Application of epigallocatechin gallate in preparation of medicine for preventing and treating high-temperature heatstroke
The drug prepared by intraperitoneal injection of EGCG inhibits cell death caused by heatstroke, solving the problem of prevention and treatment of heatstroke, improving survival rate and reducing tissue damage, and has high safety and few side effects.
Patent Information
- Application Number
- CN202511932479.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-19
- Publication Date
- 2026-01-23
AI Technical Summary
Current technologies have not effectively solved the prevention and treatment of heatstroke, especially the problem of cell death caused by excessive heat storage, which leads to high mortality and tissue damage.
Epimarginal catechin gallate (EGCG) is used as the drug component and administered via intraperitoneal injection at a dose of 0-25 mg/kg. It is used to prepare drugs for the prevention and treatment of heatstroke and to inhibit cell death and tissue damage.
EGCG significantly improved the survival rate of mice, reduced tissue damage, decreased mortality, and had high safety and few side effects.
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Figure CN121370871A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of medicine, in particular to the application of EGCG in the preparation of drugs for preventing and treating heatstroke. BACKGROUND
[0002] Heatstroke (HS) is a severe and fatal disease caused by heat damage factors directly due to long-term exposure of the body to high temperature environment. The clinical features of the disease are: the core temperature of the body exceeds 40℃ and the dysfunction of central nervous system, and the mortality rate can reach more than 60%. Due to the influence of global warming and population aging, if not effectively intervened, the number of deaths caused by heat wave will increase year by year, which will challenge the emergency response capability of the medical system.
[0003] The clinical treatment of heatstroke is divided into ordinary treatment (such as supplementing electrolyte), physical treatment (such as cooling) and drug treatment. At present, researchers have proposed many new treatment methods for different heatstroke pathogenesis, including inflammatory response, oxidative stress, coagulation dysfunction and multiple organ dysfunction, but have not been verified in clinical trials, so there is an urgent need to develop new drugs for the prevention and treatment of heatstroke to reduce the incidence and mortality of heatstroke. The potential pathogenesis of heatstroke is that the balance between heat production and heat dissipation is destroyed, resulting in excessive heat storage. Cell death directly caused by high heat is an important reason for the aggravation of symptoms in heatstroke patients. Studies have shown that the release of cytochrome c and the activation of Caspase-3 and Caspase-9 in the spleen, intestine and lung of heatstroke model of baboons and hematopoietic cells increase, thereby causing extensive apoptosis, suggesting that inhibition of cell death is a candidate strategy for the prevention and treatment of heatstroke.
[0004] Epigallocatechin gallate (EGCG) is one of the most abundant catechins in green tea, which is a typical polyphenolic flavonoid compound with 8 free hydroxyl groups. We found in animal experiments that intraperitoneal injection of EGCG can alleviate the tissue damage of various organs of heatstroke mice and improve the survival rate of mice. It is particularly worth noting that EGCG inhibits the activation of Caspase-3 and GSDME in bone marrow macrophages derived from heatstroke mice, suggesting that EGCG alleviates the death of mice caused by heatstroke by inhibiting cell death. SUMMARY
[0005] In view of the above, the present application preliminarily studies the application of EGCG in the preparation of drugs for preventing and treating heatstroke. In order to achieve the above technical effects, the present application is realized by the following technical means:
[0006] The present application first discloses the application of epigallocatechin gallate in the preparation of drugs for preventing and treating heatstroke, wherein:
[0007] The molecular formula of epigallocatechin gallate is C 12 H 18 O 11 The molecular weight is 458.37, and the chemical structural formula is as follows:
[0008] .
[0009] Further, the medicine also includes a pharmaceutically acceptable carrier.
[0010] Further, the administration route of the medicine includes intraperitoneal injection.
[0011] Further, at the time of administration, epigallocatechin gallate is a solution preparation (purchased from Sigma Company, USA, CAS number: 989-51-5) prepared in the form of a dry powder.
[0012] Further, the dosage used in intraperitoneal injection is 0-25 mg / kg.
[0013] The application also discloses a medicine for preventing and treating heat stroke, which comprises:
[0014] Epigallocatechin gallate and a pharmaceutically acceptable carrier, wherein:
[0015] The molecular formula of epigallocatechin gallate is C 12 H 18 O 11 The chemical structural formula is as follows:
[0016] .
[0017] The application has the following beneficial effects:
[0018] (1) EGCG can intervene in the pathophysiological process of heat stroke in mice by inhibiting heat-induced cell death. In a mouse heat stroke model, EGCG pretreatment can effectively inhibit heat-induced cell death, significantly improve heat-induced tissue damage in mice, and improve the survival rate of mice. Therefore, the application of EGCG in the prevention and treatment of heat stroke can effectively reduce heat-induced tissue damage and reduce the mortality rate.
[0019] (2) EGCG is a natural compound with high safety and good tolerance, and has smaller side effects and a wider application range than traditional medicines. BRIEF DESCRIPTION OF DRAWINGS
[0020] Figure 1 A is a schematic diagram of the experimental process of mouse heat stroke modeling and the prevention and treatment of heat stroke in mice by EGCG; Figure 1B is that the mice are injected with EGCG intraperitoneally, and then placed in 25℃ or 39℃ environment, and the survival curves of the control group (25℃ and 25℃+EGCG) and the experimental group (39℃ and 39℃+EGCG) mice are observed (n=15); Figure 1 C is that the mice are injected with EGCG intraperitoneally, and then placed in 25℃ or 39℃ environment, and the tissue damage of the small intestine, liver and lung of the control group (25℃ and 25℃+EGCG) and the experimental group (39℃ and 39℃+EGCG) mice is observed and analyzed by H&E staining, and the scale is 200 μm; Figure 1 D is that the mice are injected with EGCG intraperitoneally, and then placed in 25℃ or 39℃ environment, and the Caspase-3 and GSDME of the bone marrow macrophages derived from the control group (25℃ and 25℃+EGCG) and the experimental group (39℃ and 39℃+EGCG) mice are detected by WB. DETAILED DESCRIPTION
[0021] The preferred embodiments of the present application are described in detail below to make the advantages and features of the present application more easily understood by those skilled in the art, so as to make the protection scope of the present application more clearly defined.
[0022] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained commercially unless otherwise specified.
[0023] In order to make the purpose, technical scheme and advantages of the embodiments of the present application more clear, the technical scheme in the embodiments of the present application will be clearly and completely described below. The following examples are only used to illustrate the present application, and should not be regarded as limiting the scope of the present application. If the specific conditions are not specified in the examples, the conventional conditions or the conditions recommended by the manufacturer are used. If the reagents or instruments used are not specified by the manufacturer, they are all conventional products that can be purchased on the market.
[0024] Example 1
[0025] In this embodiment, a mouse heatstroke model is used to verify the effect of EGCG on the survival rate of heatstroke mice, and the specific operation method is as follows:
[0026] (1) Experimental materials:
[0027] SPF female C57BL / 6J mice (6-8 weeks, purchased from Chengdu Enswell Biological Technology Co., Ltd., body weight 18±0.5 g), 1 mL syringe, EGCG, sterile water, small animal incubator.
[0028] (2) Experimental method and specific steps:
[0029] Before the experiment, the temperature of the small animal incubator was adjusted to 25℃, and the mice were placed in the small animal incubator for 24 h to adapt to the environmental noise. The mice were intraperitoneally injected with sterile water or 25 mg / kg EGCG, and at the same time, fasting and water deprivation were performed, and the small animal incubator was adjusted to 39℃, and after 3 h, the mice were placed in a small animal incubator with a relative temperature of 39℃±0.5℃ (AT: Ambient temperature) and a relative humidity of 60%±5% (AH: Ambient humidity) for 2 h to establish a heat stroke model. After 3 h of heat stress, intraperitoneal injection of sterile water or 25 mg / kg EGCG was performed, and the mice were placed in a 25℃ environment with free access to food and water. This experimental process lasted for 6 days. On the 7th day, the mice were intraperitoneally injected with sterile water or 25 mg / kg EGCG before 3 h of heat stress, and at the same time, fasting and water deprivation were performed. Heat stress lasted for 8 h, during which the death of the mice was observed and recorded. The control group was operated in the same way but not exposed to heat stress. Figure 1 A).
[0030] (3) Experimental results:
[0031] In the control group experiment, EGCG treatment had no significant effect on the survival of mice. In the heat stress-induced heat stroke model of mice, EGCG treatment significantly reduced the mortality of mice Figure 1 B, ***p<0.01).
[0032] Example 2
[0033] This example verifies the effect of EGCG pretreatment on tissue damage in heat stroke mice through animal experiments, and the specific operation method is as follows:
[0034] (1) Experimental materials:
[0035] SPF female C57BL / 6J mice (6-8 weeks, purchased from Chengdu Enswell Biological Technology Co., Ltd., body weight 18±0.5 g), 1 mL syringe, EGCG, sterile water, small animal incubator, 4% paraformaldehyde, dimethylbenzene, anhydrous ethanol, glass jar, oven, cover glass, glass slide, paraffin, embedding box, embedding mold, tissue embedding machine, cold table, microtome, H&E staining kit, neutral gum.
[0036] (2) Experimental methods and specific steps:
[0037] Before the experiment, the temperature of the small animal incubator was adjusted to 25℃, and the mice were placed in the small animal incubator for 24 h to adapt to the environmental noise. The mice were intraperitoneally injected with sterile water or 25 mg / kg EGCG, and at the same time, they were deprived of food and water, and the small animal incubator was adjusted to 39℃. After 3 h, the mice were placed in a small animal incubator at 39℃±0.5℃ and a relative humidity of 60%±5% for 2 h to establish a heatstroke model. After 3 h of heat stress, the mice were intraperitoneally injected with sterile water or 25 mg / kg EGCG and placed in a 25℃ environment with free access to food and water. This experiment lasted for 7 days. On the 7th day, the mice were sacrificed after 2 h of heat stress, and the small intestine, liver, and lung were completely removed and fixed with 4% paraformaldehyde at room temperature for 48 h. The control group was operated on in the same way but was not exposed to heat stress.
[0038] The fixed tissues were placed in embedding boxes and labeled, dehydrated in different concentration gradients of ethanol, and then subjected to embedding, slicing, and baking steps. Finally, they were stained with H&E staining reagent, and the data were recorded by scanning the slides with a slide scanner.
[0039] (3) Experimental results:
[0040] Under heat stress, the mouse small intestine epithelium was severely damaged, the liver cells had abnormal morphology, the nucleus was shrunk, and there was inflammatory cell infiltration in the lung. When EGCG was used for prevention and treatment, the mouse small intestine epithelium was intact, the degree of liver damage was significantly alleviated, and the inflammatory cell infiltration in the lung was significantly reduced (Fig. C), indicating that EGCG has a protective effect on tissue damage caused by high temperature. Figure 1
[0041] Example 3
[0042] This example verifies the effect of EGCG pretreatment on the death of mouse bone marrow-derived macrophages through animal experiments. The specific operation method is as follows:
[0043] (1) Experimental materials:
[0044] SPF level female C57BL / 6J mice (6-8 weeks, purchased from Chengdu Enswell Biological Technology Co., Ltd., body weight 18±0.5 g), 1 mL syringe, EGCG, sterile water, small animal incubator, 70 μM filter, 5 mL syringe, red blood cell lysis solution, PBS, Lysis buffer, protease inhibitor, 6× loading buffer, Running buffer, Transfer buffer, PVDF membrane, skimmed milk powder, TBST, anti-mouse GSDME antibody, anti-mouse Caspase-3 antibody, and anti-mouse β-actin antibody.
[0045] (2) Experimental methods and specific steps:
[0046] Before the experiment, the temperature of the small animal incubator was adjusted to 25℃, and the mice were placed in the small animal incubator for 24 h to adapt to the environmental noise. The mice were intraperitoneally injected with sterile water or 25 mg / kg EGCG, and at the same time, the mice were fasted and deprived of water, and the small animal incubator was adjusted to 39℃, and after 3 h, the mice were placed in a small animal incubator at 39℃±0.5℃, relative humidity 60%±5% for 2 h to establish a heatstroke model. After 3 h of heat stress, the mice were intraperitoneally injected with sterile water or 25 mg / kg EGCG, and placed in a 25℃ environment, and allowed to eat and drink freely. The experiment lasted for 7 days. On the 7th day, the mice were sacrificed after 2 h of heat stress, and the mouse bone marrow cells were removed, the red blood cells were removed, and the cells were lysed on ice for 30 min with Lysis buffer containing protease inhibitors. After centrifugation at 12000 rpm and 4℃ for 10 min, the supernatant was added to 6x loading buffer and boiled at 100℃ for 10 min.
[0047] WB experimental procedure: 20 μL of sample was loaded and electrophoresed at 200 V for 50 min. The protein was transferred to a PVDF membrane by wet transfer, and then blocked with 5% skim milk at room temperature for 1 h. The corresponding antibody was added and incubated at 4℃ overnight. The next day, TBST was washed 3 times, and the secondary antibody was incubated at room temperature for 1 h. Finally, TBST was washed 3 times, and the band image was obtained by chemiluminescence imager.
[0048] (3) Experimental results:
[0049] Under heat stress, Caspase-3 and GSDME in bone marrow-derived macrophages in mice were activated, and when EGCG was used for prevention and treatment, the activation of Caspase-3 and GSDME was significantly inhibited, indicating that EGCG can inhibit cell death caused by heat stress Figure 1 D}.
[0050] The above summary and specific embodiments are intended to demonstrate the practical application of the technical solutions provided by the present application, and should not be interpreted as limiting the scope of protection of the present application. Those skilled in the art can make various modifications, equivalent replacements or improvements within the spirit and principles of the present application. The scope of protection of the present application is subject to the appended claims.
Claims
1. Use of epigallocatechin gallate in the preparation of a medicine for preventing and treating heat stroke at high temperature, wherein: The molecular formula of epigallocatechin gallate is C 12 H 18 O 11 , and the chemical structural formula is as follows: 。 2. The use according to claim 1, wherein: the medicament further comprises a pharmaceutically acceptable carrier.
3. The use according to claim 1, wherein: the administration route of the medicament comprises intraperitoneal injection.
4. The use according to claim 3, wherein: at the time of administration, the epigallocatechin gallate is formulated as a solution of a dry powder.
5. The use according to claim 3, wherein: the intraperitoneal injection is administered at a dose of 0-25 mg / kg.
6. A medicament for preventing and treating heat stroke, comprising: epigallocatechin gallate and a pharmaceutically acceptable carrier; wherein: the medicament is administered at a dose of 0-25 mg / kg. The molecular formula of the epigallocatechin gallate is C 12 H 18 O 11 The chemical structural formula is as follows: 。