Application of ginsenoside Re in preparation of medicine for treating acute lung injury related to severe acute pancreatitis

Abstract

The invention relates to the field of novel application of ginsenoside Re, in particular to application of ginsenoside Re in preparation of medicines for treating acute lung injury related to severe acute pancreatitis. By constructing an in-vitro endothelial cell injury model, it is verified that ginsenoside Re can reduce LPS-induced cell death, can reduce cell leakage and can reduce endothelial connection destroyed by LPS, and the protective effect of ginsenoside Re on endothelial cells in SAP-ALI is verified. An SAP animal model experiment verifies that ginsenoside Re can protect pancreas of an SAP-ALI mouse from being damaged and can also protect a pulmonary vessel barrier of the SAP-ALI mouse at the same time. Multiple verifications show that the application of ginsenoside Re has important clinical significance on treatment of patients with serious complications SAP-ALI, and a new strategy and hope are provided for treatment of SAP-ALI in the future.

Description

Technical Field

[0001] The present invention relates to a new application field of ginsenoside Re, and in particular to the application of ginsenoside Re in the preparation of a drug for treating acute lung injury associated with severe acute pancreatitis. Background Art

[0002] Acute lung injury (ALI) is a serious complication of severe acute pancreatitis (SAP). Its pathophysiological characteristics are: alveolar-capillary damage. ALI is usually caused by systemic inflammatory response syndrome (SIRS), which leads to damaged barrier function of alveoli and capillaries. Damage to alveolar epithelial cells and capillary endothelial cells destroys the normal gas exchange interface, resulting in reduced exchange efficiency of oxygen and carbon dioxide; pulmonary edema. The damaged alveolar-capillary barrier allows fluid, protein and leukocytes to infiltrate from blood vessels into alveoli and interstitium, forming pulmonary edema. Pulmonary edema further aggravates gas exchange disorders and makes it impossible for alveoli to be oxygenated effectively; hypoxemia. Due to damage to alveoli and capillaries and pulmonary edema, the oxygen content in the blood decreases, leading to hypoxemia. Hypoxemia prevents other organs and tissues in the body from getting enough oxygen supply, which may lead to organ dysfunction. Clinical manifestations: Due to gas exchange disorders, patients experience shortness of breath and dyspnea; blood gas analysis shows a decrease in arterial oxygen partial pressure (PaO2), usually below 60 mmHg, even with oxygen inhalation; diffuse infiltration of both lungs, which is due to pulmonary edema and inflammatory response; in the severe stage of ALI, acute respiratory distress syndrome (ARDS) may develop, which is a more serious state of respiratory failure.

[0003] The treatment of ALI includes providing adequate oxygen support, using mechanical ventilation when necessary, and treatment of the primary cause. The prognosis of ALI depends on the severity of the injury, the patient's overall health, and whether appropriate treatment is received in a timely manner. Severe ALI may lead to ARDS and have a poor prognosis. Acute lung injury is one of the important causes of death in patients with severe acute pancreatitis. Therefore, timely identification and treatment of ALI is crucial to improving the prognosis of SAP patients.

[0004] The current treatment of SAP-ALI is limited to organ function support therapy, such as oxygen inhalation, mechanical ventilation, etc., and its efficacy is limited. It is urgent to find a drug to prevent SAP-ALI and block its progression. Summary of the invention

[0005] The purpose of the present invention is to address the problem that the existing treatment methods for SAP-ALI are limited to organ function support treatment and have limited efficacy, and to propose the use of ginsenoside Re in the preparation of a drug for treating severe acute pancreatitis-related acute lung injury. Studies have found that ginsenoside Re has great clinical transformation value in preventing SAP-ALI and blocking its progression.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] The first purpose of the present invention is to protect the use of ginsenoside Re in the preparation of drugs for preventing or treating acute lung injury associated with severe acute pancreatitis.

[0008] The second purpose of the present invention is to protect the use of ginsenoside Re in the preparation of drugs for improving pulmonary oxidative stress damage.

[0009] The third purpose of the present invention is to protect the use of ginsenoside Re in the preparation of drugs for protecting pulmonary vascular barrier function.

[0010] The inventors verified that ginsenoside Re can reduce LPS-induced cell death, cell leakage and LPS damage to endothelial junctions by constructing an in vitro SAP model, and verified the protective effect of ginsenoside Re on SAP-ALI. Through SAP animal model experiments, it was verified that ginsenoside Re protects pancreatic damage in SAP-ALI mice and can also protect the pulmonary vascular barrier in SAP-ALI mice. Multiple verifications show that the application of ginsenoside Re has important clinical significance for the treatment of patients with severe complications of SAP-ALI, and provides new strategies and hopes for the future treatment of SAP-ALI.

[0011] Another object of the present invention is to protect the application of the pharmaceutical composition comprising ginsenoside Re.

[0012] A use of a pharmaceutical composition in the preparation of any one or more functional drugs of the following ac, wherein the pharmaceutical composition comprises a therapeutically effective amount of ginsenoside Re and a pharmaceutically acceptable excipient;

[0013] a. Prevent or treat severe acute pancreatitis-related acute lung injury;

[0014] b. Drugs that improve lung oxidative stress damage;

[0015] c. Protect pulmonary vascular barrier function.

[0016] Furthermore, the pharmaceutically acceptable excipients include at least one of diluents, excipients, fillers, binders, wetting agents, absorption enhancers, surfactants, lubricants, stabilizers, flavoring agents, sweeteners, and pigments.

[0017] Furthermore, the pharmaceutical composition is a liquid preparation, a solid preparation or a spray preparation.

[0018] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0019] The present invention relates to a new application field of ginsenoside Re, and specifically to the application of ginsenoside Re in the preparation of a drug for treating acute lung injury associated with severe acute pancreatitis. By constructing an in vitro SAP model, it was verified that ginsenoside Re can reduce LPS-induced cell death, reduce cell leakage, and reduce LPS damage to endothelial junctions, and the protective effect of ginsenoside Re on SAP-ALI was verified. Through SAP animal model experiments, it was verified that ginsenoside Re protected pancreatic damage in SAP-ALI mice, and at the same time, it could protect the pulmonary vascular barrier in SAP-ALI mice. It has been verified from multiple sources that the application of ginsenoside Re has important clinical significance for the treatment of patients with severe complications of SAP-ALI, and provides new strategies and hopes for the future treatment of SAP-ALI. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a data graph showing that ginsenoside Re reduces LPS-induced cell death. The left graph shows the PI fluorescence value at consecutive time points / baseline PI fluorescence value; the right graph shows the statistical data at 0 hour, 3 hour, 6 hour, 9 hour, and 12 hour.

[0021] Figure 2 Data showing that ginsenoside Re reduces LPS-induced cell leakage.

[0022] Figure 3 This is the data graph showing that ginsenoside Re reduces LPS damage to endothelial-endothelial junctions.

[0023] Figure 4 This is the data chart showing that ginsenoside Re improves the condition of SAP-ALI mice. Figure 4 A in the figure indicates the construction of SAP-ALI mouse model and the administration arrangement of ginsenoside Re; Figure 4 B in the figure is the statistical graph of pancreatic pathological scores of SAP mice; Figure 4 C in the figure is the trypsin activity data of pancreatic tissue; Figure 4 D in it is the MPO activity of pancreatic tissue; Figure 4 E in the figure is the data graph of MPO activity in lung tissue.

[0024] Figure 5 This is the data of ginsenoside Re protecting the pulmonary vascular barrier of SAP-ALI mice; Figure 5 A in the figure is the data diagram of Evans blue leakage degree of lung tissue; Figure 5B in the figure is the data graph of wet-to-dry weight ratio of lung tissue. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0026] Example 1

[0027] Acute lung injury (ALI) is a serious complication of severe acute pancreatitis (SAP). The treatment of ALI includes providing adequate oxygen support, using mechanical ventilation when necessary, and treating the primary cause. The prognosis of ALI depends on the severity of the injury, the patient's overall health, and whether appropriate treatment is received in a timely manner. Severe ALI may lead to ARDS and have a poor prognosis. Acute lung injury is one of the important causes of death in patients with severe acute pancreatitis. Therefore, timely identification and treatment of ALI is crucial to improving the prognosis of SAP patients.

[0028] The current treatment of SAP-ALI is limited to organ function support therapy, such as oxygen inhalation, mechanical ventilation, etc., and its efficacy is limited. It is urgent to find a drug to prevent SAP-ALI and block its progression.

[0029] Ginsenoside Re is a tetracyclic triterpenoid derivative, colorless needle-shaped crystals. The prior art discloses that ginsenoside Re can inhibit the central nervous system and promote DNA and RNA synthesis. It increases the effect of plasma corticosterone and dilates blood vessels. It can reduce the contraction of the guinea pig's isolated uterus caused by acetylcholine. It has the effect of slowing down the heart rate and biphasic blood pressure (first rising and then falling) in rats. It has a moderate inhibitory effect on the behavior and electroencephalogram of cats. It has an anti-fatigue effect. However, the prior art has not found any research on the effect of ginsenoside Re in SAP-ALI disease.

[0030] This example proposes the use of ginsenoside Re in the preparation of a drug for treating severe acute pancreatitis-related acute lung injury. Studies have found that ginsenoside Re has great clinical translational value in preventing SAP-ALI and blocking its progression.

[0031] The details are as follows:

[0032] Some sources of raw materials in the following examples:

[0033] Ginsenoside Re was purchased from Pusi Biotechnology, product number CN000010, 100 mg.

[0034] LPS was purchased from Thermo Fisher Scientific, catalog number L2880, 10 mg.

[0035] Caerulein was purchased from TOCRIS, product number 17650-98-5, 1 mg.

[0036] Propidium iodide (PI) was purchased from Thermo Fisher Scientific, catalog number 2549278, 10 ml.

[0037] FITC-dextran was purchased from Thermo Fisher Scientific, catalog number 46944, 100 mg.

[0038] Constructing an in vitro SAP model to verify the protective effect of ginsenoside Re on SAP-ALI

[0039] (1) SAP-ALI in vitro model:

[0040] Rat pulmonary microvascular endothelial cells (PMVEC) were stimulated with lipopolysaccharide (LPS) as an in vitro endothelial cell injury model of SAP-ALI. Ginsenoside Re was co-incubated with LPS as the drug intervention group.

[0041] The propidium iodide (PI) staining status was detected using an enzyme-labeled instrument to observe the cell death ratio in each group.

[0042] After the cells were plated in 96-well plates for 24 hours, they were treated accordingly. Drug incubation: In the drug group, different concentrations of ginsenoside Re (50uM, 100uM, 200uM) were incubated in advance for 1 hour. In the LPS model group and the control group, the corresponding solvents were added and incubated for 1 hour. LPS stimulation: The drug group and the LPS model group were stimulated by adding 10ug / ml LPS, and the control group was incubated with the corresponding solvents. The staining status of propidium iodide (PI) was detected using an ELISA instrument to observe the cell death ratio in each group. Figure 1 The test results showed that ginsenoside Re could reduce LPS-induced cell death.

[0043] The concentration of FITC-dextran in the lower chamber of the transwell was detected by an enzyme-labeled instrument to observe the degree of cell leakage in each group.

[0044] After the cells were plated in the upper chamber of the transwell for 72 hours, they were treated accordingly. Drug incubation: In the drug group, different concentration gradients of ginsenoside Re (50uM, 100uM, 200uM) were incubated in advance for 1 hour. In the LPS model group and the control group, the corresponding solvents were added and incubated for 1 hour. LPS stimulation: The drug group and the LPS model group were stimulated by adding 10ug / ml LPS, and the control group was incubated with the corresponding solvents. After 12 hours, the liquid in the upper chamber was replaced with a solution containing FITC-dextran, and the lower chamber was replaced with a PBS solution without FITC-dextran. The cells were incubated in the dark for half an hour, and the FITC-dextran concentration in the lower chamber of the transwell was detected using an enzyme reader to observe the degree of cell leakage in each group. Figure 2 The test results showed that ginsenoside Re could reduce LPS-induced cell leakage.

[0045] RT-qPCR technology was used to detect endothelial cell junction molecules and observe the transcriptional expression levels of endothelial cell junction molecules in each group.

[0046] After the cells were plated in 6-well plates for 24 hours, they were treated accordingly. Drug incubation: In the drug group, different concentrations of ginsenoside Re (50uM, 100uM, 200uM) were incubated in advance for 1 hour. In the LPS model group and the control group, the corresponding solvents were added and incubated for 1 hour. LPS stimulation: The drug group and the LPS model group were stimulated by adding 10ug / ml LPS, and the control group was incubated with the corresponding solvents. After 12 hours, the cells were collected, and the cell RNA was extracted according to the instructions, followed by reverse transcription and qPCR. Figure 3 The test results showed that ginsenoside Re reduced LPS-induced damage to endothelial inter-endothelial junctions.

[0047] (2) SAP animal model:

[0048] SAP-ALI mouse animal model: Mice were intraperitoneally injected with cerulein (CER; 100ug / kg) for 10 consecutive times, with an interval of 1 hour between each injection. Immediately after the last injection of CER, an injection of lipopolysaccharide (LPS; 10mg / kg) was given.

[0049] Ginsenoside Re administration method: The time of the first CER injection in the SAP-ALI mouse animal model was recorded as the first hour, and ginsenoside Re was injected intraperitoneally at the 3rd, 6th, 9th, 15th, 21st, and 27th hours after the model was established. The mice in the control group and the model group were injected with normal saline according to the same arrangement. 33 hours after the first CER injection, the mice were anesthetized and killed, and pancreatic tissue, lung tissue, and mouse serum were collected for subsequent testing, including pancreatic pathology, pancreatic trypsin activity, pancreatic MPO activity, and lung MPO activity. Figure 4 The results showed that ginsenoside Re protected SAP-ALI mice from pancreatic damage, as shown by improved pancreatic pathology, decreased pancreatic trypsin and pancreatic MPO, and alleviated SAP-ALI damage, as shown by decreased lung MPO. In addition, ginsenoside Re also protected the pulmonary vascular barrier in SAP-ALI mice, as shown by decreased Evans blue leakage in lung tissue and decreased wet-to-dry weight ratio in lung tissue ( Figure 5 ).

[0050] The present invention relates to a new application field of ginsenoside Re, and specifically to the application of ginsenoside Re in the preparation of a drug for treating acute lung injury associated with severe acute pancreatitis. By constructing an in vitro SAP model, it was verified that ginsenoside Re can reduce LPS-induced cell death, reduce cell leakage, and reduce LPS damage to endothelial junctions, and the protective effect of ginsenoside Re on SAP-ALI was verified. Through SAP animal model experiments, it was verified that ginsenoside Re protected pancreatic damage in SAP-ALI mice, and at the same time, it could protect the pulmonary vascular barrier in SAP-ALI mice. It has been verified from multiple sources that the application of ginsenoside Re has important clinical significance for the treatment of patients with severe complications of SAP-ALI, and provides new strategies and hopes for the future treatment of SAP-ALI.

[0051] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of ginsenoside Re in the preparation of drugs for preventing or treating acute lung injury associated with severe acute pancreatitis.

2. Application of ginsenoside Re in the preparation of drugs for improving pulmonary oxidative stress damage.

3. Application of ginsenoside Re in the preparation of drugs for protecting pulmonary vascular barrier function.

4. Use of a pharmaceutical composition in the preparation of any one or more functional drugs of the following ac, characterized in that: The pharmaceutical composition comprises a therapeutically effective amount of ginsenoside Re and pharmaceutically acceptable excipients; a. Prevent or treat severe acute pancreatitis-related acute lung injury; b. Drugs that improve lung oxidative stress damage; c. Protect pulmonary vascular barrier function.

5. The use according to claim 4, characterized in that: Pharmaceutically acceptable excipients include at least one of diluents, excipients, fillers, binders, wetting agents, absorption enhancers, surfactants, lubricants, stabilizers, flavoring agents, sweeteners, and pigments.

6. The use according to claim 4 or 5, characterized in that: The pharmaceutical composition is a liquid preparation, a solid preparation or a spray preparation.

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