Application of ATTM in preparation of medicine for preventing or / and treating heat stroke

Abstract

The invention belongs to the field of medicines, and discloses application of ATTM in preparation of a medicine for treating heat stroke. According to the application, ATTM is researched, so that the ATTM can effectively reduce the level of proinflammatory factors, improve multi-organ injury caused by thermal stimulation and remarkably reduce the death rate caused by thermal stimulation, has a treatment effect on the heat stroke when being applied to preparation of the medicine for treating the heat stroke, and can be used as a novel candidate treatment medicine for the heat stroke.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to an application of ATTM in preparing a drug for preventing and / or treating heat stroke. Background Art

[0002] Heat stroke is an acute illness caused by impaired thermoregulation due to high temperatures or certain physical, chemical, or biological factors, resulting in excess heat production and heat dissipation, leading to abnormal changes in the central nervous system. Clinically, heat stroke is characterized by extreme body temperature elevation (core temperature typically exceeding 40°C), systemic inflammation, circulatory collapse, bleeding and coagulopathy, and multiple organ dysfunction. These conditions are caused by a complex interaction between heat-related cytotoxicity, inflammation, and disseminated intravascular coagulation (DIC).

[0003] Heatstroke is common among young adults who engage in physical labor in hot and humid environments (exertional heatstroke) and among the elderly with impaired temperature regulation (classic heatstroke). Epidemiological data show that the mortality rate for severe cases, if left untreated, can exceed 60%, and 30%-40% of survivors suffer permanent neurological sequelae. Heatstroke has extremely high mortality and disability rates and is a common central nervous system injury in the summer.

[0004] Currently, there is a lack of specific therapeutic drugs for heat stroke, and the main clinical treatment options are physical cooling, organ support therapy, and drug intervention. Cooling therapy: Physical cooling such as cold water immersion and ice blankets is the core measure, but the cooling rate is directly related to the patient's prognosis, and advanced patients are often difficult to cool down effectively due to circulatory failure. Organ support therapy: Mechanical ventilation, continuous renal replacement therapy (CRRT), etc. can temporarily maintain vital signs, but cannot reverse the cellular cascade damage that has occurred. Drug intervention: Glucocorticoids, antioxidants (such as N-acetylcysteine), etc. have been tried to inhibit inflammation and oxidative stress, but the effect is not good.

[0005] However, the current treatment time window for heat stroke is narrow: the pathological progression of heat stroke worsens exponentially, and the success rate of treatment decreases significantly after the "golden 6 hours". There is an urgent need for fast-acting nerve and organ protectors; moreover, there are currently no FDA- or NMPA-approved specific drugs for heat stroke, and clinical medications are mostly based on empirical treatment and lack high-quality evidence-based medical support.

[0006] Therefore, it is of great significance to seek a new therapeutic drug to improve the treatment effect of heat stroke. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the deficiencies and defects mentioned in the above background technology and provide an application of ATTM in the preparation of drugs for preventing and / or treating heat stroke.

[0008] In order to solve the above technical problems, the technical solution proposed by the present invention is: A use of ATTM in preparing a drug for preventing and / or treating heat stroke, wherein the ATTM refers to ammonium tetrathiomolybdate.

[0009] In the above application, preferably, the drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0010] Compared with the prior art, the present invention has the following beneficial effects: This application studies ATTM, which can effectively reduce the level of pro-inflammatory factors, improve multi-organ damage caused by heat stimulation, and significantly reduce the mortality rate caused by heat stimulation. It is used in the preparation of drugs for treating heat stroke, has a therapeutic effect on heat stroke, and can be used as a new candidate preventive and therapeutic drug for heat stroke. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 is the HE staining result of the inflammatory cell infiltration degree of mouse tissues in each group; Figure 2 These are the test results of blood transaminases, creatinine, and inflammatory factors in each group of mice; Figure 3 is the mortality rate of mice in each group. DETAILED DESCRIPTION

[0012] To facilitate understanding of the present invention, the present invention will be described in more comprehensive and detailed form below in conjunction with the accompanying drawings and preferred embodiments. However, the protection scope of the present invention is not limited to the following specific embodiments.

[0013] Unless otherwise defined, all technical terms used hereinafter have the same meanings as those generally understood by those skilled in the art. The technical terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention.

[0014] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0015] In the following examples, all measured data are expressed as ± s. One-way analysis of variance (ANOVA) was used for comparisons among multiple groups, and log-rank analysis was used for mouse mortality curves. P < 0.05 was considered statistically significant. (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) Example 1: A use of ATTM in preparing a drug for preventing and / or treating heat stroke.

[0016] To demonstrate the effectiveness of ATTM in preparing drugs for preventing and / or treating heat stroke, this example conducted an efficacy study and verification on mice. The specific experimental process is as follows: 1. Materials and Reagents ATTM (CAS: HY-W076067, MCE); IL-1β, TNF-α ELISA kits (Proteintech); 2. Experimental Animals Clean-grade male C57BL / 6J mice were purchased from Changsha Slake Animal Co., Ltd.

[0017] 3. Experimental Methods 1. Prepare a heat stroke mouse model induced by thermal stimulation and observe the efficacy of ATTM in treating heat stroke Forty clean-grade C57BL / 6J mice (8 weeks old, male, weighing 20-22 g) were numbered and randomly divided into five groups (8 mice per group): control group, heat stroke model group, heat stroke + ATTM 10 mg / kg treatment group, heat stroke + ATTM 20 mg / kg treatment group, and heat stroke + ATTM 40 mg / kg treatment group. The treatment groups received intraperitoneal injections of ATTM starting 3 days before modeling and administered once daily for 3 days. The control and model groups received an equal volume of normal saline intraperitoneally. The heat stroke mouse model was established in an artificial climate chamber with a temperature of 39°C and a humidity of 60 ± 5%. Every 10 minutes, mice were connected to a Powlalr 8sp physiological recorder with a thermocouple placed in the rectum. Rectal temperature (Tr) was recorded and used as a proxy for core body temperature (Tc). Mice were removed from the climate chamber when Tr reached 43°C. The control group received no heat stimulation. During the heat stimulation, mice were deprived of food and water. After the heat stimulation ended, they resumed eating and drinking and were returned to their original housing environment. Twelve hours after the heat stimulation ended, the mice were anesthetized and blood was collected for serum, and then they were sacrificed.

[0018] The mouse chest cavity was opened and the lungs were removed. After sufficient lavage of the right ventricle, the left lung was fixed in 10% paraformaldehyde for pathological examination. The right lung was lobed and placed in a liquid nitrogen tank. The degree of lung inflammatory cell infiltration was observed by HE staining of paraffin sections. The mouse abdominal cavity was opened and the mouse liver was removed. The left lateral lobe was retained for paraffin sectioning, and the remaining liver tissue was placed in a liquid nitrogen tank. The paraffin sections were stained with HE to observe liver damage. The bilateral kidney tissue was separated and fully lavaged. The left kidney was retained for paraffin sectioning, and the renal cortex of the remaining kidney was retained and placed in a liquid nitrogen tank. The renal function of each group of mice was tested by the picric acid two-point rate method, and the paraffin sections were stained with HE to observe renal tubular damage.

[0019] Alveolitis was scored using the same criteria as Susan V. et al. to analyze the degree of lung inflammation: Grade 0 (score 0) = no alveolitis; Grade I (score 1) = mild alveolitis, characterized by mononuclear cell infiltration and thickening of the alveolar septa, but with localized lesions occupying less than 20% of the lung, and with essentially normal alveolar architecture; Grade II (score 2) = moderate alveolitis, characterized by involvement occupying approximately 20% to 50% of the lung; Grade III (score 3) = severe alveolitis, characterized by involvement occupying 50% to 75% of the lung, with occasional intraalveolar consolidation caused by mononuclear cells and hemorrhage; and Grade IV (score 4) = severe alveolitis, characterized by involvement occupying greater than 75% of the lung, with intraalveolar consolidation caused by mononuclear cells and hemorrhage. For each slide, 10 fields of view at 100x magnification were randomly selected for scoring, and the average score was used as the sample score.

[0020] Liver tissue injury scoring was based on the semi-quantitative analysis of liver pathological damage using the method used by Carlos A. et al. The scoring criteria are: Grade 0 (score 0): minimal or no evidence of injury; Grade 1 (score 1): mild injury with cytoplasmic vacuolation and focal nuclear pyknosis; Grade 2 (score 2): moderate to severe injury with extensive nuclear pyknosis, cytoplasmic eosinophilia, and loss of intercellular borders; Grade 3 (score 3): severe necrosis with loss of hepatic cords, hemorrhage, and neutrophil infiltration. Five randomly selected fields at 200x magnification were scored for each slide, and the average score was used to represent the sample.

[0021] The renal tubular injury scoring standard was based on the method used by Capasso et al. for semiquantitative analysis of renal tubular pathological damage. Ten fields of view in the corticomedullary region were randomly selected under a 200× microscope, and the area of tubular damage (tubular necrosis, cast formation, and brush border loss) was scored as follows: 0: None; 1: 1%-25%; 2: 26%-50%; 3: 51%-75%; 4: 76%-100%. The average score was used as the sample score.

[0022] The results of alveolitis, liver tissue damage, and renal tubular damage scores are shown in Table 1 .

[0023] Table 1 Scoring results of alveolitis, liver tissue damage and renal tubular damage

[0024] HE staining results Figure 1As shown, under the light microscope, the alveolar structure of the blank control group mice was basically normal, the alveolar wall was intact, there was no obvious lesion, and no inflammatory cells were distributed; the alveolar structure of the heat stroke model group mice was obviously destroyed and disordered, a large number of mixed inflammatory cells diffusely infiltrated, and some lung tissues underwent consolidation; the degree of alveolar structure disorder and inflammatory cell infiltration in the heat stroke + ATTM 10 mg / kg treatment group, heat stroke + ATTM 20 mg / kg treatment group, and heat stroke + ATTM 40 mg / kg treatment group were significantly less than those in the model group, and basically no consolidation or bleeding was observed, and the pneumonia score was significantly reduced.

[0025] The renal tubular epithelium of heat stroke model mice showed significant cell swelling, vacuolar degeneration, necrosis, tubular dilation, and exposed basement membranes. Compared with the model group, the kidneys of mice treated with heat stroke plus ATTM 10 mg / kg, heat stroke plus ATTM 20 mg / kg, and heat stroke plus ATTM 40 mg / kg showed significant alleviation of these lesions, a smaller range of damage, and significantly lower renal tubular injury pathology scores.

[0026] The livers of heat stroke model mice showed extensive loss of intercellular boundaries, loss and necrosis of hepatic cords, sinusoidal congestion, and nuclear pyknosis. Compared with the model group, mice treated with ATTM showed a significant reduction in these lesions, a smaller lesion area, and significantly lower liver pathology scores. The heat stroke combined with ATTM 10mg / kg treatment group showed no significant reduction in liver pathology, while the heat stroke combined with ATTM 20mg / kg group showed moderate to effective treatment. The heat stroke combined with ATTM 40mg / kg group showed the best results.

[0027] The results of blood transaminase, creatinine and inflammatory factors in each group of mice are shown in Figure 2 As shown in the results, compared with the control group, the average values of blood transaminase, blood creatinine, and blood inflammatory factors (IL-1β, TNF-α) in the heat stroke model group were significantly higher than those in the normal group (p < 0.001); after treatment with ATTM 10 mg / kg, 20 mg / kg, and 40 mg / kg, the blood inflammatory factors and blood creatinine of the mice were significantly decreased, and the differences were statistically significant; after treatment with ATTM 20 mg / kg and 40 mg / kg, the blood transaminase of the mice was significantly decreased, and the differences were statistically significant, but the blood transaminase did not decrease significantly after treatment with ATTM 10 mg / kg.

[0028] 2. Prepare a heat stroke mouse model induced by thermal stimulation and observe the efficacy of ATTM on the mortality rate of heat stroke mice Based on the previously identified optimal therapeutic dose of 40 mg / kg, this dose was used for mortality efficacy testing. Twenty-eight clean-grade C57BL / 6J mice (8-week-old, male, weighing 20-22 g) were randomly divided into two groups: a heat stroke model group and a heat stroke + ATTM 40 mg / kg treatment group. The mice in each group received the same treatment as above. Survival was observed and recorded daily for three consecutive days after model establishment.

[0029] The mortality rate of mice in each group is shown in Figure 3 As shown, the heat stroke model group experienced a large number of deaths 24 hours after the end of the heat stimulation, with a mortality rate exceeding 50%. The mortality rate reached 75% after 48 hours, and no deaths were observed thereafter. Treatment with 40 mg / kg of ATTM significantly improved the mortality rate of heat stroke mice, with a mortality rate of 25% after 24 hours, and no deaths were observed thereafter.

[0030] In summary, ATTM can effectively reduce the levels of proinflammatory factors, improve multi-organ damage caused by heat stimulation, and significantly reduce the mortality rate caused by heat stimulation. It can be used as a new candidate preventive and / or therapeutic drug for heat stroke.

[0031] The above is only a preferred embodiment of the present invention, and the scope of protection of the present invention is not limited to the above embodiment. For those skilled in the art, improvements and modifications obtained without departing from the technical concept of the present invention should also be considered as the scope of protection of the present invention.

Claims

1. Use of ATTM in the preparation of a drug for preventing and / or treating heat stroke.

2. The use according to claim 1, characterized in that The drug further comprises a pharmaceutically acceptable carrier and / or excipient.

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