Application of naringin in preparation of medicine for preventing and treating plateau pulmonary edema

Abstract

The invention discloses an application of naringin in preparation of a medicine for preventing and treating plateau pulmonary edema. In the construction process of the mouse plateau pulmonary edema model, the naringin drug treatment group can reduce the lung coefficient caused by the mouse plateau pulmonary edema model; meanwhile, the expression of inflammatory factors TNF-alpha, IL-1beta and IL-6 in lung tissues is inhibited; mouse lung function detection finds that the lung function can be improved; lung tissue HE staining shows that the naringin can also relieve the edema exudation level of the lung tissue structure. Naringin and sodium butyrate are further combined for use, and it is found that the prevention and treatment effect on the plateau pulmonary edema is obviously better.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and provides the application of naringin in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema. Background Technology

[0002] High-altitude pulmonary edema is a severe acute mountain sickness, referring to an acute lung disease caused by humans rapidly ascending to high-altitude areas above 3,000 meters. Its core pathological changes are increased pulmonary capillary permeability and alveolar effusion, with acute pulmonary edema and inflammatory response being prominent features.

[0003] Clinical research on the prevention and treatment of high-altitude pulmonary edema is of great significance in safeguarding the health of special populations and supporting high-altitude development and national defense. On the one hand, it can support military and emergency needs, providing medical support solutions for troops stationed on high-altitude territories and border patrol units, enhancing their combat effectiveness and emergency rescue capabilities in extreme environments. On the other hand, it can protect high-altitude residents and travelers, identifying high-risk factors and preventive measures, providing health guidance for high-altitude settlers, mountaineering enthusiasts, and aid workers, and reducing the risk of illness. Furthermore, research on the prevention and treatment of high-altitude pulmonary edema can also contribute to the development of high-altitude regions, reducing the constraints of high-altitude diseases on population movement and economic construction, and providing health support for high-altitude tourism and resource development.

[0004] In recent years, with the rise of high-altitude military operations, economic development, and tourism in high-altitude areas, the prevention and treatment of high-altitude pulmonary edema has become a hot topic in high-altitude clinical medicine, seriously threatening the survival and health of those who rapidly ascend to high altitudes. However, there are currently no specific drugs or treatments that can effectively and quickly cure high-altitude pulmonary edema, and treatment options remain limited.

[0005] The mouse model of high-altitude pulmonary edema is a classic model for studying high-altitude pulmonary edema. C57BL / 6J mice were exposed to a small animal hypobaric chamber for 48 hours, simulating an altitude of 6000 meters. Early exposure to high altitudes led to DNA damage in lung tissue cells, resulting in the production of large amounts of reactive oxygen species in the cytoplasm, followed by the release of numerous inflammatory factors triggering an inflammatory response. High altitude reduces the partial pressure of inhaled oxygen, causing spasm and constriction of pulmonary arterioles. This vasoconstriction caused a sharp increase in pulmonary artery pressure, damaging the pulmonary capillary endothelial barrier. Subsequently, increased capillary permeability allowed fluid, proteins, and even red blood cells from the blood to seep into the alveoli, leading to pulmonary edema and loss of gas exchange function. Furthermore, capillary damage and pulmonary edema resulted in the release of inflammatory factors in the lungs, triggering an inflammatory response.

[0006] Naringin is a natural flavonoid compound with a core structure of a flavonoid nucleus bound to a glycosyl group. It possesses unique chemical properties and is widely found in citrus fruits, being one of the main sources of their bitterness. Reports indicate that naringin has antioxidant, anti-apoptotic, and anti-inflammatory effects; however, its role in the prevention and treatment of high-altitude pulmonary edema has not been reported. Therefore, exploring the role of naringin in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema is of significant research importance. Summary of the Invention

[0007] In view of this, the purpose of the present invention is to provide the application of naringin in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema.

[0008] To achieve the above objectives, the present invention provides the following technical solution: 1. Application of naringin in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema.

[0009] The structural formula of naringin is as follows: .

[0010] As one of the preferred technical solutions, the plateau has an altitude of over 6,000 meters.

[0011] As one of the preferred technical solutions, naringin can prevent and treat high-altitude pulmonary edema by inhibiting the expression of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissue.

[0012] As one of the preferred technical solutions, naringin can prevent and treat high-altitude pulmonary edema by improving lung function.

[0013] 2. A combination of naringin and sodium butyrate.

[0014] As one of the preferred technical solutions, the mass ratio of naringin to sodium butyrate is 4:3.

[0015] As one of the preferred technical solutions, the composition prevents and treats high-altitude pulmonary edema by inhibiting the expression of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissue.

[0016] As one of the preferred technical solutions, the composition prevents and treats high-altitude pulmonary edema by improving lung function.

[0017] 3. Application of the aforementioned composition in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema.

[0018] As one of the preferred technical solutions, the plateau has an altitude of over 6,000 meters.

[0019] 4. A drug for the prevention and treatment of high-altitude pulmonary edema, the active ingredient of which is naringin.

[0020] 5. A drug for the prevention and treatment of high-altitude pulmonary edema, comprising the aforementioned composition.

[0021] The beneficial effects of this invention are: This invention provides the application of naringin in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema. The applicant discovered that, during the construction of a mouse model of high-altitude pulmonary edema, the naringin-treated group could reduce the lung coefficient induced by the mouse model; simultaneously, it inhibited the expression of inflammatory factors TNF-α, IL-1β, and IL-6 in lung tissue; mouse lung function tests showed that it could improve lung function; and HE staining of lung tissue showed that naringin could also reduce the level of edema and exudation in lung tissue structure.

[0022] In summary, naringin can be used to prevent and treat high-altitude pulmonary edema induced by hypoxia in high-altitude low-pressure environments in mice, specifically by: reducing the lung coefficient; inhibiting the expression of inflammatory factors in lung tissue; improving lung function; and alleviating the pathological changes of high-altitude pulmonary edema in the lungs. Therefore, naringin can be considered a potential drug for the prevention and treatment of high-altitude pulmonary edema.

[0023] The applicant further combined naringin and sodium butyrate and found that the effect on the prevention and treatment of high-altitude pulmonary edema was significantly better.

[0024] Other advantages, objectives, and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination, or may be learned from practice of the invention. The objectives and other advantages of the invention can be realized and obtained through the following description. Attached Figure Description

[0025] To make the objectives, technical solutions, and advantages of the present invention clearer, the preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, wherein: Figure 1 The lung coefficient of mice after treatment with naringin; Figure 2 The expression of IL-1β mRNA in lung tissue after treatment with naringin; Figure 3 The expression of IL-6 mRNA in lung tissue after treatment with naringin; Figure 4 The expression of TNF-α mRNA in lung tissue after treatment with naringin; Figure 5 PEF (maximum respiratory rate) represents lung function after treatment with naringin. Figure 6 Lung function after naringin treatment, TV (tidal volume). Figure 7 Lung function after naringin treatment, MV (minute ventilation). Figure 8 HE staining was used to detect the pulmonary structural edema and exudation after naringin treatment. Detailed implementation manners

[0026] The present invention will be further described below in combination with the detailed implementation manners.

[0027] Example 1 Male C57BL / 6J mice around 8 weeks old and weighing 18 - 20 g were used to establish a high - altitude pulmonary edema model, provided by Jiangsu Jicui Yakang Biotechnology Co., Ltd., and the experimental animal license number was SCXK(Su)2023 - 0009. The mice were randomly divided into five groups. The first group was the Con group, the second group was the Mod group, the third group was the Nar group, the fourth group was the NaB group, and the fifth group was the Nar + NaB group. Among them, the Nar group was intraperitoneally injected with 80 mg / kg of naringin every day, the NaB group was intraperitoneally injected with 60 mg / kg of sodium butyrate every day, and the Nar + NaB group was intraperitoneally injected with 80 mg / kg of naringin + 60 mg / kg of sodium butyrate every day. The Con group and the Mod group were injected with an equal volume of normal saline every day for 7 consecutive days. Subsequently, the Mod group, the Nar group, the NaB group, and the Nar + NaB group were exposed in a small animal hypobaric oxygen chamber for 48 hours to simulate an altitude of 6000 meters to establish a mouse high - altitude pulmonary edema model. After the model was established, the mice were sacrificed by cervical dislocation, and the intact lung tissues of the mice were weighed, and the lung coefficient of the mice was calculated.

[0028] Figure 1 Shown is the lung coefficient of mice after naringin treatment. Figure 1 It can be seen from the results that naringin treatment can reduce the lung coefficient of mice in the 6000 - meter 48 - hour high - altitude pulmonary edema model, and the combination of naringin and sodium butyrate has a better prevention and treatment effect. Therefore, naringin and the composition of naringin and sodium butyrate can reduce the lung coefficient induced by the mouse high - altitude pulmonary edema model.

[0029] Example 2 Male C57BL / 6J mice weighing about 18 - 20 g at 8 weeks old were used to establish a high altitude pulmonary edema model, provided by Jiangsu Jicui Yakang Biotechnology Co., Ltd., with the experimental animal license number SCXK(Su)2023 - 0009. The mice were randomly divided into five groups. The first group was the Con group, the second group was the Mod group, the third group was the Nar group, the fourth group was the NaB group, and the fifth group was the Nar+NaB group. Among them, the Nar group was intraperitoneally injected with 80 mg / kg naringin every day, the NaB group was intraperitoneally injected with 60 mg / kg sodium butyrate every day, and the Nar+NaB group was intraperitoneally injected with 80 mg / kg naringin + 60 mg / kg sodium butyrate every day. The Con group and the Mod group were injected with an equal volume of normal saline every day for 7 consecutive days. Subsequently, the Mod group, Nar group, NaB group, and Nar+NaB group were exposed in a small animal low-pressure oxygen chamber for 48 hours to simulate an altitude of 6000 meters to establish a high altitude pulmonary edema model in mice. After the model was established, the lungs of the mice were taken for Rt-PCR to detect the expression of inflammatory factors in the lungs of the mice.

[0030] For real-time fluorescence quantitative PCR (real-time quantitative polymerase chain reaction, qRT-PCR), 20 mg of mouse lung tissue was taken and immersed in 300 μL of TRIZOL. Three sterilized steel beads were added, and a homogenizer was used for homogenization. After homogenization, 700 μL of TRIZOL was added, then 300 μL of chloroform was added, and it was shaken on a shaker at 4°C for 5 min, and then centrifuged at 12000 rpm for 30 min. 450 μL of the supernatant was taken, an equal volume of isopropanol was added, and it was alcohol-precipitated overnight at -20°C. Centrifuged at 12000 rpm at 4°C for 15 min, the supernatant was aspirated and discarded, and the precipitate was resuspended and washed with 1 mL of 70% volume concentration alcohol at 12000 rpm for 2 min. The precipitate was resuspended and washed again with 1 mL of 70% volume concentration alcohol at 12000 rpm for 2 min. After air-drying the precipitate in a fume hood, it was dissolved with DEPC water. After measuring the RNA concentration, it was reverse transcribed into cDNA according to the instructions of the RNA reverse transcription kit. Three replicate wells were set for each gene, with 1 μL of primer in each well (synthesized by BGI Genomics Co., Ltd., and the primer sequences are shown in Table 1), 1 μL of cDNA, 8 μL of distilled water, and 10 μL of mix. The qRT-PCR was detected using the Roche Light Cycler 480II system, and the reaction conditions were 95°C for 2 min; 95°C for 15 s, 60°C for 40 s, repeated for 50 cycles.

[0031] Table 1 mIL-6-RT-F 5'-CTGCAAGAGACTTCCATCCAG-3' SEQ ID NO.1 mIL-6-RT-R 5'-AGTGGTATAGACAGGTCTGTTGG-3' SEQ ID NO.2 mTNF-α-RT-F 5'-ACTGAACTTCGGGGTGATCGG-3' SEQ ID NO.3 mTNF-α-RT-R 5'-GGCTTGTCACTCGAATTTTGAGA-3' SEQ ID NO.4 m-IL-1β-F 5'-AAATACCTGTGGCCTTGGGC-3' SEQ ID NO.5 m-IL-1β-R 5'-CTTGGGATCCACACTCTCCAG-3' SEQ ID NO.6 Figures 2-4 Shows the mRNA expression of TNF-α, IL-1β and IL-6 in lung tissue after naringin treatment, by Figures 2-4 As can be seen from the results, murine high altitude pulmonary edema leads to the expression of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissue. Naringin treatment can significantly reduce the expression of TNF-α, IL-1β and IL-6 inflammatory factors in the lungs. The combination of sodium butyrate and naringin has a more obvious effect on reducing inflammatory factors. Moreover, the expression of inflammatory factors TNF-α, IL-1β and IL-6 is a key factor in the inflammation caused by high altitude pulmonary edema. Therefore, naringin can inhibit the expression of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissue, and the combination of naringin and sodium butyrate has a better effect.

[0032] Example 3 Male C57BL / 6J mice around 8 weeks old and weighing 18 - 20 g were used to establish a high altitude pulmonary edema model, provided by Jiangsu Jicui Yakang Biotechnology Co., Ltd., with the experimental animal license number SCXK(Su)2023 - 0009. The mice were randomly divided into five groups: the first group was the Con group, the second group was the Mod group, the third group was the Nar group, the fourth group was the NaB group, and the fifth group was the Nar+NaB group. Among them, the Nar group was intraperitoneally injected with 80 mg / kg of naringin every day, the NaB group was intraperitoneally injected with 60 mg / kg of sodium butyrate every day, and the Nar+NaB group was intraperitoneally injected with 80 mg / kg of naringin + 60 mg / kg of sodium butyrate every day. The Con group and the Mod group were injected with an equal volume of normal saline every day for 7 consecutive days. Subsequently, the Mod group, Nar group, NaB group, and Nar+NaB group were exposed in a small animal low-pressure oxygen chamber for 48 hours to simulate an altitude of 6000 meters to establish a murine high altitude pulmonary edema model. After the model was established, the lung function of the mice was measured.

[0033] Figures 5-7 The following shows the lung function after naringin treatment. Figures 5-7 As can be seen from the results, murine high altitude pulmonary edema causes impairment of lung function. Specifically, after the successful establishment of the acute pulmonary edema model, the maximum expiratory volume, minute ventilation volume, and tidal volume in the lung function of the mice all decreased, while naringin can improve lung function. Therefore, naringin can alleviate the reduction of lung function, and the combination of naringin and sodium butyrate has a better effect.

[0034] Example 4 Male C57BL / 6J mice weighing about 18 - 20 g at about 8 weeks old were used to establish a high altitude pulmonary edema model, provided by Jiangsu Jicui Yakang Biotechnology Co., Ltd., and the experimental animal license number is SCXK(Su)2023 - 0009. The mice were randomly divided into five groups. The first group was the Con group, the second group was the Mod group, the third group was the Nar group, the fourth group was the NaB group, and the fifth group was the Nar + NaB group. Among them, the Nar group was intraperitoneally injected with 80 mg / kg of naringin every day, the NaB group was intraperitoneally injected with 60 mg / kg of sodium butyrate every day, and the Nar + NaB group was intraperitoneally injected with 80 mg / kg of naringin + 60 mg / kg of sodium butyrate every day. The Con group and the Mod group were injected with an equal volume of normal saline every day for 7 consecutive days. Subsequently, the Mod group, Nar group, NaB group, and Nar + NaB group were exposed in a small animal low - pressure oxygen chamber for 48 hours to simulate an altitude of 6000 meters to establish a high altitude pulmonary edema model in mice. After the model was established, the lung tissues of the mice were taken for HE staining to detect the edema and exudation of the lung tissue structure in the mice.

[0035] Figure 8 Shown is the detection of the edema and exudation of the lung structure after naringin treatment by HE staining, by Figure 8 As can be seen from the results, high altitude pulmonary edema in mice led to the occurrence of edema and exudation in the lung tissue structure. Specifically, on the one hand, the high altitude pulmonary edema model in mice directly caused an acute inflammatory response in the lungs. On the other hand, during the occurrence of pulmonary edema, pulmonary vasoconstriction caused a sharp increase in pulmonary artery pressure, damaged the pulmonary capillary endothelial barrier, and led to the occurrence of an acute inflammatory response. And naringin could inhibit the occurrence of edema and exudation in the lung tissue structure caused by the high altitude pulmonary edema model in mice. Therefore, naringin could inhibit high altitude pulmonary edema induced by the high altitude pulmonary edema model in mice, and the combination of naringin and sodium butyrate had a better effect.

[0036] It can be seen from Examples 1 - 4 that drugs for preventing and treating high altitude pulmonary edema prepared from naringin and the combination of naringin and sodium butyrate can reduce high altitude pulmonary edema in mice, specifically including: reducing the lung coefficient; inhibiting the expression of inflammatory factors TNF - α, IL - 1β and IL - 6 in the lung tissue; improving lung function; blocking the occurrence of edema and exudation in the lung tissue structure. Therefore, naringin can be used as a potential drug for preventing and treating high altitude pulmonary edema.

[0037] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the purpose and scope of the present technical solution, and they should all be covered within the scope of the claims of the present invention.

Claims

1. Application of naringin in the preparation of drugs for the prevention and treatment of high-altitude pulmonary edema.

2. The application according to claim 1, characterized in that, The plateau has an altitude of over 6,000 meters.

3. The application according to claim 1, characterized in that, Naringin helps prevent high-altitude pulmonary edema by inhibiting the expression of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissue.

4. The application according to claim 1, characterized in that, Naringin can prevent and treat high-altitude pulmonary edema by improving lung function.

5. A combination of naringin and sodium butyrate.

6. The composition according to claim 5, characterized in that, The mass ratio of naringin to sodium butyrate is 4:

3.

7. The use of the composition according to claim 5 in the preparation of a drug for preventing and treating high-altitude pulmonary edema.

8. A drug for the prevention and treatment of high-altitude pulmonary edema, characterized in that, Its active ingredient is naringin.

9. A drug for the prevention and treatment of high-altitude pulmonary edema, characterized in that, It comprises the composition of claim 5.

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