieving liver injury in mice, restoring their body functions, and enhancing immunity.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention provides a preparation method of a Tibetan medicine for relieving liver injury caused by potassium dichromate, which is characterized by comprising the following steps:

[0008] (1) Take an appropriate amount of dried Ganoderma tsugae fruiting bodies, grind them into coarse powder, add the Ganoderma tsugae coarse powder to a 95% ethanol solution, perform ultrasonic treatment in an ultrasonic cleaner for 30 min, and then filter to obtain a mixture;

[0009] (2) Add sterile water to the mixture obtained in step (1) for extraction, repeat this process twice, combine the aqueous extracts obtained from the two extractions to obtain an extract;

[0010] (3) Use Sevag reagent (a chloroform-n-butanol mixed solvent) to perform deproteinization treatment on the extract obtained in step (2), fully mix the solution with the Sevag reagent and then let it stand for stratification, remove the protein layer, and retain the aqueous phase containing crude polysaccharides to obtain a solution of crude polysaccharides;

[0011] (4) Pour the crude polysaccharide solution obtained in step (3) into a dialysis bag with a molecular weight cut-off value of 3500 Da, place it in a large amount of sterile water for dialysis for 48 h, after dialysis, collect the polysaccharide solution in the dialysis bag, and perform freeze-drying using a freeze dryer to obtain Ganoderma tsugae polysaccharide powder, that is: a Tibetan medicine capable of relieving liver injury caused by potassium dichromate is prepared.

[0012] Preferably, in step (1), the mass-volume ratio of the Ganoderma tsugae coarse powder to the 95% ethanol solution is 1 g:(15 - 30) mL.

[0013] Preferably, in step (2), the mass-volume ratio of the mixture to sterile water is 1 g:(8 - 16) mL; the extraction temperature is 100 °C and the extraction time is 90 min.

[0014] Preferably, in step (3), the volume ratio of chloroform to n-butanol in the Sevag reagent is 4:1.

[0015] Preferably, in step (4), the temperature of the freeze-drying is -80 °C and the freeze-drying time is 48 h.

[0016] The present invention discloses a Tibetan medicine prepared by the above preparation method and capable of relieving liver injury caused by potassium dichromate.

[0017] The present invention also discloses an application of the above Tibetan medicine in the preparation of a drug product having the effect of relieving liver injury.

[0018] Preferably, the drug product for relieving liver injury is a drug product for relieving or treating liver injury caused by potassium dichromate.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] After the application of Coriolus versicolor polysaccharide provided by the present invention to mice, it is found through experiments that: (1) Coriolus versicolor polysaccharide can significantly improve the antioxidant capacity of mice, help enhance the body's ability to resist oxidative stress, and reduce the damage of free radicals to cells (such as Figure 2 ). At the same time, Coriolus versicolor polysaccharide can effectively relieve liver injury caused by potassium dichromate, has the potential to protect the liver from chemical damage, and provides new ideas and methods for the treatment of liver diseases ( Figure 3 ). In addition, regarding the influence of Coriolus versicolor polysaccharide on the intestinal flora of mice, it improves intestinal performance by enriching beneficial bacteria and reducing harmful bacteria, and regulates the composition and structure of the intestinal flora. This adjustment not only promotes the healthy changes of the intestinal flora of mice, but also improves intestinal immunity and further enhances the overall health status of the body (such as Figure 7 ). Moreover, Coriolus versicolor polysaccharide can regulate the body's functions by affecting the abundance of specific metabolites and their related metabolic pathways. For example, Coriolus versicolor polysaccharide can regulate important pathways such as starch and sucrose metabolism, and nucleotide sugar biosynthesis, help restore the normal physiological state of mice, and relieve liver injury caused by potassium dichromate (such as Figure 13 ).

[0021] In summary, the application of Coriolus versicolor polysaccharide can not only improve the antioxidant capacity of mice, relieve liver injury, but also promote the recovery of the body's functions by improving the composition of the intestinal flora and regulating metabolic pathways. This provides important scientific basis and new treatment references for exploring the application of Coriolus versicolor polysaccharide in the treatment of liver injury and other related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is an analysis diagram of the body state of mice in the present invention (a is the body weight of mice, b is the net weight rate, c is the organ weight);

[0023] Figure 2 It is an analysis diagram of the serum indexes of mice in the present invention (a is liver function, b is kidney function, c is inflammatory factor index, d is oxidative stress ability);

[0024] Figure 3 It is an analysis diagram of HE and Sirius red stained sections of the liver and kidneys of mice in the present invention (a is HE staining, b is Sirius red staining);

[0025] Figure 4 It is the result of analyzing the intestinal gene expression level of mice by RT-qPCR (real-time quantitative polymerase chain reaction) in the present invention;

[0026] Figure 5 Analysis results of the protein expression level in the liver tissue of mice in the present invention;

[0027] Figure 6 Analysis results of the Venn diagram and α-diversity of the intestinal flora of mice in the present invention (a is the Venn diagram, b is the rarefaction curve, c is the rank-abundance curve, d is the α-diversity index);

[0028] Figure 7 Analysis results of the microbial community structure of mice at different taxonomic levels in the present invention (a is the phylum level, b is the class level, c is the order level, d is the family level, e is the genus level);

[0029] Figure 8 Analysis results of the β-diversity and heat map analysis of the intestinal flora of mice in the present invention (a is the β-diversity, b is the heat map);

[0030] Figure 9 Analysis results of the LEfSe analysis of the intestinal flora of mice in the present invention;

[0031] Figure 10 Analysis of the marker species in different groups of mice by T-test in the present invention (a is the phylum level, b is the genus level);

[0032] Figure 11 Comparison results of the functions of the microbiota of mice in different groups in the present invention (a is KEGG, b is MetaCyc);

[0033] Figure 12 Comparison analysis chart of the metabolites of mice in different groups in the present invention (a is group comparison, b is Venn diagram);

[0034] Figure 13 Comparison analysis results of the KEGG pathways of the metabolites of mice in different groups in the present invention. Detailed implementation manners

[0035] 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 the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0036] The experimental methods used in the following embodiments are all conventional methods unless otherwise specified.

[0037] The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0038] The present invention provides a preparation method of a Tibetan medicine for alleviating liver injury caused by potassium dichromate, comprising the following steps:

[0039] (1) Take an appropriate amount of dried Ganoderma tsugae fruiting bodies, grind them into coarse powder, mix the Ganoderma tsugae coarse powder with 95% ethanol solution at a mass-to-volume ratio of 1 g:(15 - 30) mL, and after ultrasonic treatment in an ultrasonic cleaner for 30 min, filter to obtain a mixture.

[0040] (2) Mix the mixture obtained in step (1) with sterile water at a mass-to-volume ratio of 1 g:(8 - 16) mL, and perform extraction under the conditions of an extraction temperature of 100 °C and an extraction time of 90 min. Repeat this process twice, and combine the water extracts obtained from the two extractions to obtain an extract.

[0041] (3) Use Sevag reagent (wherein: the volume ratio of chloroform to n-butanol is 4:1) to perform protein removal treatment on the extract obtained in step (2). After fully mixing the solution with Sevag reagent, let it stand for stratification, remove the protein layer, and retain the aqueous phase containing crude polysaccharides to obtain a solution of crude polysaccharides.

[0042] (4) Pour the crude polysaccharide solution obtained in step (3) into a dialysis bag with a molecular weight cut-off value of 3500 Da, place it in a large amount of sterile water for dialysis for 48 h. After dialysis, collect the polysaccharide solution inside the dialysis bag. Place the polysaccharide solution collected inside the above dialysis bag in a freeze dryer and freeze-dry it for 48 h at a temperature of -80 °C to obtain Ganoderma tsugae polysaccharide powder, that is: a Tibetan medicine capable of alleviating liver injury caused by potassium dichromate is prepared.

[0043] The present invention discloses a Tibetan medicine prepared by the above preparation method and capable of alleviating liver injury caused by potassium dichromate.

[0044] The present invention also discloses an application of the above Tibetan medicine in the preparation of a pharmaceutical product having the effect of alleviating liver injury.

[0045] Among them, the pharmaceutical product having the effect of alleviating liver injury may be a pharmaceutical product for alleviating or treating liver injury caused by potassium dichromate.

[0046] The technical solution of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments:

[0047] Example 1

[0048] In this example, the preparation method of the Tibetan medicine for alleviating liver injury caused by potassium dichromate is as follows:

[0049] (1) Take an appropriate amount of dry Coriolus versicolor fruiting bodies, grind them into coarse powder, mix the Coriolus versicolor coarse powder with 95% ethanol solution at a mass-to-volume ratio of 1 g:20 mL, and perform ultrasonic treatment in an ultrasonic cleaner for 30 min, then filter to obtain a mixture.

[0050] (2) Mix the mixture obtained in step (1) with sterile water at a mass-to-volume ratio of 1 g:10 mL, and perform extraction under the conditions of an extraction temperature of 100 °C and an extraction time of 90 min. Repeat this process twice, and combine the aqueous extracts obtained from the two extractions to obtain an extract.

[0051] (3) Use Sevag reagent (wherein the volume ratio of chloroform to n-butanol is 4:1) to perform deproteinization treatment on the extract obtained in step (2). After the solution is fully mixed with the Sevag reagent, let it stand and separate into layers, remove the protein layer, and retain the aqueous phase containing crude polysaccharide to obtain a solution of crude polysaccharide.

[0052] (4) Pour the crude polysaccharide solution obtained in step (3) into a dialysis bag with a molecular weight cut-off value of 3500 Da, place it in a large amount of sterile water for dialysis for 48 h. After dialysis, collect the polysaccharide solution inside the dialysis bag. Place the polysaccharide solution collected inside the above dialysis bag in a freeze dryer and freeze-dry it at a temperature of -80 °C for 48 h to obtain Coriolus versicolor polysaccharide powder, that is, a Tibetan medicine capable of alleviating liver injury caused by potassium dichromate is prepared.

[0053] Example 2

[0054] This example is an experiment on the effects of the Tibetan medicine (Coriolus versicolor polysaccharide powder) prepared in Example 1 above on the body weight, organs, and serum indicators of mice.

[0055] Thirty 4-week-old BALB / C mice with a body weight of 23.18 ± 1.26 g were selected and randomly divided into three groups of 10 mice each, namely the CCR group (blank control group), the CMR group (potassium dichromate modeling group), and the CYR group (polysaccharide from Coriolus versicolor treatment group). After 3 days of adaptive feeding, all mice started the formal experiment. The entire experimental period was 35 days. During the experiment, the mice in the CMR group and the CYR group were given intragastric administration of potassium dichromate (K2Cr2O7, dose: 10 mg / kg) every day. Meanwhile, the CYR group received intragastric administration of polysaccharide from Coriolus versicolor (dose: 25 mg / kg) for treatment, while the CCR group and the CMR group were given intragastric administration of an equal amount of sterile water as a control. All mice were housed under a 12-hour light / dark cycle and had free access to food and water. At the end of the experiment on the 36th day, the body weight of each mouse was recorded, and blood samples were collected by orbital blood sampling. After the blood samples were left standing at room temperature for 2 - 3 hours, the serum was separated by centrifugation at 3500 r / min for 15 min and used to detect the antioxidant capacity and immune capacity indexes of the mice, including total antioxidant capacity (T-AOC), superoxide dismutase (SOD), malondialdehyde (MDA), interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), as well as liver and kidney function biomarkers such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, and blood urea nitrogen (BUN).

[0056] Subsequently, all mice were euthanized by inhaling carbon dioxide, and the weights of the heart, liver, spleen, lungs, and kidneys were measured. Some liver and kidney tissues were preserved in 4% paraformaldehyde for subsequent HE staining and Sirius red staining analysis, while other liver tissues were used for protein extraction for RT-qPCR and Western Blot assays to comprehensively evaluate the therapeutic effect of the Tibetan medicine (polysaccharide from Coriolus versicolor) on mice with liver injury induced by potassium dichromate. The specific test results are as follows.

[0057] (1) As Figure 1 shown, (i) As can be seen from Figure 1 a: The body weight of the mice in the CCR group continued to increase steadily throughout the experimental period, showing a normal growth trend. Compared with the CCR group, the body weight of the mice in the CMR group was significantly lower than that in the CCR group during the experiment, indicating that potassium dichromate (K2Cr2O7) had a negative impact on the growth of the mice. The body weight of the mice in the CYR group treated with polysaccharide from Coriolus versicolor increased significantly, and its growth trend was close to that of the CCR group, indicating that polysaccharide from Coriolus versicolor could effectively promote the growth of the mice and relieve the weight loss caused by potassium dichromate. (ii) From Figure 1It can be seen from Figure b that the net weight rate of the CCR group is relatively high, indicating a normal growth state; the net weight rate of the CMR group is significantly decreased (p < 0.05), further confirming the inhibitory effect of potassium dichromate on the growth of mice; after the mice in the CYR group were treated with Coriolus versicolor polysaccharide, the net weight rate of the mice was significantly increased, approaching the level of the CCR group, indicating that Coriolus versicolor polysaccharide has the effect of promoting the growth of mice. (ⅲ) In the CMR group, the heart index (p < 0.01) and kidney index (p < 0.0001) of the mice in the CMR group were significantly decreased, but the liver index (p < 0.001) and spleen index (p < 0.05) were significantly increased; however, after the mice in the CYR group were treated with Coriolus versicolor polysaccharide, the heart index (p < 0.05) and kidney index (p < 0.0001) of the mice were increased, while the liver index (p = 0.0001) and spleen index (p < 0.05) were decreased. This shows that potassium dichromate (K2Cr2O7) has obvious negative effects on the growth and organ function of mice, and the Coriolus versicolor polysaccharide of the present invention can effectively alleviate these adverse effects, promote the growth of mice, and improve the functions of the heart, kidney, liver and spleen.

[0058] (2) As Figure 2 shown, (ⅰ) From Figure 2 a, Figure 2 b, it can be seen that the contents of AST (p < 0.0001), ALT (p < 0.0011), Bun (p < 0.001) and creatinine (p < 0.00001) in the mice of the CMR group were significantly increased, while after the mice in the CYR group were treated with Coriolus versicolor polysaccharide, the contents of AST (p = 0.0001), ALT, Bun and creatinine (p < 0.001) in the mice were significantly decreased. This shows that potassium dichromate (K2Cr2O7) has significant negative effects on the liver function and kidney function of mice, resulting in obvious increases in the contents of AST, ALT, Bun and creatinine, while Coriolus versicolor polysaccharide can significantly reduce these indexes and restore the liver and kidney functions of mice. (ⅱ) From Figure 2It can be seen from Figure c that the levels of pro-inflammatory factors such as interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the CCR group were relatively low, while the level of the anti-inflammatory factor interleukin-10 (IL-10) was relatively high, indicating a normal inflammatory state; in the CMR group, IL-6 (p < 0.001), IL-1β (p < 0.001), and TNF-α (p < 0.01) were significantly increased, while IL-10 (p < 0.05) was significantly decreased, suggesting that potassium dichromate induced a strong inflammatory response, leading to an increase in the levels of pro-inflammatory factors and a decrease in the levels of anti-inflammatory factors; after treatment with Coriolus versicolor polysaccharide in the CYR group, IL-6 (p < 0.05), IL-1β (p < 0.05), and TNF-α (p < 0.001) were significantly decreased, while IL-10 (p < 0.01) was significantly increased, indicating that Coriolus versicolor polysaccharide can regulate the inflammatory response, reduce the levels of pro-inflammatory factors, and increase the levels of anti-inflammatory factors. (ⅲ) From Figure 2 It can be seen from Figure d that in the CCR group: the total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA) contents of the mice were all within the normal range; in the CMR group, the T-AOC (P < 0.05), GSH-Px (P < 0.01), and SOD (P < 0.05) of the mice in the CMR group were significantly decreased, while the MDA content was significantly increased (p < 0.0001), indicating that potassium dichromate induced lipid peroxidation in the body, generating a large number of free radicals; after treatment with Coriolus versicolor polysaccharide in the CYR group, the T-AOC (p < 0.001), GSH-Px (p < 0.01), and SOD (p < 0.01) were significantly increased, while the MDA content was significantly decreased (p < 0.01), indicating that Coriolus versicolor polysaccharide can enhance the antioxidant capacity of the mice, reduce the generation of free radicals, and protect the body from oxidative damage. It can be seen from this that potassium dichromate (K2Cr2O7) had a significant negative impact on the antioxidant capacity and inflammatory response of the mice, resulting in a decrease in the levels of T-AOC, GSH-Px, and SOD, an increase in the MDA level, as well as an increase in the levels of pro-inflammatory factors and a decrease in the levels of anti-inflammatory factors, while Coriolus versicolor polysaccharide could significantly improve these indicators, enhance the antioxidant capacity, and inhibit oxidative stress and inflammatory responses.

[0059] (3) As Figure 3 shown, the HE staining results ( Figure 3 a1 and Figure 3 a2): (ⅰ) From Figure 3As can be seen from a1, the liver cell structure in the CCR group was normal, without obvious damage. The hepatic lobule structure was clear, the hepatocytes were arranged neatly, and there was no inflammatory cell infiltration or bleeding. In the CMR group, obvious degeneration, inflammatory cell infiltration and bleeding of liver cells were observed. Hepatocyte swelling and necrosis could be seen, inflammatory cells aggregated in the damaged area, and there were local bleeding points. In the CYR group, after treatment with Coriolus versicolor polysaccharide, the liver injury was significantly reduced. Although there were still some mild cell degeneration and inflammatory cell infiltration, these injuries were significantly reduced compared with the CMR group, and the hepatocyte structure tended to be normal. (ii) As can be seen from Figure 3 a2, the renal tissue structure in the CCR group was normal, the renal tubules and glomeruli had clear morphology, and no obvious damage was seen. In the CMR group, damage to the renal tissue occurred, manifested as degeneration and exfoliation of renal tubular epithelial cells and interstitial inflammatory reaction. Protein casts could be seen in the renal tubular lumen, indicating impaired renal function. In the CYR group, after treatment with Coriolus versicolor polysaccharide, the renal injury was improved, the structures of the renal tubules and glomeruli tended to be normal, the inflammatory cell infiltration decreased, and the renal function recovered somewhat.

[0060] Results of Sirius red staining (Figs. b1 and b2): (i) As can be seen from Figure 3 b1, the degree of liver tissue fibrosis in the CCR group was low, the collagen fibers were evenly distributed, and no obvious fibrosis was seen. In the CMR group, liver tissue fibrosis was obvious, the collagen fibers increased, and fibrous septa were formed, resulting in disordered liver structure. In the CYR group, after treatment with Coriolus versicolor polysaccharide, the degree of liver fibrosis was significantly reduced, the collagen fiber content decreased, the fibrous septa became narrower, and the liver structure tended to be normal. (ii) As can be seen from Figure 3 b2, the degree of renal tissue fibrosis in the CCR group was low, and the collagen fibers were normally distributed around the renal tubules and glomeruli. In the CMR group, renal tissue fibrosis increased, the collagen fibers increased around the renal tubules and in the interstitial area, affecting renal function. In the CYR group, after treatment with Coriolus versicolor polysaccharide, the degree of renal fibrosis was reduced, the collagen fiber content decreased, and the structures of the renal tubules and glomeruli tended to be normal.

[0061] From the results of the above HE staining and Sirius red staining, it can be seen that obvious damage and fibrosis occurred in the liver and kidney tissues of the mice in the CMR group. After treatment with Coriolus versicolor polysaccharide, the degree of damage and fibrosis in the liver and kidney tissues of the mice in the CYR group was significantly reduced, indicating that Coriolus versicolor polysaccharide has the effect of protecting the liver and kidney from the damage and fibrosis caused by potassium dichromate.

[0062] (4) As Figure 4As shown, it can be seen from the figure that: In the CMR group of mice, the expressions of NQO-1 (P<0.001), Nrf-2 (P<0.01), and IL-10 (P<0.001) were inhibited, while the expressions of IL-6 (P<0.0001), TNF-α (P<0.001), and Bax (P<0.001) were promoted. In the CYR group, the expression levels of NQO-1 (P<0.01), Nrf-2 (P<0.01), and IL-10 (P<0.05) were relatively high in the mice treated with Coriolus versicolor polysaccharide; the levels of IL-6 (P<0.0001), TNF-α (P<0.01), and Bax (P<0.001) were also relatively high, but compared with the CMR group, their expression levels were relatively lower. This may be because Coriolus versicolor polysaccharide partially counteracts the effects of pro-inflammatory and pro-apoptotic genes by upregulating antioxidant and anti-inflammatory genes (such as NQO-1, Nrf-2, and IL-10). This indicates that in the CMR group of mice, due to the treatment with potassium dichromate, the expressions of antioxidant and anti-inflammatory genes in the body decreased, while the expressions of pro-inflammatory and pro-apoptotic genes increased, thus triggering oxidative stress and inflammatory responses; after treatment with Coriolus versicolor polysaccharide, the expressions of antioxidant and anti-inflammatory genes in the CYR group of mice were significantly upregulated, partially counteracting the effects of pro-inflammatory and pro-apoptotic genes, thereby reducing oxidative stress and inflammatory responses and protecting the body from damage.

[0063] (5) As Figure 5 shown, it can be seen from the figure that: The expression level of Bax protein in the CMR group was significantly higher than that in the CCR group and the CYR group, while the expression level of Bax protein in the CYR group was significantly lower than that in the CMR group and was close to the level of the CCR group. This indicates that in the CMR group of mice, due to the treatment with potassium dichromate, the expression of Bax protein in the body increased significantly, thus triggering apoptosis; after treatment with Coriolus versicolor polysaccharide, the expression level of Bax protein in the CYR group of mice decreased significantly and was close to the normal level, indicating that Coriolus versicolor polysaccharide can effectively inhibit apoptosis and protect the body from damage caused by potassium dichromate.

[0064] Example 3

[0065] This example is an experiment on the effect of the Tibetan medicine (Coriolus versicolor polysaccharide powder) prepared in the above Example 1 on the intestinal flora of mice.

[0066] Thirty 4-week-old BALB / C mice with a body weight of 23.18 ± 1.26 g were selected and randomly divided into three groups of 10 mice each, namely the CCR group (blank control group), the CMR group (potassium dichromate modeling group), and the CYR group (polysaccharide from Coriolus versicolor treatment group). After 3 days of adaptive feeding, all mice started the formal experiment. The entire experimental period was 35 days. During the experiment, the mice in the CMR group and the CYR group were given intragastric administration of potassium dichromate (K2Cr2O7, dose of 10 mg / kg) every day, and at the same time, the mice in the CYR group received intragastric treatment with polysaccharide from Coriolus versicolor (dose of 25 mg / kg). The CCR group and the CMR group were given intragastric administration of an equal amount of sterile water as a control. All mice were housed under a 12-hour light / dark cycle and had free access to food and water.

[0067] At the end of the experiment on the 36th day, all the above mice were euthanized by inhaling carbon dioxide. Subsequently, fresh rectal fecal samples of each group of mice were collected for 16S rRNA high-throughput sequencing to analyze the specific effects of the polysaccharide from Coriolus versicolor of the present invention on the intestinal flora of mice. The specific test results are as follows.

[0068] (1) As Figure 6 shown, (i) As can be seen from Figure 6 a: There were 9356 ASVs in the CCR group, 14277 ASVs in the CMR group, and 9531 ASVs in the CYR group. There were 656 common ASVs among these three groups, indicating a certain commonality among the three groups. (ii) As can be seen from Figure 6 b, Figure 6 c of the rarefaction curve and rank-abundance curve: In the rarefaction curve ( Figure 6 b), the rarefaction curves of each group of mice tended to be flat, indicating that the obtained samples had high diversity and uniformity, and the sampling depth was sufficient to cover most microbial species; in the rank-abundance curve ( Figure 6 c), the rank-abundance curves of each group also showed a horizontal trend, further verifying the diversity and uniformity of the samples, indicating that the microbial distribution among different groups was relatively stable. (iii) As can be seen from Figure 6 d: The α-diversity analysis showed that the Faith_pd index of the CMR group was significantly higher than that of the CCR group and the CYR group (P < 0.05), indicating that the species diversity of the CMR group was higher, which might be related to the effect of potassium dichromate on the intestinal flora; other α-diversity indices such as Chao1, ACE, Shannon, and Simpson showed no significant differences among the three groups, indicating that except for species diversity, the diversity in other aspects remained relatively consistent among different groups.

[0069] (2) As Figure 7 shown, (i) As can be seen from Figure 7From the horizontal analysis of the middle door, it can be seen that the main phyla in the CCR group are Firmicutes_D (34.26%), Bacteroidota (31.00%), and Campylobacterota (19.85%), indicating that Firmicutes and Bacteroidota are dominant in the CCR group; the main phyla in the CMR group are Bacteroidota (44.00%), Firmicutes_A (17.96%), and Desulfobacterota_I (12.28%). Compared with the CCR group, the proportion of Bacteroidota in the CMR group increased significantly, while the proportion of Firmicutes was relatively low; the main phyla in the CYR group are Bacteroidota (37.99%), Firmicutes_D (33.42%), and Verrucomicrobiota (9.48%). The proportions of Bacteroidota and Firmicutes in the CYR group are close, and there is a certain proportion of Verrucomicrobiota. (ii) From Figure 7 From the class-level analysis in b, it can be seen that the main classes in the CCR group are Bacilli (34.26%), Bacteroidia (40.00%), and Campylobacteria (19.85%), indicating that Lactobacilli and Bacteroidia are dominant in the CCR group; the main classes in the CMR group are Bacteroidia (44.00%), Clostridia_258483 (17.96%), and Desulfovibrionia (12.28%). The proportion of Bacteroidia in the CMR group increased further, and a certain proportion of Desulfovibrionia appeared; CYR group: the main classes are Bacteroidia (37.99%), Bacilli (33.43%), and Verrucomicrobiae (9.48%). The proportions of Bacteroidia and Lactobacilli in the CYR group are close, and there is a certain proportion of Verrucomicrobiae. (iii) From Figure 7Horizontal analysis at the order level in c shows that the dominant taxa in the CCR group are Lactobacillales (32.47%), Bacteroidales (30.73%) and Campylobacterales (19.86%), indicating that Lactobacillales and Bacteroidales are dominant in the CCR group; the dominant taxa in the CMR group are Bacteroidales (42.67%), Desulfovibrionales (12.43%) and Lactobacillales (10.91%). The proportion of Bacteroidales in the CMR group increased significantly, and a certain proportion of Desulfovibrionales appeared at the same time; the dominant taxa in the CYR group are Bacteroidales (36.96%), Lactobacillales (32.94%) and Verrucomicrobiales (9.55%). The proportions of Bacteroidales and Lactobacillales in the CYR group are close, and there is a certain proportion of Verrucomicrobiales. (ⅳ) From Figure 7 Horizontal analysis at the family level in d shows that the main ones in the CCR group are Lactobacillaceae (31.93%), Muribaculaceae (24.64%) and Helicobacteraceae (19.89%), indicating that Lactobacillaceae and Muribaculaceae are dominant in the CCR group; the main ones in the CMR group are Muribaculaceae (30.23%), Desulfovibrionaceae (12.61%) and Lachnospiraceae (10.91%). The proportion of Muribaculaceae in the CMR group is relatively high, and a certain proportion of Desulfovibrionaceae and Lactobacillaceae appear at the same time; the main ones in the CYR group are Lactobacillaceae (33.03%), Muribaculaceae (26.51%) and Akkermansiaceae (9.90%). The proportions of Lactobacillaceae and Muribaculaceae in the CYR group are relatively high, and there is a certain proportion of Akkermansiaceae. (ⅴ) From Figure 7Horizontal analysis of the genera in e showed that the main genera in the CCR group were Lactobacillus (19.65%), Ligilactobacillus (13.57%), and Mucispirillum (10.63%), indicating that the genera Lactobacillus and Mucispirillum were dominant in the CCR group; the main genera in the CMR group were Mailhella (13.32%), Paramuribaculum (9.10%), and Lactobacillus (7.72%). The proportions of Mailhella and Paramuribaculum were relatively high in the CMR group, but the proportion of Lactobacillus was relatively low; the main genera in the CYR group were Lactobacillus (25.54%), Akkermansia (11.55%), and Paramuribaculum (10.14%). The proportions of Lactobacillus and Akkermansia were relatively high in the CYR group, and there was a certain proportion of Paramuribaculum.

[0070] From the above analysis, it can be seen that there were significant differences in the intestinal flora of mice at the phylum, class, order, family, and genus levels. Potassium dichromate treatment (CMR group) led to a significant increase in the proportions of Bacteroidetes and its related classes, orders, families, and genera, while polysaccharide treatment of Coriolus versicolor (CYR group) partially restored the flora structure, but there were still certain differences. The above detection and analysis results illustrate the regulatory effect of polysaccharide of Coriolus versicolor on the intestinal flora.

[0071] (3) As Figure 8 shown, (i) β-diversity analysis ( Figure 8 a1, Figure 8 a2, Figure 8 a3): From the PCoA analysis in Figure 8 a1, it can be seen that through PCoA (principal coordinates analysis), the relative distance between the CCR group and the CYR group was closer than that between the CCR group and the CMR group, indicating that in terms of β-diversity, the flora structure of the polysaccharide treatment group of Coriolus versicolor (CYR group) was more similar to that of the blank control group (CCR group); from the NMDS (non-metric multidimensional scaling analysis) in Figure 8 a2, this result was further verified. The positions of the CCR group and the CYR group were close in the NMDS plot, while the CMR group was relatively far away, indicating that potassium dichromate treatment had a significant impact on the intestinal flora structure; from the UPGMA (unweighted pair-group method with arithmetic mean) clustering tree in Figure 8 a3, a similar trend was also revealed. The CCR group and the CYR group clustered into one branch, while the CMR group clustered into another branch alone, once again proving the differences among the three groups; at the same time, the PERMANOVA (permutation-based multivariate analysis of variance) results showed that there were significant differences among the three groups (p < 0.05), further confirming the reliability of the above analysis. (ii) Heat map analysis ( Figure 8 b1, Figure 8b2): From Figure 8 It can be seen from the phylum-level heatmap of b1 that the proportions of Firmicutes_C, Firmicutes_G, and Campylobacterota are relatively high in the CCR group, the proportions of Proteobacteria, Firmicutes_B370539, Deinococcota, Patescibacterota, Plantomycetota, Cyanobacteria, Firmicutes_A, and Actinobacteriota are relatively high in the CMR group, and the proportions of Verruomicrota, Nitrospirota_A_437815, Eisenbacteria, Gemmatimonadota, Methylomirabilota, Myxococcota_A_473307, and Acidobacteriota are relatively high in the CYR group; From Figure 8 It can be seen from the genus-level heatmap of b2 that the proportions of Rikenelia, UBA7173, Mammaliicoccus_3179276, Corynebacterium, UBA3282, Psychrobacter, Facklamia_A_32262, Aerococcus, Clostridium_T, Enterenecus, Staphylococcus, COE1, CAG-485, Desulfovibrio_R_446353, Kineothrix, and Lawsonibacter are relatively high in the CMR group, the proportions of Helicobacter_D, Mucispirillum, Helicobacter_C_479931, Ligilactobacillus, Ruminococcus_C_58660, and Malacoplasma_A_271108 are relatively high in the CCR group, and the proportion of Eubacterium_R is relatively high in the CYR group.

[0072] The above β-diversity analysis shows that the polysaccharide of Coriolus versicolor treatment (CYR group) partially restored the changes in the intestinal flora structure caused by potassium dichromate treatment (CMR group), making it closer to the blank control group (CCR group). PERMANOVA analysis further confirmed the significant differences among the three groups. Heatmap analysis detailedly demonstrated the dominant flora among groups at the phylum and genus levels, revealing the regulatory effect of the polysaccharide of Coriolus versicolor on the intestinal flora.

[0073] (4) Such as Figure 9As shown, the LEfSe analysis showed that in the CCR group, Mycoplasmatales (p<0.05), Malacoplasma_A_271108 (p<0.05), and Mycoplasmoidaceae (p<0.05) were significantly increased; in the CMR group, Desulfovibrionaceae (p<0.01), Desulfovibrionales (p<0.05), Desulfovibrionia (p<0.05), Desulfoobacterota_I (p<0.01), Mailhella (p<0.01), Clostridia_258483 (p<0.05), Firmicutes_A (p<0.05), Lachnospirales (p<0.01), Lachnospirae (p<0.01), and Kineothrix (p<0.01) were significantly increased; in the CYR group, Peptostreptococcaceae_256921 (p<0.05) and Peptostreptococcales (p<0.05) were significantly increased. The above LEfSe analysis revealed significant differences in the gut microbiota among the three groups: the mycoplasma-related microbiota was significantly increased in the CCR group, the Desulfovibrio- and Clostridia-related microbiota were significantly increased in the CMR group, and the anaerobic coccus-related microbiota was significantly increased in the CYR group. These test and analysis results further confirmed the regulatory effect of Coriolus versicolor polysaccharide on the gut microbiota and its potential mechanism.

[0074] (5) As Figure 10 shown, the T-test analysis showed that: (i) From Figure 10 the phylum-level analysis in a, it can be seen that Firmicutes_A (P<0.01) and Firmicutes_B370539 (P<0.05) in the CMR group of mice were significantly higher than those in the CCR group and the CYR group. Desulfobacterota_I in the CCR group of mice was significantly lower than that in the CMR group (P<0.05) and the CYR group (P<0.05). Firmicutes_C in the CYR group of mice was significantly lower than that in the CCR group (P<0.01) and the CMR group (P<0.05). Deferribacterota in the CMR group of mice was significantly higher than that in the CYR group (P<0.05). (ii) From Figure 10Horizontal analysis at the genus level showed that Mailhella in the CCR group of mice was significantly lower than that in the CMR group (p < 0.05) and the CYR group (p < 0.05). Lawsonibacter (p < 0.05), Anaerotruncus (p < 0.05), Angelakisella (p < 0.05), Merdisoma (p < 0.05), and Allobaculum (p < 0.05) in the CMR group of mice were significantly higher than those in the CCR group and the CYR group. Mucispirillum (p < 0.05), Prevotella (p < 0.05), Tidjanibacter (p < 0.05), and Choladousia (p < 0.05) in the CMR group of mice were significantly higher than those in the CYR group. OLB9 (p < 0.05), Enterenecus (p < 0.05), Odoribacter_865974 (p < 0.05), CAG-95 (p < 0.05), Acutalibacter (p < 0.05), and Cupidesulfovibrio (p < 0.05) in the CMR group were significantly higher than those in the CCR group. Gemella (p < 0.05) and CAG-83 (p < 0.05) in the CYR group of mice were significantly higher than those in the CCR group, and Veillonella_A (p < 0.05) was significantly lower than that in the CCR group of mice. Adlercreutzia_404257 and Acetilactobacillus (p < 0.05) in the CYR group of mice were significantly higher than those in the CCR group (p < 0.05) and the CMR group (p < 0.05)

[0075] The above T-test analysis demonstrated significant differences in the intestinal flora at the phylum and genus levels among the three groups. Firmicutes A and B370539 in the CMR group were significantly increased, while Desulfobacterota I was significantly decreased; Firmicutes C in the CYR group was significantly decreased, but Anaerococcaceae and Anaerococculales were significantly increased. At the genus level, multiple genera in the CMR group were significantly increased, while certain specific genera such as Gemella and CAG-83 in the CYR group were significantly increased, indicating that the polysaccharide of Coriolus versicolor has a regulatory effect on the intestinal flora.

[0076] Example 4

[0077] This example was an experiment on the effects of the Tibetan medicine (polysaccharide powder of Coriolus versicolor) prepared in Example 1 above on the metabolites and their pathways of mice.

[0078] Thirty 4-week-old BALB / C mice with a body weight of 23.18 ± 1.26 g were selected and randomly divided into three groups of 10 mice each, namely the CCR group (blank control group), the CMR group (potassium dichromate modeling group), and the CYR group (polysaccharide of Coriolus versicolor treatment group). After 3 days of adaptive feeding, all mice started the formal experiment. The entire experimental period was 35 days. During the experiment, the mice in the CMR group and the CYR group were given intragastric administration of potassium dichromate (K2Cr2O7, dose of 10 mg / kg) every day. At the same time, the CYR group received intragastric administration of polysaccharide of Coriolus versicolor (dose of 25 mg / kg) for treatment, while the CCR group and the CMR group received intragastric administration of an equal amount of sterile water as a control. All mice were housed under a 12-hour light / dark cycle and had free access to food and water.

[0079] At the end of the experiment on the 36th day, all the mice in the above groups were euthanized by inhaling carbon dioxide. Subsequently, liver samples of each group of mice were collected for metabolomics analysis to explore the functions of metabolites and the changes in their related pathways under different treatment conditions. To further understand the relationship between the physiological indexes of mice, bacterial abundance, and metabolites, a correlation analysis was also performed by the Spearman method. The specific test and analysis results are as follows.

[0080] (1) As Figure 11 shown, (i) From the KEGG functional analysis in Figure 11 a, it can be seen that the mice in the CMR group showed inhibition of the functions of microbial membrane transduction, translation, replication and repair, and lipid metabolism, while promoting the metabolism of terpenoids and polyketides, cofactors and vitamins, and amino acids; while the mice in the CYR group treated with polysaccharide of Coriolus versicolor showed opposite metabolic changes to those in the CMR group, showing the restoration or reversal of related functions. (ii) From the MetaCyc analysis in Figure 11 b, it can be seen that the mice in the CMR group showed inhibition of the functions of the microbiota in vivo for nucleoside and nucleotide biosynthesis, fatty acid and lipid biosynthesis, while promoting the function of amino acid biosynthesis; while the microbiota function of the mice in the CYR group was restored, showing the normalization of related functions. (iii) From Figure 11The KEGG pathway abundances in c showed that: the abundances of aminoacyl-tRNA biosynthesis (p<0.05), bacterial secretion system (p<0.01), D-alanine metabolism (p<0.05), nucleotide excision repair (p<0.05), protein export (p<0.05), toluene degradation (p<0.05) and Vibrio cholerae pathogenic cycle (p<0.05) were significantly higher in the CCR group; the abundances of D-arginine and D-ornithine metabolism (p<0.01), phenylalanine metabolism (p<0.05), RNA polymerase (p<0.05), tropane, piperidine and pyridine alkaloid biosynthesis (p<0.01) and β-lactam resistance (p<0.01) were significantly higher in the CMR group of mice; while the abundance of RNA degradation was significantly higher in the CYR group (p<0.05). (ⅳ) From Figure 11The abundances of MetaCyc pathways in d showed that: in the CCR group, the abundances of COA-PWY (p<0.05), HEME-BIOSYNTHESIS-II (p<0.05), PEPTIDOGLYCANSYN-PWY (p<0.05), PHOSLIPSYN-PWY (p<0.05), POLYISOPRENSYN-PWY (p<0.01), PPGPPMET-PWY (p<0.05), PWY-5686 (p<0.05), PWY-5918 (p<0.05), PWY-6151 (p<0.05), PWY-6387 (p<0.05), PWY-6876 (p<0.01) and REDCITCYC (p<0.05) were relatively high; in the CMR group, the abundances of CENTFERM-PWY (p<0.05), COBALSYN-PWY (p<0.05), P163-PWY (p<0.05), P164-PWY (p<0.05), PWY-5005 (p<0.05), PWY-5177 (p<0.05), PWY-5304 (p<0.01), PWY-5505 (p<0.01), PWY-5509 (p<0.05), PWY-5676 (p<0.05), PWY-5677 (p<0.01), PWY-6269 (p<0.05), PWY-6478 (p<0.01), PWY-6588 (p<0.05), PWY-6590 (p<0.05), PWY-6608 (p<0.05), PWY-7013 (p<0.05), PWY-7090 (p<0.01), PWY-7198 (p<0.05), PWY-7210 (p<0.05), PWY0-1479 (p<0.05), PWY0-781 (p<0.05), PYRIDNUCSAL-PWY (p<0.01), SALVADEHYPOX-PWY (p<0.05), sulte-cys-pwy (p<0.05) were relatively high; in the CYR group, the abundances of ARGORNPROST-PWY (p<0.05), GLYCOLYSIS-E-D (p<0.05), PWY-7187 (p<0.05), PWY-7221 (p<0.05), so4sim-pwy (p<0.05) were relatively high.

[0081] Through the functional and pathway abundance analysis of KEGG and MetaCyc, it can be seen that under the induction of potassium dichromate, a series of changes occurred in the metabolic function of mice in the CMR group, including the inhibition and promotion of certain metabolic pathways. In the CYR group of mice treated with Coriolus versicolor polysaccharide, their metabolic function was restored to a certain extent, as manifested by the significant differences in the abundance of related pathways compared with the CCR group and the CMR group. This indicates that Coriolus versicolor polysaccharide may have a potential protective effect on the metabolic disorders caused by potassium dichromate by regulating the intestinal flora and its metabolites.

[0082] (2) As Figure 12 shown, a total of 3391 metabolites were identified in mice, of which 2163 were positive metabolites and 1395 were negative metabolites. By comparing the metabolite abundances of mice in the CCR group, CMR group, and CYR group, significant differences were found among the three groups. (i) From Figure 12 the number of differential metabolites in a, it can be seen that there were 35 metabolites with significant differences between the CCR group and the CMR group, 189 metabolites with significant differences between the CMR group and the CYR group, and 180 metabolites with significant differences between the CCR group and the CYR group; at the same time, the representative differential metabolites between CCR group mice and CMR group mice were: Dazepam (p < 0.001), Erionyl Yellow AR (p < 0.01), 2-Aminoisobutyric acid (p < 0.01), 2-Aminobutyric acid (p < 0.01), Dimethylglycine (p < 0.01), and 4-Aminobutyric acid (GABA) (p < 0.01). The typical differential metabolites between CMR group mice and CYR group mice were: Glucoiberin (p < 0.0001), M413T284 (p < 0.0001), Cellopentaose (p < 0.0001), Erionyl Yellow AR (p < 0.001), and 3-β-Glucosyltrisaccharide (p < 0.0001). The main differential metabolites between CCR group mice and CYR group mice were: N-Acetylgalactosamine (p < 0.0001), N-Acetylglucosamine (p < 0.0001), 5-Amino-2-(5-amino-1,3-benzothiazol-2-yl)phenol (p < 0.0001), 2-Benzylmalic acid (p < 0.0001), and N-Acetylmannosamine (p < 0.001). (ii) From Figure 12From the changes in the abundances of specific metabolites in b, it can be seen that the abundances of N-(1H-benzimidazol-2-yl)-3-(4-fluorophenyl)-1H-pyrazole-4-carboxamide (p < 0.05), cephamycin C (p < 0.01), and Erionyl Yellow AR (p < 0.01) in the mice of the CMR group were significantly increased, while the abundance of N-(4-methoxyphenyl)-2-(1-piperazinyl)acetamide decreased; in the mice of the CYR group, the abundance of N-(4-methoxyphenyl)-2-(1-piperazinyl)acetamide increased, and the abundances of N-(1H-benzimidazol-2-yl)-3-(4-fluorophenyl)-1H-pyrazole-4-carboxamide (p < 0.05), cephamycin C (p < 0.01), and Erionyl Yellow AR (p < 0.01) decreased.

[0083] From the above analysis, it can be seen that Coriolus versicolor polysaccharide has an obvious regulatory effect on potassium dichromate-induced metabolic disorders in mice. The changes in the abundances of various metabolites in the mice of the CYR group are opposite to those of the CMR group, and the abundances of some metabolites are restored to levels close to those of the CCR group. This indicates that Coriolus versicolor polysaccharide may have a potential therapeutic effect on metabolic abnormalities caused by potassium dichromate by regulating the expression of specific metabolites.

[0084] (3) As Figure 13 shown, through KEGG pathway analysis, it was found that there were significant differences in taste transduction (p < 0.05), estrogen signaling pathway (p < 0.05), gonadotropin-releasing hormone secretion (p < 0.05), and quorum sensing (p < 0.05) between the mice of the CCR group and the CMR group; there were significant differences in starch and sucrose metabolism (p < 0.01), nucleotide sugar biosynthesis (p < 0.01), amino sugar and nucleotide sugar metabolism (p < 0.01), phosphotransferase system (PTS) (p < 0.01), O-antigen nucleotide sugar biosynthesis (p < 0.01), carbohydrate digestion and absorption (p < 0.01), and glucagon signaling pathway (p < 0.05) between the mice of the CMR group and the CYR group; between the mice of the CCR group and the CYR group, there were significant differences in phosphotransferase system (PTS) (p < 0.0001), starch and sucrose metabolism (p < 0.0001), carbohydrate digestion and absorption (p < 0.001), ABC transporters (p < 0.01), galactose metabolism (p < 0.01), nucleotide sugar biosynthesis (p < 0.05), and amino sugar and nucleotide sugar metabolism (p < 0.05) pathways.

[0085] Through the above KEGG pathway analysis, it can be seen that there are significant differences in multiple metabolic pathways among the mice in the CCR group, CMR group, and CYR group; under the induction of potassium dichromate, a series of changes occurred in the metabolic function of the mice in the CMR group, while in the CYR group of mice treated with Coriolus versicolor polysaccharide, their metabolic function was restored to a certain extent; this indicates that Coriolus versicolor polysaccharide may have a potential therapeutic effect on the metabolic abnormalities caused by potassium dichromate by regulating the expression of specific metabolic pathways.

[0086] The above has introduced in detail a Tibetan medicine for relieving liver injury caused by potassium dichromate, its preparation method and application disclosed in the present invention. Specific examples are used in this article to elaborate on the principle and implementation manner of the present invention. The description of the above embodiments is only used to help understand the method and its core idea of the present invention. It should be noted that for those of ordinary skill in the art of this technology, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A method for preparing a Tibetan medicine for alleviating liver damage caused by potassium dichromate, characterized in that: The following steps are involved: (1) Grinding an appropriate amount of dried Tibetan Coriolus versicolor fruiting bodies into coarse powder, adding the Tibetan Coriolus versicolor coarse powder into a 95% ethanol solution, ultrasonically treating the solution in an ultrasonic cleaner for 30 minutes, and filtering the solution to obtain a mixture; (2) adding sterile water to the mixture obtained in step (1) for extraction, repeating this process twice, and combining the water extracts obtained from the two extractions to obtain an extract; (3) deproteinizing the extract obtained in step (2) using Sevag reagent, allowing the solution to stand for stratification after being fully mixed with the Sevag reagent, removing the protein layer, retaining the aqueous phase containing the crude polysaccharide, and obtaining a crude polysaccharide solution; (4) pouring the crude polysaccharide solution obtained in step (3) into a dialysis bag with a molecular weight cutoff value of 3500Da, placing it in a large amount of sterile water for 48 hours, and after the dialysis is completed, collecting the polysaccharide solution in the dialysis bag, freeze-drying it using a freeze dryer to obtain Tibetan Coriolus versicolor polysaccharide powder, that is, preparing a Tibetan medicine that can alleviate liver damage caused by potassium dichromate.

2. The method for preparing a Tibetan medicine for alleviating liver damage caused by potassium dichromate according to claim 1, characterized in that: In step (1), the mass volume ratio of the Tibetan Coriolus coarse powder to the 95% ethanol solution is 1g: (15-30)mL.

3. The method for preparing a Tibetan medicine for alleviating liver damage caused by potassium dichromate according to claim 1, characterized in that: In step (2), the mass volume ratio of the mixture to sterile water is 1g: (8~16)mL; the extraction temperature is 100℃ and the extraction time is 90min.

4. The method for preparing a Tibetan medicine for alleviating liver damage caused by potassium dichromate according to claim 1, characterized in that: In step (3), the volume ratio of chloroform to n-butanol in the Sevag reagent is 4:

1.

5. The method for preparing a Tibetan medicine for alleviating liver damage caused by potassium dichromate according to claim 4, characterized in that: In step (4), the freeze-drying temperature is -80°C and the freeze-drying time is 48 hours.

6. A Tibetan medicine prepared by the preparation method according to any one of claims 1 to 5 and capable of alleviating liver damage caused by potassium dichromate.

7. Use of the Tibetan medicine according to claim 6 in preparing a pharmaceutical product having the effect of alleviating liver damage.

8. The use according to claim 7, characterized in that: The pharmaceutical product for alleviating liver damage is a pharmaceutical product for alleviating or treating liver damage caused by potassium dichromate.

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