Application of plant extract in preparation of medicine for treating hepatic encephalopathy

Abstract

The invention relates to the technical field of biology, and discloses an application of a plant extract in preparation of a medicine for treating hepatic encephalopathy, the plant extract is a mixture of curcumin and epigallocatechin gallate, and the plant extract activates oxidative stress by inhibiting or preventing TXNIP from passing through a PI3K / Akt signal channel, so that the hepatic encephalopathy can be treated. Further, the induction of excessive autophagy of astrocytes is prevented, so that the occurrence or development of hepatic encephalopathy is prevented. When the plant extract is used for treating hepatic encephalopathy, the curative effect of the medicine can be enhanced by adopting the plant extract, a new treatment thought is provided, and the plant extract has important significance for improving prognosis of patients.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to use of a plant extract in preparing a medicine for treating hepatic encephalopathy. Background Art

[0002] Hepatic encephalopathy (HE), also known as hepatic coma, is a common and complex neuropsychiatric syndrome complicated by end-stage liver disease and the most common cause of death in advanced liver cirrhosis. HE is caused by brain dysfunction caused by acute or chronic liver failure or portal vein shunt, and manifests as neurological or psychological abnormalities, elevated brain ammonia levels, impaired consciousness, abnormal behavior, and even coma. Excessive ammonia and inflammation synergistic effects lead to astrocyte swelling and brain edema as its common characteristics.

[0003] Studies have shown that PI3K / Akt is involved in the fatty acid metabolism pathway of glioblastoma cells. In the absence of cholesterol and fatty acids, PI3K / Akt can activate SREBP-1 / Fas signaling elements, upregulate low-density lipoprotein receptors, and enhance fatty acid synthesis. SLC25A32 promotes the malignant progression of glioblastoma by activating the PI3K-AKT signaling pathway to enhance tumor cell migration and invasion. NQO1 drives the invasiveness of glioblastoma cells by inducing epithelial-mesenchymal transition through the PI3K / Akt / mTOR pathway. With the continuous deepening of research on the PI3K / Akt signaling pathway, it has been found that it is not only involved in the occurrence and development of a variety of diseases, but there are few reports on TXNIP activating oxidative stress through the PI3K / Akt signaling pathway, inducing astrocyte autophagy, and affecting the occurrence and development of hepatic encephalopathy.

[0004] Curcumin is a polyphenol compound extracted from the rhizome of turmeric (Curcuma longa) with multiple pharmacological effects, including antioxidant, anti-inflammatory, anti-cancer, neuroprotective, cardiovascular protection, etc. Curcumin has been proven to have antioxidant and anti-inflammatory effects and can inhibit the expression of multiple cytokines and enzymes, but there are currently no reports on the use of curcumin in the treatment of hepatic encephalopathy. Summary of the invention

[0005] The purpose of the present invention is to provide a use of a plant extract in preparing a medicine for treating hepatic encephalopathy.

[0006] In view of this, the technical solution of the present invention is as follows:

[0007] A use of a plant extract in preparing a medicine for treating hepatic encephalopathy, wherein the plant extract is a mixture of curcumin and epigallocatechin gallate.

[0008] Furthermore, the plant extract inhibits or prevents TXNIP from activating oxidative stress through the PI 3K / Akt signaling pathway, thereby preventing the induction of excessive autophagy in astrocytes, thereby preventing the occurrence or development of hepatic encephalopathy.

[0009] Furthermore, the plant extract is prepared by the following method:

[0010] Wash and slice the turmeric rhizome, dry it naturally or use low-temperature drying at 40-60°C until the moisture content drops below 10%, and then grind it into fine powder with a particle size of 100-200 mesh to obtain turmeric powder;

[0011] Using 70% ethanol as a solvent, mixing turmeric powder and the solvent in a ratio of 1:10, putting the mixture into an extraction tank, performing ultrasonic extraction or reflux extraction, filtering and collecting the filtrate, and putting the filtrate into a rotary evaporator to evaporate and remove the solvent to obtain curcumin;

[0012] The curcumin and epigallocatechin gallate are mixed in a ratio of 10:3 to obtain the plant extract.

[0013] Further, the ultrasonic extraction is ultrasonic treatment at 40-50°C and 100-150W for 30-60min;

[0014] The reflux extraction is to heat the mixture of the turmeric powder and the solvent in the extraction tank to boiling and perform constant temperature reflux extraction for 1-2 hours.

[0015] The present invention also provides a plant extract prepared according to the above scheme.

[0016] The present invention also provides a medicine for treating hepatic encephalopathy, wherein the medicine is composed of curcumin and epigallocatechin gallate or contains curcumin and epigallocatechin gallate.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The mixture of curcumin and epigallocatechin gallate can inhibit the activation of PI 3K / Akt signaling pathway by regulating the expression of TXNIP. TXNIP is an important factor regulating oxidative stress. It activates the PI3K / Akt signaling pathway, leading to increased oxidative stress and promoting cell damage. Inhibiting oxidative stress can reduce the production of oxygen free radicals, reduce the risk of intracellular redox imbalance, and alleviate neurological damage in patients with hepatic encephalopathy.

[0019] Astrocytes play an important role in hepatic encephalopathy, and their dysfunction is closely related to pathological progression. The combination of curcumin and epigallocatechin gallate can reduce the level of autophagy in astrocytes by inhibiting the activation of TXNIP. Preventing excessive autophagy in astrocytes can maintain the normal metabolism and function of cells, reduce the risk of apoptosis and cerebral edema, and help protect brain tissue.

[0020] Curcumin also has antioxidant, anti-inflammatory, anti-tumor and neuroprotective effects, and can inhibit the production of inflammatory factors and reduce inflammatory responses. As a strong antioxidant, epigallocatechin gallate can effectively scavenge free radicals and enhance the antioxidant capacity of cells.

[0021] The present invention emphasizes the role of TXNIP in the occurrence and development of hepatic encephalopathy, provides a new target, and expands the treatment strategy for hepatic encephalopathy. By directly regulating TXNIP and PI 3K / Akt pathways, it can provide a new direction for drug development. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Various other advantages and benefits will become apparent to those of ordinary skill in the art by reading the detailed description of the preferred embodiments below. The accompanying drawings are only for the purpose of illustrating the preferred embodiments and are not to be considered as limiting the present invention. Moreover, the same reference symbols are used throughout the accompanying drawings to represent the same components. In the accompanying drawings:

[0023] Figure 1 This is a diagram of HE related data analysis results in the GEO database of an embodiment of the present invention;

[0024] Figure 2 This is a volcano plot of differentially expressed genes in an embodiment of the present invention;

[0025] Figure 3 GO biological process enrichment map of differentially expressed genes in the embodiments of the present invention;

[0026] Figure 4 This is a KEGG pathway enrichment map of the differentially expressed genes in the examples of the present invention. DETAILED DESCRIPTION

[0027] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0028] Thioredoxin interacting protein (TXNIP), also known as TBP2, is an important regulator of oxidative stress in the body. Its cysteine ​​residue at position 247 can form a stable disulfide bond with the cysteine ​​residue at position 32 of thioredoxin, thereby negatively regulating the expression of the antioxidant factor thioredoxin and affecting the body's oxidation-antioxidation capacity. Studies have found that TXNIP mainly participates in various cellular processes, including regulating oxidative stress, stimulating angiogenesis, and promoting inflammation, by binding to the active cysteine ​​residues of reduced TRX. It can also participate in regulating signaling pathways in different cells independently of the TRX system through non-redox-dependent effects, including cell apoptosis, stimulating insulin secretion, and regulating cell growth. As a multifunctional protein, TXNIP plays an important regulatory role in many diseases such as diabetes, cardiovascular disease, breast cancer, Alzheimer's disease, and Parkinson's disease. For example, metformin reduces oxidative stress and inflammatory response through the TXNIP-NLRP3-GSDMD pathway, preventing intestinal ischemia-reperfusion injury and cell pyroptosis. Silencing TXNIP regulates the progression of ischemic stroke by alleviating oxidative stress damage through the MAPK-NRF2 axis. Animal experiments have confirmed that cognitive decline, oxidative damage in the brain and spleen, and the production of inflammatory cytokines in Alzheimer's disease mouse models are all related to the expression and regulation of TXNIP. High expression of TXNIP can induce the proinflammatory phenotype and caspase-3-dependent apoptosis of astrocytes. The above studies suggest that TXNIP may cause brain dysfunction by activating oxidative stress, thereby participating in the progression of HE, but its specific mechanism of action is still unclear. The applicant has been committed to the study of HE for many years. In the early stage, bioinformatics analysis was performed based on HE-related data in the GEO database, and it was found that the expression level of TXNIP was significantly upregulated in HE samples. Further pathway enrichment analysis of differentially expressed genes was performed, and the results showed that the differentially expressed genes were significantly enriched in the PI 3K / Akt signaling pathway. The results of in vivo animal experiments also showed that TXNIP and PI 3K / Akt signaling pathway-related protein PI 3K were significantly overexpressed in HE mouse models. The results of GEO data set analysis found that TXNIP was significantly overexpressed in HE and may be involved in HE progression through the PI 3K-Akt signaling pathway. Based on the transcriptome sequencing data of GSE230287 in the GEO database, the differentially expressed genes in the sham operation group and the HE model group were analyzed. The results showed that there were 398 differentially expressed genes between the two groups, among which TXNIP was significantly overexpressed in the HE group, such as Figure 1 , Figure 2 Further GO biological process and KEGG pathway enrichment analysis of the differentially expressed genes showed that the differentially expressed genes were highly enriched in the PI 3K-Akt signaling pathway, such as Figure 3 , Figure 4As shown. This suggests that TXNIP may be involved in the progression of HE through the PI 3K-Akt signaling pathway. It is proposed that TXNIP activates oxidative stress through the PI 3K / Akt signaling pathway, thereby inducing excessive autophagy in astrocytes and promoting the occurrence and development of hepatic encephalopathy. In other words, inhibiting TXNIP from activating oxidative stress through the PI 3K / Akt signaling pathway can inhibit or prevent the occurrence and development of hepatic encephalopathy.

[0029] The following will be explained by specific examples. The epigallocatechin gallate in the following examples is purchased from Sichuan Jingcuibao Biotechnology Co., Ltd.

[0030] Example 1

[0031] Preparation of plant extracts and medicines for treating hepatic encephalopathy

[0032] Step 1: Raw material preparation

[0033] Turmeric rhizome processing: Select 500 grams of fresh turmeric rhizome and wash it thoroughly with clean water. Cut the washed turmeric rhizome into thin slices with a thickness of about 2-3 mm to facilitate subsequent drying and extraction.

[0034] Step 2: Drying

[0035] Drying: Low-temperature drying is performed in an oven at 60°C until the moisture content drops below 10%, which takes about 24-36 hours.

[0036] Step 3: Crushing

[0037] Grinding turmeric: Grind the dried turmeric flakes into fine powder, the particle size of the powder is controlled at 100-200 mesh, and about 200 grams of turmeric powder is obtained.

[0038] Step 4: Extract Curcumin

[0039] Extraction process:

[0040] Solvent preparation: Use 2000 ml of 70% ethanol as the solvent.

[0041] Mixing ratio: Mix 200 g of turmeric powder with 2000 ml of 70% ethanol in a ratio of 1:10.

[0042] Ultrasonic extraction: Ultrasonic extraction was performed at 120 W at 45 °C for 45 min to ensure uniform extraction.

[0043] Filtration: After the extraction is completed, filter the extract and collect the filtrate.

[0044] Step 5: Evaporation and concentration

[0045] Removing the solvent: The collected filtrate was placed in a rotary evaporator and evaporated to remove the solvent at 50°C until about 100 g of concentrated curcumin was obtained.

[0046] Step 6: Mix the plant extracts

[0047] Preparation of plant extract: 100 g of the obtained curcumin and 30 g of epigallocatechin gallate (EGCG) were mixed in a ratio of 10:3, and stirred thoroughly to obtain the final plant extract.

[0048] Step 7: Drug preparation

[0049] Drug formula: The plant extract is used to prepare a drug for treating hepatic encephalopathy. It can be mixed with 50 grams of starch, 10 grams of microcrystalline cellulose and 5 grams of talc to form tablets, which are compressed by a tablet press to obtain about 200 tablets of the drug, each containing the active ingredients of curcumin and epigallocatechin gallate.

[0050] Example 2

[0051] Preparation of plant extracts and medicines for treating hepatic encephalopathy

[0052] Step 1: Raw material preparation

[0053] Turmeric rhizome processing: Select 500 grams of fresh turmeric rhizome and wash it thoroughly with clean water. Cut the washed turmeric rhizome into thin slices with a thickness of about 2-3 mm to facilitate subsequent drying and extraction.

[0054] Step 2: Drying

[0055] Drying: Low-temperature drying is performed in an oven at 60°C until the moisture content drops below 10%, which takes about 24-36 hours.

[0056] Step 3: Crushing

[0057] Grinding turmeric: Grind the dried turmeric flakes into fine powder, the particle size of the powder is controlled at 100-200 mesh, and about 200 grams of turmeric powder is obtained.

[0058] Step 4: Extract Curcumin

[0059] Extraction process:

[0060] Solvent preparation: Use 2000 ml of 70% ethanol as the solvent.

[0061] Mixing ratio: Mix 200 g of turmeric powder with 2000 ml of 70% ethanol in a ratio of 1:10.

[0062] Reflux extraction:

[0063] The mixture in the extraction tank was heated to boiling and kept under constant temperature reflux for 1.5 hours.

[0064] Filtration: After the extraction is completed, filter the extract and collect the filtrate.

[0065] Step 5: Evaporation and concentration

[0066] Removing the solvent: The collected filtrate was placed in a rotary evaporator and evaporated to remove the solvent at 50°C until about 100 g of concentrated curcumin was obtained.

[0067] Step 6: Mix the plant extracts

[0068] Preparation of plant extract: 100 g of the obtained curcumin and 30 g of epigallocatechin gallate (EGCG) were mixed in a ratio of 10:3, and stirred thoroughly to obtain the final plant extract.

[0069] Step 7: Drug preparation

[0070] Drug formula: The plant extract is used to prepare a drug for treating hepatic encephalopathy. It can be mixed with 50 grams of starch, 10 grams of microcrystalline cellulose and 5 grams of talc to form tablets, which are compressed by a tablet press to obtain about 200 tablets of the drug, each containing the active ingredients of curcumin and epigallocatechin gallate.

[0071] Experimental design

[0072] Animal model: 30 adult male Wistar rats weighing approximately 200-250 g were randomly divided into three groups (10 rats in each group):

[0073] Control group: normal diet, no treatment.

[0074] Model group: A hepatic encephalopathy model was established by inducing hepatic encephalopathy using amino acid ammonia water (100 mg / kg).

[0075] Experimental group: divided into two groups (experimental group 1 and experimental group 2). On the basis of the model group, each group was given the plant extract obtained in Examples 1 and 2 (100 mg / kg feed per day) for treatment.

[0076] Experimental procedures

[0077] Hepatic encephalopathy model establishment:

[0078] Hepatic encephalopathy was induced by injection of amino acid ammonia (100 mg / kg). The behavioral changes and clinical symptoms of mice were observed.

[0079] Treatment process:

[0080] The control group was fed normally, and the model group and the experimental group were treated on the first day after induction for 14 days. The experimental group was given plant extracts daily at a dose of 100 mg / kg.

[0081] Behavioral testing:

[0082] On the 7th and 14th days of treatment, the cognitive and motor abilities of the rats were evaluated using a maze test, which was repeated three times. The results are shown in Table 1.

[0083] Table 1

[0084]

[0085] As can be seen from the above table: the completion time of the rats in the model group increased significantly, especially in the test on the 14th day, the completion time was significantly higher than that on the 7th day, indicating that hepatic encephalopathy had a serious impact on their cognitive and motor abilities. This phenomenon is related to ammonia-induced hepatic encephalopathy, which leads to impaired neurological function and slow response, further affecting the learning ability and memory of rats. This result supports the effectiveness of the hepatic encephalopathy model and shows its negative impact on animal behavior. The performance of the experimental group was improved relative to the model group. Experimental Group 1 and Experimental Group 2 showed significant improvement compared with the 7th day in the test on the 14th day, especially Experimental Group 2, whose completion time was significantly reduced on the 14th day, showing a performance close to that of the control group. This shows that the plant extract reversed the ammonia-induced cognitive deficits to a certain extent, possibly by improving neurological function or promoting neuroprotective mechanisms.

[0086] Overall, the data support the potential role of plant extracts in improving cognitive abilities in rats with hepatic encephalopathy. The significant cognitive decline in the model group contrasts sharply with the improvement in the experimental group, further verifying the mechanism of hepatic encephalopathy's effects on behavior. At the same time, this result provides a basis for further research on plant extracts. In the future, we can explore their specific mechanisms of action and active ingredients, and provide new ideas and directions for the treatment of hepatic encephalopathy.

[0087] Biochemical testing:

[0088] After the treatment, blood samples were taken from rats in each group to detect the following indicators in serum:

[0089] Liver function: Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels.

[0090] Oxidative stress indicators: malondialdehyde (MDA), superoxide dismutase (SOD) activity, the results are shown in Table 2.

[0091] Table 2

[0092] Control group Model Group Experimental Group 1 Experimental Group 2 ALT, U / L 29 88 68 49 AST, U / L 36 110 81 63 MDA, μmol / L 2.4 6.6 4.4 3.2 SOD, U / mL 189 79 121 149

[0093] As shown in Table 2: Analysis of liver function indicators: ALT and AST levels in the control group were low, indicating normal liver function. ALT and AST levels in the model group increased significantly, indicating liver cell damage. ALT and AST levels in experimental groups 1 and 2 were lower than those in the model group, especially in experimental group 2, which was close to the control group level. This shows that plant extracts can effectively improve liver cell damage, suggesting that they may have a liver protective effect.

[0094] The MDA level in the control group was low, indicating that the oxidative stress level was normal. The MDA level in the model group was significantly increased, indicating that hepatic encephalopathy led to increased oxidative stress. The MDA levels in experimental groups 1 and 2 were lower than those in the model group, especially in experimental group 2, indicating the potential role of plant extracts in reducing oxidative stress.

[0095] The SOD activity of the control group was higher, indicating that it had strong antioxidant capacity. The SOD activity of the model group was significantly reduced, indicating that the antioxidant capacity decreased and oxidative stress increased. The SOD activity of experimental groups 1 and 2 was restored, especially in experimental group 2, whose activity was close to that of the control group. This suggests that plant extracts may improve liver function and antioxidant capacity by increasing SOD activity.

[0096] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. Use of a plant extract in the preparation of a medicament for treating hepatic encephalopathy, characterized in that: The plant extract is a mixture of curcumin and epigallocatechin gallate.

2. Use of a plant extract according to claim 1 in preparing a drug for treating hepatic encephalopathy, characterized in that: The plant extract inhibits or blocks TXNIP from activating oxidative stress through the PI 3K / Akt signaling pathway, thereby preventing the induction of excessive autophagy of astrocytes, thereby preventing the occurrence or development of hepatic encephalopathy.

3. Use of a plant extract according to claim 1 in preparing a drug for treating hepatic encephalopathy, characterized in that: The plant extract is prepared by the following method: Wash and slice the turmeric rhizome, dry it naturally or use low-temperature drying at 40-60°C until the moisture content drops below 10%, and then grind it into fine powder with a particle size of 100-200 mesh to obtain turmeric powder; Using 70% ethanol as a solvent, mixing turmeric powder and the solvent in a ratio of 1:10, putting the mixture into an extraction tank, performing ultrasonic extraction or reflux extraction, filtering and collecting the filtrate, and putting the filtrate into a rotary evaporator to evaporate and remove the solvent to obtain curcumin; The curcumin and epigallocatechin gallate are mixed in a ratio of 10:3 to obtain the plant extract.

4. Use of a plant extract according to claim 3 in preparing a medicament for treating hepatic encephalopathy, characterized in that: The ultrasonic extraction is performed at 40-50°C and 100-150W for 30-60min; The reflux extraction is to heat the mixture of the turmeric powder and the solvent in the extraction tank to boiling and perform constant temperature reflux extraction for 1-2 hours.

5. A plant extract, characterized in that The method is prepared according to the use of a plant extract according to any one of claims 1 to 4 in preparing a medicament for treating hepatic encephalopathy.

6. A drug for treating hepatic encephalopathy, characterized in that: It consists of or contains curcumin and epigallocatechin gallate.

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