Application of astragalus extract

Abstract

The invention discloses an application of an astragalus extract, and particularly relates to an application of the astragalus extract in preparation of a medicine for treating heatstroke and a medicine for treating heatstroke combined with liver injury. The invention provides a new application of the astragalus extract in the field of treating severe heatstroke / heatstroke combined liver injury, and it is clear that the astragalus extract preparation has a good effect of improving severe heatstroke, can significantly improve the in-vivo environmental homeostasis of an organism, alleviates tissue injury caused by heat exposure, and has a good application prospect in the field of treating severe heatstroke / heatstroke combined liver injury. Meanwhile, the astragalus extract preparation can also effectively relieve the acute liver injury complicated by the heatstroke and can be used as a potential medicine for treating the liver injury complicated by the heatstroke. The radix astragali extract has dual protection effects in treatment of severe heatstroke and related liver injury, and provides a new intervention strategy for treatment of heat stress related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of an astragalus extract in preparing a drug for treating severe heatstroke or heatstroke combined with liver damage and enhancing the intestinal mucosal barrier function. Background Art

[0002] The global climate system is undergoing unprecedented and dramatic changes. Observational data indicate that the trend of global warming continues to intensify, directly leading to the significant increase in the frequency, intensity, and duration of extreme weather and climate events. These extreme events are often characterized by large-scale and persistent occurrences, posing multi-dimensional and complex threats to natural ecosystems, socioeconomic operations, and human health. The latest monitoring report from the World Meteorological Organization (WMO) reveals that in 2024, multiple indicators of the global climate system exceeded historical extremes: the annual mean near-surface temperature rose by 1.55±0.13°C compared to the pre-industrial baseline (1850-1900), marking the first year in which temperatures consistently exceeded the 1.5°C threshold, setting the record for the warmest year since systematic observations began in 1849. This warming trend is reshaping the characteristic patterns of extreme heat waves worldwide, leading to an increasingly severe public health crisis and an exponential increase in heat-related mortality. Heat-related health risks have become one of the most significant global health challenges of this century.

[0003] Heatstroke is a critical syndrome induced by extreme high temperature environment. Its pathological characteristics are mainly manifested as: a sharp increase in core temperature (>40℃), abnormalities in the central nervous system (manifested as impaired consciousness, convulsions or coma), and damage to multiple organ functions (typically the liver and intestines). Its pathogenesis involves three pathophysiological processes: direct cell damage caused by heat stress, systemic inflammatory cascade and coagulation dysfunction. There are two clinical types: prodromal and mild cases (good prognosis after leaving the heat source) and severe cases (often accompanied by multiple organ dysfunction syndrome, with significantly increased mortality). The clinical treatment of heatstroke focuses on rapid cooling. There is currently no specific therapeutic drug, so it is of great significance to explore the therapeutic drugs for heatstroke.

[0004] As the core organ for metabolic regulation in the body, the liver plays a key role in lipid metabolism, glucose metabolism balance, and energy homeostasis. High temperature stress can induce significant liver pathological changes, the characteristic manifestations of which include: hepatocyte swelling, vacuolation, fat deposition, and extensive necrosis in the central area of the lobule. The progression of acute liver injury to liver failure is the main pathological basis for death from heatstroke. At the same time, high temperature exposure can lead to damage to the intestinal barrier system, and its mechanisms involve: destruction of intestinal epithelial tight junctions, abnormally increased mucosal permeability, and translocation of bacteria and endotoxins into the blood. There is a bidirectional regulation between liver disease and intestinal flora: the liver regulates the flora ecology through bile acid secretion, IgA release, and other pathways, and the metabolites of the flora (such as short-chain fatty acids) affect liver function. However, there is a lack of specific treatments for heatstroke-related liver injury. The development of therapeutic intervention strategies has significant public health value, and targeting the regulation of the gut-liver axis has become a potential intervention approach.

[0005] Astragalus is one of the most widely used basic herbs in traditional Chinese medicine. It has the effects of replenishing qi and strengthening the exterior, anti-oxidation, anti-inflammatory, regulating immunity, improving microcirculation, nourishing, protecting the liver, and diuresis. Astragalus is warm in nature, sweet in taste, and enters the lung and spleen meridians. It has the effects of replenishing qi and strengthening the exterior, and can enhance the body's immunity and adaptability. The main active ingredients in Astragalus, such as polysaccharides, saponins and flavonoids, can significantly improve the body's immune function. Astragalus can improve the damaged intestinal mucosal barrier caused by chemotherapy. However, there are no related technical inspirations or literature reports in the prior art on the application of Astragalus extracts in the treatment of severe heatstroke or acute liver damage caused by heatstroke complications and the improvement of damaged intestinal mucosal barrier caused by heatstroke. Summary of the Invention

[0006] In view of the problems existing in the prior art, the present invention aims to design and provide a technical solution for the application of Astragalus extract. The present invention is specifically implemented by the following technical solutions:

[0007] A first aspect of the present invention provides the use of an astragalus extract in the preparation of a medicine for treating heatstroke, a medicine for heatstroke combined with liver damage, and / or a medicine for enhancing the intestinal mucosal barrier function.

[0008] Furthermore, the astragalus extract in this application is used as one of the main active ingredients or the only active ingredient of the medicine.

[0009] Furthermore, the drug in this application also contains a pharmaceutically acceptable carrier.

[0010] Furthermore, the heatstroke described in this application is severe heatstroke.

[0011] Furthermore, the heatstroke combined with liver damage in this application is heatstroke combined with acute liver damage.

[0012] Furthermore, the astragalus extract in this application is prepared by the following method:

[0013] 1) Select raw Astragalus root with normal color and no moldy odor, wash it, dry it and set aside;

[0014] 2) placing the raw Astragalus root material from step 1) in an extraction tank, adding water at a mass ratio of 1:6, opening the extraction tank for heating and boiling at a temperature of 120-130° C., heating and boiling for 60 minutes, and introducing the boiled raw material juice into a double-effect concentrator for concentration and sterilization;

[0015] 3) homogenizing the concentrated and sterilized liquid obtained in step 2) in a homogenizer at a pressure of 20-25 MPa;

[0016] 4) placing the extract obtained after homogenization in step 3) in a spray drying tower for spray drying, controlling the air inlet temperature between 150-190° C. and the air outlet temperature between 80-90° C., and adjusting the feed rate according to the outlet temperature to obtain the astragalus extract.

[0017] A second aspect of the present invention provides a drug for treating heatstroke, heatstroke combined with liver damage and / or enhancing the intestinal mucosal barrier function, wherein one of the main active ingredients or the only active ingredient of the drug is an astragalus extract.

[0018] Furthermore, the drug further comprises a pharmaceutically acceptable carrier.

[0019] This invention provides a novel use of Astragalus extract in the treatment of severe heatstroke / heatstroke-associated liver damage. It also demonstrates that Astragalus extract preparations are effective in improving severe heatstroke, significantly improving the body's internal homeostasis and mitigating tissue damage caused by heat exposure. Furthermore, Astragalus extract preparations can effectively mitigate acute liver damage associated with heatstroke, making them a potential treatment for heatstroke-associated liver damage. The dual protective effects of Astragalus extract in treating severe heatstroke and its associated liver damage provide a new intervention strategy for the treatment of heat stress-related diseases.

[0020] Astragalus extract preparations enhance the intestinal mucosal barrier function, effectively preventing harmful substances such as endotoxins from entering the bloodstream through the mucosal barrier. The technology provided by this invention has significant application value in the development of drugs for severe heatstroke and its complications. This breakthrough opens up new research directions for the clinical treatment and drug development of heat-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This figure shows the effect of a single treatment with different doses of the Astragalus extract of the present invention on the core body temperature of mice;

[0022] Figure 2 Graph showing the effects of the Astragalus extract treatment on mouse liver morphology, serum alanine aminotransferase (ALT) and serum aspartate aminotransferase (AST);

[0023] Figure 3 This figure shows the effect of the Astragalus extract treatment on phosphatidic acid phosphohydrolase (Lipin 1) in mouse liver;

[0024] Figure 4 The effect of the Astragalus extract of the present invention on the intestinal mucosal barrier of mice (Note: Compared with the NC group, * P<0.05, ** P<0.01, *** P<0.001; compared with the HC group, # P<0.05,

[0025] ## P<0.01, ### P < 0.001. The t test was used to evaluate the significant differences, and the results are expressed as mean ± SEM. DETAILED DESCRIPTION

[0026] The present invention is further described below in conjunction with specific embodiments to facilitate a better understanding of the present technical solution.

[0027] Example 1: Preparation of Astragalus Extract Solution and Model Intervention Plan

[0028] Preparation of Astragalus extract: 1) selecting Astragalus raw medicinal materials with normal color and no moldy odor, washing them, drying them and setting them aside; 2) placing the Astragalus raw medicinal materials in step 1) in an extraction tank, adding water in a mass ratio of 1:6, opening the extraction tank for heating and boiling, controlling the temperature at 120-130° C., heating and boiling for 60 minutes, and introducing the boiled raw material juice into a double-effect concentration tank for concentration and sterilization; 3) homogenizing the concentrated liquid obtained after concentration and sterilization in step 2) in a homogenizer at a pressure of 20-25 MPa; 4) placing the extract obtained after homogenization in step 3) in a spray drying tower for spray drying, controlling the air inlet temperature between 150-190° C., and the air outlet temperature between 80-90° C., and adjusting the feed rate according to the outlet temperature to obtain the Astragalus extract.

[0029] The preparation of the astragalus extract solution of the present invention is based on a human oral therapeutic dose of 50g, and a single dose of 50g (50g / 70kg) is obtained for a standard body weight of 70kg. If calculated by the dosage per unit body weight, the equivalent dose of mice (standard body weight 20g) is equivalent to 9.1 times that of humans. Based on this conversion factor, the dosage of mice is 6.5g / kg (i.e., 6.5mg / g). According to the gavage volume of mice of 8μL / g, the dosage of astragalus extract in the oral dosage form is 0.8125mg / μL (0.8125g / mL) based on the final concentration in mice, i.e., 0.8125g of astragalus extract powder is required for 1mL of aqueous solution. The above is a high-dose astragalus extract preparation. The low-dose Astragalus extract solution is prepared according to the oral therapeutic dose of 25g for humans, and the single dose for a standard body weight of 70kg is 25g (25g / 70kg). The dose for mice obtained based on the above conversion coefficient is 3.25g / kg (i.e., 3.25mg / g), and the final concentration of mice is 0.40625mg / μL (0.40625g / mL), that is, 1mL of aqueous solution requires 0.40625g of Astragalus extract powder. The gavage volume for each mouse is obtained by multiplying the actual weight of the mouse by the gavage volume, and the maximum gavage volume does not exceed 200μL. In the process of optimizing the dosage of Astragalus extract, all that can achieve the technical effects described in the present invention belong to the technical solution described in the present invention and fall within the scope of protection of the present invention.

[0030] Preparation method of Astragalus extract: dissolve the Astragalus extract in ddH2O, the low dose is 3.25g / kg, the high dose is 6.5g / kg, and it is prepared and used immediately.

[0031] Model intervention plan:

[0032] Healthy male C57BL / 6N mice, 8 weeks old, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. (license number: SCXK(Zhe)2019-0001). They were housed at the Experimental Animal Center of Zhejiang Chinese Medical University at 22±1°C on a 12 / 12 hour day / night cycle and were given free access to food and water. After one week of acclimatization in an SPF animal facility, the C57BL / 6 mice were randomly divided into six groups of six mice each. The groupings were as follows:

[0033] 1) Normal control (NC)

[0034] 2) Control group (Control + Astragalus extract-low dose, CAL)

[0035] 3) Control group treated with a single high-dose Astragalus extract (CAH)

[0036] 4) Extreme high temperature exposure group (Heat control, HC)

[0037] 5) Extreme heat exposure + Astragalus extract-lowdose single treatment group (Heat + Astragalus extract-lowdose, HAL)

[0038] 6) Extreme heat exposure + Astragalus extract high dose single treatment group (Heat + Astragalus extract-high dose, HAH)

[0039] To further investigate the therapeutic effects of the single high-dose group, the CA group was designated as the control group for a single high-dose Astragalus extract treatment, and the HA group was designated as the extreme heat exposure plus a single high-dose Astragalus extract treatment. The NC, CA, HC, and HA groups served as observation groups for the therapeutic effects of the single high-dose Astragalus extract group.

[0040] To further investigate the therapeutic effects of double high-dose 6.5 g / kg Astragalus extract, 9-week-old C57BL / 6N male mice were randomly divided into three groups, with 6 mice in each group. The double treatment groups were as follows:

[0041] 1) Normal control group (Control)

[0042] 2) Extreme high temperature exposure group (Heat)

[0043] 3) Extreme heat exposure + high-dose Astragalus extract double treatment group

[0044] Single-dose Astragalus extract treatment and modeling protocol (9-hour recovery): Mice in the HC, HAL, and HAH (HA) groups were exposed to a high temperature of 39.5±0.5°C. During the exposure period, mice were deprived of food and water, and their core body temperatures were monitored using a rectal thermometer. When core body temperatures reached their maximum or when heat shock occurred, the mice in each group were released from the heat and placed in a 22±1°C environment for recovery. Heat-shocked mice in the HC group were given ddH2O, while those in the HAL group were immediately given a low-dose Astragalus extract by gavage. Heat-shocked mice in the HAH (HA) group were immediately given a high-dose Astragalus extract by gavage. All groups received food. Concurrently, mice in the CAL and CAH (CA) groups were given a single low- and high-dose Astragalus extract by gavage, respectively. Core body temperatures were continuously monitored, and mice were sacrificed after nine hours of recovery. Mice in the normal and treatment control groups were also sacrificed.

[0045] Double Astragalus extract treatment and modeling plan (Recover 24h): Mice in the Heat group and Astragalus extract group were exposed to a high temperature environment of 39.5±0.5℃. During the high temperature exposure period, the mice were deprived of water and food, and the core body temperature of the mice was monitored with a rectal thermometer. When the core body temperature of the two groups of mice reached the maximum or heat shock state, the exposure was stopped and the mice were placed in an environment of 22±1℃ for recovery. The heat-shocked mice in the Heat group were immediately given ddH2O, and the heat-shocked mice in the Astragalus extract group were immediately given high-dose Astragalus extract by gavage. After recovering for 15 hours after heat shock, the mice in the Astragalus extract group were again given high-dose Astragalus extract by gavage, and the mice in the Heat group were given ddH2O at the same time. The core body temperature of the mice continued to be monitored, and they were immediately killed after 24 hours of recovery. The normal control group was also killed at the same time.

[0046] Research plan: ① Based on the construction of the animal model in Example 1, after 9 hours and 24 hours of recovery from high temperature exposure, the mice were anesthetized with intraperitoneal injection of sodium pentobarbital, and blood was collected from the hypothalamic vena cava to measure ALT and AST. The mice in the control group were given the same treatment except for high temperature exposure. ② Detection of gene expression levels in liver tissue: Using the above-mentioned animal model, total RNA from liver samples was extracted using RNAisoPlus reagent, and RNA was reverse transcribed into cDNA using a cDNA reverse transcription kit. The mRNA level of Lipin 1 was detected using SYBR Green. ③ Detection of protein expression levels in liver tissue: Using the above-mentioned animal model, total protein from liver samples was extracted, and after measuring the concentration, the protein level of Lipin 1 was detected by Western Blot. ④ The levels of serum LPS, DAO, and D-Lactate were detected by ELISA.

[0047] Example 2: Effects of single treatment with different doses of Astragalus extract on core body temperature changes in mice;

[0048] Before exposure to extreme high temperatures, there was no difference in the core body temperature of mice in the six groups of NC, CAL, CAH, HC, HAL, and HAH. When the mice in the HC, HAL, and HAH groups reached a state of heat shock (the highest core body temperature was 41.5°C), the mice in the HC group were given pure water, the mice in the HAL group were given a low dose of Astragalus extract, and the mice in the HAH group were given a high dose of Astragalus extract. During the recovery period, when the core body temperature of the mice in the HC group reached the lowest, the lowest core body temperature of the mice in the HAH and HAL groups (31.0°C vs 29.4°C) was higher than the lowest core body temperature of the HC group (28.4°C). After nine hours of recovery (Recover 9h), the core body temperature of the mice in the HAH group had returned to near normal levels (34.6°C), while the core body temperature of the mice in the HAL and HC groups (31.8°C vs 28.3°C) had not returned to normal levels ( Figure 1 A). Further analysis of the efficacy of high-dose Astragalus extract revealed that high-dose Astragalus extract could significantly restore the core body temperature of mice for nine hours ( Figure 1 B). A single treatment with high-dose Astragalus extract significantly improved body temperature imbalance after heat shock.

[0049] Example 3: Effects of Astragalus Extract Treatment on Mouse Liver Morphology, Serum Alanine Aminotransferase (ALT) and Serum Aspartate Aminotransferase (AST);

[0050] First, we analyzed the effects of a single high-dose Astragalus extract treatment. H&E staining of the livers of mice in the normal group, the high-dose Astragalus extract control group, the heat-exposed group, and the high-dose Astragalus extract treatment group revealed abnormal liver tissue morphology, with more congestion and fat vacuoles, while a single high-dose Astragalus extract treatment significantly improved these findings ( Figure 2 A).

[0051] Subsequently, the improvement effect of single treatment with different doses of Astragalus extract on liver function was analyzed. ALT is mainly distributed in the cytoplasm of liver cells, and its serum content is extremely low under normal circumstances. When liver cells are damaged and the cell membrane permeability increases, ALT in the cytoplasm will be released into the blood in large quantities, so ALT is the most sensitive indicator of liver cell damage. AST is mainly located in the mitochondria of liver cells, and the serum basal level is also low. When severe liver damage occurs, AST will enter the blood circulation, and its degree of increase is often related to the severity of liver cell damage. After male mice recovered from extreme high temperature exposure to heat shock for nine hours, the levels of ALT and AST in the mouse serum increased significantly, indicating liver damage. Compared with the mice in the heat exposure group, single low and high doses of Astragalus extract treatment can significantly improve ALT levels, and only a single high dose of Astragalus extract treatment can significantly improve AST levels ( Figure 2 BC). Therefore, high-dose Astragalus extract was selected for treatment and efficacy evaluation.

[0052] The researchers then investigated the effects of double treatment with high-dose Astragalus extract. The mice were given high-dose Astragalus extract during heat shock and 15 hours after recovery. The effects of the double treatment were observed 24 hours after heat shock recovery. Double treatment with high-dose Astragalus extract also significantly improved liver damage induced by extreme heat ( Figure 2 DE).

[0053] Example 4: Effect of Astragalus Extract Treatment on Phosphatidic Acid Phosphohydrolase (Lipin1) in Mouse Liver;

[0054] Compared with the control group, the mRNA expression level of Lipin 1 in the liver of mice in the heat exposure group was significantly decreased. After a single high-dose Astragalus extract treatment, the mRNA expression of Lipin 1 was significantly increased ( Figure 3 A). Compared with the NC group, the protein expression level of Lipin 1 in the liver of mice in the HC group was significantly decreased; compared with the HC group, the protein expression level of Lipin 1 in the liver of mice in the HA group was significantly increased ( Figure 3 BC). The Lipin 1 gene, a core factor in lipid metabolism regulation, was first confirmed in 2001 to be significantly associated with abnormal lipid deposition. The Lipin 1 protein encoded by this gene belongs to the Lipin protein family and primarily functions as a phosphatidic acid phosphohydrolase in cells, catalyzing the dephosphorylation of phosphatidic acid to produce diacylglycerol. This reaction is the rate-limiting step in the triglyceride biosynthesis pathway and plays a decisive role in maintaining normal lipid storage. Studies have shown that when Lipin 1 expression levels are reduced, it leads to abnormal accumulation of diacylglycerol in the liver, which in turn induces lipid metabolism disorders by interfering with the insulin receptor signaling pathway, ultimately leading to pathological lipid deposition. These results suggest that a single high-dose Astragalus extract treatment can significantly enhance the expression of Lipin 1 in the liver of mice exposed to extreme heat and improve liver lipid metabolism.

[0055] Example 5: Effects of Astragalus Extract Treatment on the Intestinal Mucosal Barrier in Mice;

[0056] Compared with the control group, the serum D-Lactate, LPS and DAO levels of mice in the heat exposure group were significantly increased; compared with the heat exposure group, the serum LPS level of mice exposed to extreme high temperature after a single high dose of Astragalus extract treatment was significantly reduced, and the levels of D-Lactate and DAO showed a downward trend ( Figure 4AC). DAO is a key biomarker for assessing intestinal barrier function. Increased serum DAO levels indicate damage to intestinal mucosal epithelial cells and reflect that the structural and functional integrity of the intestinal mechanical barrier is destroyed. D-Lactate is a metabolite of intestinal commensal flora, and its increased blood concentration can be used as a specific indicator of abnormally increased intestinal mucosal permeability. LPS, as the main component of the outer membrane of Gram-negative bacteria, is effectively blocked by the intestinal barrier in a healthy state. However, when the intestinal mucosal barrier function is impaired, LPS can translocate into the blood circulation, triggering endotoxemia and systemic inflammatory response, becoming an important molecular marker for assessing intestinal barrier integrity. The above results indicate that a single high-dose Astragalus extract treatment enhances the intestinal mucosal barrier function of mice exposed to extreme high temperatures to a certain extent.

Claims

1. Application of Astragalus extract in the preparation of drugs for treating heatstroke, drugs for treating heatstroke combined with liver damage and / or drugs for enhancing the intestinal mucosal barrier function.

2. The use according to claim 1, characterized in that The astragalus extract is used as one of the main active ingredients or the only active ingredient of the medicine.

3. The use according to claim 1, characterized in that The drug further comprises a pharmaceutically acceptable carrier.

4. The use according to claim 1, wherein The heat stroke is severe heat stroke.

5. The use according to claim 1, characterized in that The heat stroke combined with liver damage is heat stroke combined with acute liver damage.

6. The use according to claim 1, wherein The astragalus extract is prepared by the following method: 1) Select raw Astragalus root with normal color and no moldy odor, wash it, dry it and set aside; 2) Place the raw Astragalus root from step 1) in an extraction tank, add water at a mass ratio of 1:6, open the extraction tank, heat and boil at a temperature of 120-130°C, and maintain the boiling temperature for 60 minutes. Pour the boiled raw material juice into a double-effect concentrator for concentration and sterilization; 3) homogenizing the concentrated and sterilized liquid obtained in step 2) in a homogenizer at a pressure of 20-25 MPa; 4) The extract obtained by homogenization in step 3) is placed in a spray drying tower for spray drying, with the inlet air temperature controlled between 150-190°C and the outlet air temperature controlled between 80-90°C. The feed rate is adjusted according to the outlet air temperature to obtain the astragalus extract.

7. A drug for treating heatstroke, heatstroke combined with liver damage and / or enhancing intestinal mucosal barrier function, characterized in that: One of the main active ingredients or the only active ingredient of this medicine is astragalus extract.

8. The drug for treating heat stroke, heat stroke combined with liver damage and / or enhancing intestinal mucosal barrier function according to claim 7, characterized in that: The drug further comprises a pharmaceutically acceptable carrier.

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