Application of tea source element in anti-hypoxia

Abstract

The invention provides an application of a tea source element in anti-hypoxia. The tea source element can be used for protecting multiple organs: improving functional impairment of brain, liver and lung under hypoxia stress at the same time; the composition has a multi-mechanism synergistic effect: the protective effect is achieved by adjusting energy metabolism, relieving oxidative stress, inhibiting inflammatory response and the like; the tea source element is derived from a natural product, is high in safety and is suitable for long-term use.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of tea extracts in anti-hypoxia. Background Technology

[0002] Hypoxia refers to a pathological process in which insufficient oxygen supply or impaired utilization by tissues leads to abnormal changes in the body's metabolism, function, and morphological structure. Vital organs such as the brain and heart are extremely sensitive to hypoxia; severe hypoxia can rapidly cause irreversible damage and even death. Furthermore, hypoxemia caused by a significant decrease in arterial blood oxygen content can further exacerbate tissue energy metabolism disorders, leading to multiple organ failure.

[0003] Normal bodily functions are highly dependent on oxygen; from cellular metabolism to organ function, oxygen supply is crucial. Hypoxia is essentially an adaptive or pathological response of cells to low oxygen levels. In acute severe or toxic hypoxia, mitochondrial energy metabolism disorders dominate, manifested as reduced ATP production and impaired ion pump function (such as Na+ / K+-ATPase, Ca2+, and Ca2+). 2 + / Mg 2 Decreased ATPase activity leads to cell edema, acidosis, and lysosomal membrane rupture, ultimately causing cell necrosis. Chronic mild hypoxia mainly activates compensatory regulation mediated by oxygen sensors (such as HIF-1α), but long-term hypoxia can still cause tissue damage.

[0004] High-altitude environments are characterized by low pressure and low oxygen levels. People who rapidly ascend to high altitudes often experience acute mountain sickness (such as high-altitude cerebral edema and pulmonary edema) due to hypoxia, seriously threatening their lives and health. Currently, the development of anti-hypoxia drugs faces several major challenges. First, the mechanisms of hypoxia are complex, involving multiple pathways such as energy metabolism disorders, oxidative stress, and inflammatory responses. Existing drugs mostly target a single mechanism, resulting in limited efficacy. Second, while some anti-hypoxia drugs have certain therapeutic effects, long-term use may cause adverse reactions. For example, acetazolamide commonly causes side effects such as electrolyte imbalance, sensory abnormalities, and metabolic acidosis, limiting its long-term application. Third, natural drugs are costly and have limited sources. Some naturally extracted anti-hypoxia components are expensive and difficult to popularize due to complex extraction processes and scarce resources. For example, rhodioloside mainly relies on extraction from wild rhodiola rosea in high-altitude areas, resulting in scarce raw materials, high cultivation costs, and high market prices. Ideal anti-hypoxia drugs should have multi-target synergistic effects, such as simultaneously regulating ATPase, anti-inflammatory, and antioxidant effects; high safety and few side effects; and controllable cost and stable source.

[0005] Based on the above issues, it is of great significance to develop novel, highly effective, and safe anti-hypoxia drugs. Summary of the Invention

[0006] The present invention aims to at least partially solve one of the technical problems in the above-mentioned technology, namely, to provide the application of tea extract in anti-hypoxia, which can be used to prevent and treat hypoxic diseases, thereby increasing its application scope.

[0007] Therefore, in a first aspect of the present invention, the application of tea catechins in the preparation of anti-hypoxia drugs is proposed.

[0008] According to the application of the present invention, tea extract can protect multiple organs: simultaneously improve the functional damage of the brain, liver, and lungs under hypoxic stress; it has a multi-mechanism synergistic effect: exerting protective effects through multiple pathways such as regulating energy metabolism, reducing oxidative stress, and inhibiting inflammatory responses; and it is natural and safe: tea extract is derived from natural products, has high safety, and is suitable for long-term use.

[0009] Optionally, the purity of the tea deriver is 50%.

[0010] Optionally, the tea extract can prolong the survival time of the test subjects under hypoxic conditions.

[0011] In a second aspect of the invention, the use of tea catechins in the preparation of medicaments for the prevention and treatment of hypoxia-related diseases is provided.

[0012] Optionally, the hypoxia-related diseases include energy metabolism disorders, ATPase system dysfunction, inflammatory responses, brain tissue damage, and abnormal lung function.

[0013] The tea-derived components can reduce serum CK and LDH levels under hypoxic conditions; enhance the body's tolerance to hypoxia; and significantly prolong the survival time of mice under normobaric hypoxic conditions. The tea-derived components can also increase Na+ / K+-ATPase and Ca2+ levels in brain tissue under hypoxic conditions. 2 + / Mg 2 It enhances the activity of +-ATPase, maintains ion homeostasis and energy supply in brain tissue, and prevents brain damage caused by hypoxia, thus exhibiting significant neuroprotective effects.

[0014] In a third aspect of the invention, the use of the aforementioned catechins in the preparation of a medicament for treating hypoxic tissue damage is provided, wherein the catechins can improve brain tissue damage caused by hypoxia. Specifically, the catechins can improve pathological damage to lung tissue caused by acute hypoxic stress and protect lung function.

[0015] In a fourth aspect of the invention, the use of the above-mentioned tea extract in the preparation of a drug for alleviating liver injury caused by acute hypoxic stress is provided. Specifically, the tea extract can reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, thereby alleviating liver injury caused by acute hypoxic stress.

[0016] In a fifth aspect of the invention, an anti-hypoxia drug is provided, which is prepared by adding excipients to theabrownin as an active ingredient.

[0017] According to embodiments of the present invention, the anti-hypoxia drug contains catechins that can prolong hypoxic survival time: catechins can significantly prolong the survival time of mice under normobaric hypoxia, increase serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels, and enhance the body's tolerance to hypoxia; they can protect liver function: catechins can reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, alleviating liver damage caused by acute hypoxic stress; they have anti-inflammatory effects: catechins can significantly reduce the levels of inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), alleviating the inflammatory response caused by hypoxia; and they can improve brain tissue energy metabolism: catechins can improve Na+,K+-ATPase and Ca2+ levels. 2 +,Mg 2 The activity of +-ATPase maintains the ion homeostasis and energy supply of brain tissue, prevents brain damage caused by hypoxia, and has a significant neuroprotective effect; it can also reduce lung tissue damage: tea extract can improve the pathological damage of lung tissue caused by acute hypoxic stress and protect lung function.

[0018] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0019] Figure 1 The study investigated the effects of tea-derived compounds on the serum levels of CK, LDH, ALT, and AST in mice, comparing them with the normal control group. ## P<0.01; compared with the hypoxia model group, * P<0.05, ** P<0.01;

[0020] Figure 2 Tea-derived substances and their effects on Na+ in mouse brain tissue + K + -ATPase, Ca 2 +Mg 2 The effect of +-ATPase levels, in comparison with the normal control group. ## P<0.05; compared with the hypoxia model group, * P<0.05, ** P<0.01, *** P<0.001;

[0021] Figure 3 The study investigated the effects of tea extracts on serum levels of inflammatory factors IL-6 and TNF-α in mice, comparing them with the normal control group. ##P < 0.01; compared with the hypoxia model group, * P < 0.05, ** P < 0.01, *** P < 0.001;

[0022] Figure 4 are the results of H&E staining of mouse lung tissue sections;

[0023] Figure 5 are the results of H&E staining of mouse brain tissue sections. Detailed implementation manners

[0024] The technical solutions of the present invention are illustrated by specific specific examples below. It should be understood that one or more method steps mentioned in the present invention do not exclude the existence of other method steps before and after the combined steps or the insertion of other method steps between these clearly mentioned steps; it should also be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. Moreover, unless otherwise stated, the numbers of each method step are only convenient tools for identifying each method step, rather than limiting the arrangement order of each method step or the scope in which the present invention can be implemented. The change or adjustment of their relative relationship, without substantial change in technical content, should also be regarded as the scope in which the present invention can be implemented.

[0025] To better understand the above technical solutions, the exemplary embodiments of the present invention are described in more detail below. Although the exemplary embodiments of the present invention are shown, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present invention and to fully convey the scope of the present invention to those skilled in the art.

[0026] The test materials used in the present invention are all ordinary commercially available products, which can be purchased on the market or can be prepared by known methods.

[0027] The tea source element was purchased from Xiamen Hepeng Biotechnology Co., Ltd., with a purity of 50% (tea brown pigment: dextrin = 1:1).

[0028] Salidroside was purchased from Shanghai Macklin Biochemical Co., Ltd., with a purity of ≥98%.

[0029] The experimental animals were SPF-grade ICR mice, produced by Jinan Pengyue Experimental Animal Breeding Co., Ltd., with a license number: SCXK (Lu) 20220006, and were quality tested on May 23, 2024.

[0030] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are merely descriptive and do not limit the present invention in any way.

[0031] Example 1

[0032] This study simulated hypoxia in mice under normal pressure and closed environment, using rhodioloside, a drug used in clinical treatment of hypoxia, as a positive control. By observing the effect of tea extract on the survival time of hypoxic mice, the study preliminarily evaluated the antihyperxic effect of tea extract on mice under hypoxic environment at the overall mouse level, and investigated the antihyperxic effect of tea extract.

[0033] 1) Animal grouping and treatment

[0034] Sixty healthy male ICR mice were acclimatized for 7 days and then randomly divided into 6 groups of 10 mice each: a blank control group (Control group), an acute hypoxia model group (Model group), a rhodioloside positive control group (Sal group), a low-dose catechin group (TB-L group, 40 mg / kg), a medium-dose catechin group (TB-M group, 400 mg / kg), and a high-dose catechin group (TB-H group, 4000 mg / kg). Mice in the blank and model groups were administered 200 μL of physiological saline by gavage daily, while the positive control group was administered 200 μL of rhodioloside solution (25 mg / kg) by gavage. The low, medium, and high-dose catechin groups were administered 200 μL of catechin solution by gavage. This gavage treatment continued for 28 days, during which time mice had free access to food and water, and their body weight was measured every 3 days.

[0035] Modeling method: Mice were placed in a 250mL wide-mouth bottle containing 5g of soda lime. The bottle mouth was coated with Vaseline to ensure airtightness. The cap was then tightly closed, and timing began. The mortality criteria for the mice were the same as in the hypoxic mixed gas hypoxia experiment. The time from the moment the bottle was sealed until death was defined as the hypoxia survival time. Death criteria: The mouse twitched and struggled in its hind and hind toes, then suddenly became limp, with no nasal flaring and cessation of breathing.

[0036] 2) Indicator Measurement

[0037] Serum levels of CK, LDH, ALT, and AST were measured; brain tissue ATPase was measured; lung tissue inflammatory factors (IL-6, TNF-α) were measured; and brain and lung tissue sections were stained with H&E.

[0038] 3) Results Analysis

[0039] Analysis of atmospheric pressure closed hypoxia experiment:

[0040] Table 1. Effects of tea extract on survival time of normobaric closed-loop hypoxic mice (x±SD, n=6)

[0041]

[0042] Note: Compared with the model group: *P<0.05, **P<0.01, ***P<0.001

[0043] Six mice from each group underwent a normobaric closed-loop hypoxia experiment, and hypoxia models were established for each mouse. Survival time under hypoxic conditions was analyzed by detecting anti-hypoxia time, as shown in Table 1. Compared with the model group without any drug treatment, mice treated with rhodioloside showed a significant increase in survival time under hypoxia, with a survival time increase of 45.91% (P < 0.001). Mice treated with catechins also showed a significant increase in survival time under hypoxia, with corresponding survival rate increases of 29.21% (P < 0.05), 35.96% (P < 0.01), and 23.89% (P < 0.05), respectively. This indicates that catechins can prolong the anti-hypoxia survival time of mice under hypoxic conditions and possess a certain anti-hypoxia ability.

[0044] Analysis of mouse serum CK, LDH, ALT, and AST levels:

[0045] The CK, LDH, ALT, and AST detection kits provided by Nanjing Jiancheng Bioengineering Research Institute were used to quantitatively determine the levels of these substances in mouse serum. Figure 1 As shown, acute hypoxic stress significantly affected energy metabolism and liver function in mice. Figure 1 As shown in Figures A and B, serum CK and LDH levels in the model group mice were significantly higher than those in the blank control group (P < 0.01), confirming that hypoxia successfully induced metabolic disorders and tissue damage. Notably, the Sal group and different doses of tea extract intervention groups (low, medium, and high) effectively reduced serum LDH levels (P < 0.05 or P < 0.01), while CK levels in all dose groups also showed a decreasing trend. This suggests that tea extract may enhance the body's hypoxia tolerance by regulating energy metabolism and alleviating oxidative stress.

[0046] In terms of liver function assessment, such as Figure 1 As shown in C and 1D, serum ALT and AST levels in the model group mice were significantly higher than those in the normal group (P < 0.01), indicating that hypoxia indeed caused liver damage. All doses of tea extract (low, medium, and high) significantly reduced serum ALT levels (P < 0.05 or P < 0.01). Regarding AST levels, the Sal group and the low and medium dose tea extract groups also showed significant improvement (P < 0.05). These findings strongly demonstrate that tea extract can effectively protect the liver from hypoxic damage, and its mechanism of action may be related to inhibiting transaminase release, thereby enhancing the body's hypoxia tolerance.

[0047] ATPase analysis of mouse brain tissue:

[0048] Na produced by Nanjing Jiancheng Biotechnology Research Institute + K + -ATPase and Ca 2+ Mg2+ - An ATPase assay kit is used to quantitatively determine the activity of ATPase in mouse brain tissue. For example... Figure 2 As shown, this reveals the regulatory effect of tea glycosides on ATPase activity in the brain tissue of hypoxic mice. Figure 2 As shown in A and 2B, compared with the normal control group, the Na+ level in the acute hypoxia model group was significantly lower. + K + -ATPase and Ca 2+ Mg 2+ -ATPase activity was significantly decreased in both groups (P < 0.05). After tea-derived protein intervention, the low- and medium-dose groups significantly increased Na+. + K + -ATPase activity (P < 0.05), while the medium and high dose groups significantly increased Ca2+ activity (P < 0.05). 2+ Mg 2+ -ATPase activity (P < 0.05). This indicates that tea extract can maintain brain tissue ion balance and energy metabolism by enhancing the function of the ATPase system, thereby effectively preventing nerve damage caused by hypoxia and showing a significant neuroprotective effect.

[0049] Analysis of lung tissue inflammatory factors IL-6 and TNF-α:

[0050] The levels of inflammatory factors in the lung tissue of experimental mice were quantitatively analyzed using an IL-6 and TNF-α ELISA kit manufactured by Wuhan Beinlai Biotechnology Co., Ltd. Experimental data showed that hypoxic stress significantly promoted the release of inflammatory factors. Figure 3 As shown, serum IL-6 and TNF-α levels in the model group mice were significantly higher than those in the normal control group (P < 0.01), demonstrating that hypoxia successfully induced an inflammatory response. After drug intervention, the Sal group and the medium and high dose groups of tea extract significantly reduced TNF-α levels (P < 0.05 or P < 0.01), and the low dose group also showed a decreasing trend. For IL-6 levels, the Sal group and all dose groups of tea extract (low, medium, and high) showed significant inhibitory effects (P < 0.05 or P < 0.01). These results fully demonstrate that tea extract has significant anti-inflammatory effects, can effectively reduce the inflammatory response caused by hypoxia, and thus enhance the body's tolerance to hypoxia.

[0051] Analysis of H&E stained sections of lung and brain tissue:

[0052] Mouse brain and lung tissues were cleaned with physiological saline and then fixed in a tissue fixative to maintain their original morphology and structure. The tissues were then embedded in paraffin and allowed to solidify into a waxy, translucent mass. The embedded tissues were sectioned, stained, and scanned under a microscope. This embedding and sectioning work was performed by Wuhan Saiweier Biotechnology Co., Ltd. The pathological observation results of the lung tissue are as follows: Figure 4 As shown in the figure, the lung tissue of the normal control group mice exhibited typical physiological structural features, including intact alveolar structure, clear alveolar cavities, and uniformly distributed thin alveolar septa, without any signs of inflammation or edema. In contrast, the acute hypoxia model group showed obvious pathological damage features: disordered lung tissue structure, swollen and deformed alveolar cavities, significantly thickened alveolar septa, and diffuse hemorrhagic edema. In addition, significant microcirculatory disturbances were observed in this group, manifested as dilation and congestion of pulmonary arterioles and capillaries, as well as microthrombus formation. The tea extract treatment group showed a dose-dependent protective effect, with all dose groups effectively improving the above pathological changes, including restoring the integrity of alveolar wall structure, reducing hemorrhage points, improving vascular morphology, and inhibiting inflammatory cell infiltration. Notably, the protective effect of the high-dose tea extract group was significantly better than that of the positive control Sal group.

[0053] Brain tissue pathological analysis results as follows Figure 5 As shown, the normal control group's brain tissue exhibited typical neuronal arrangement characteristics, with regular nuclei morphology and a clear boundary between the cytoplasm and nuclear membrane. The model group, however, showed obvious characteristics of hypoxic injury, including a decrease in the number of neurons, nuclear pyknosis, widening of intercellular spaces, and vacuolation. The experimental results showed that all doses of tea extract significantly improved these pathological changes, with the high-dose group showing a protective effect comparable to the positive control Sal group. This indicates that tea extract can exert a neuroprotective effect through mechanisms such as reducing cellular edema and inhibiting neuronal apoptosis.

[0054] In summary, embodiments of the present invention demonstrate that tea extract can prolong hypoxic survival time: tea extract can significantly prolong the survival time of mice under normobaric hypoxia, increase serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels, and enhance the body's tolerance to hypoxia; it can protect liver function: tea extract can reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, alleviating liver damage caused by acute hypoxic stress; it has anti-inflammatory effects: tea extract can significantly reduce the levels of inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), alleviating the inflammatory response caused by hypoxia; and it can improve brain tissue energy metabolism: tea extract can improve brain tissue energy metabolism by increasing Na+,K+-ATPase and Ca2+. 2 +,Mg 2 The activity of +-ATPase maintains the ion homeostasis and energy supply of brain tissue, prevents brain damage caused by hypoxia, and has a significant neuroprotective effect; it can also reduce lung tissue damage: tea extract can improve the pathological damage of lung tissue caused by acute hypoxic stress and protect lung function.

[0055] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. The illustrative expressions of the above terms in this specification should not be construed as necessarily referring to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. In addition, those skilled in the art can combine and integrate the different embodiments or examples described in this specification.

[0056] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. Use of theaflavins in the preparation of an anti-hypoxia drug.

2. Use according to claim 1, wherein The purity of the theaflavins is 50%.

3. The use according to claim 1, wherein The theaflavins can prolong the survival time of a subject under hypoxic conditions.

4. Use of theaflavins in the preparation of a drug for preventing and treating hypoxia-related diseases.

5. The use according to claim 4, wherein the compound is ###0002### The hypoxia-related diseases include energy metabolism disorders, ATP enzyme system dysfunction, inflammatory reactions, brain tissue damage, and lung function abnormalities.

6. The use according to claim 4, wherein the compound is ###0002### The theaflavins can reduce the levels of creatine kinase (CK) and lactate dehydrogenase (LDH) in serum under hypoxic conditions.

7. The use according to claim 4, wherein the compound is ###0002### The tea source can improve the activity of Na+ / K+-ATPase and Ca 2 + / Mg 2 +-ATPase in brain tissue under hypoxic conditions.

8. Use of the theasinensin in the preparation of a medicament for the treatment of hypoxic tissue damage, characterized in that, The theaflavins can improve brain tissue damage caused by hypoxia.

9. Use of theaflavins in the preparation of a drug for reducing liver damage caused by acute hypoxic stress.

10. An anti-hypoxic agent, characterized in that it is a compound of general formula (I) or its pharmaceutically acceptable salt. The drug is prepared by adding excipients to theaflavins as an active ingredient.

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