CN106727504B

CN106727504B - Application of simvastatin in preparation of medicine for preventing pulmonary oxygen poisoning caused by high partial pressure oxygen

Info

Publication number: CN106727504B
Application number: CN201611056076.1A
Authority: CN
Inventors: 包晓辰; 方以群; 攸璞; 王芳芳; 马骏; 谷爱梅
Original assignee: Navy Medicine Research Institute of PLA
Current assignee: Navy Medicine Research Institute of PLA
Priority date: 2016-11-24
Filing date: 2016-11-24
Publication date: 2020-04-24
Anticipated expiration: 2036-11-24
Also published as: CN106727504A

Abstract

The invention belongs to the field of medicine preparation, and particularly relates to application of simvastatin in preparation of a medicine for preventing pulmonary oxygen poisoning caused by high partial pressure oxygen, wherein simvastatin is dissolved in physiological saline to be prepared into an injection of 0.5-2 mg/mL. The dosage of the simvastatin for intraperitoneal injection is 5-20mg/Kg/d, and the administration time is 3 days before high-partial-pressure oxygen exposure, and the simvastatin is taken once a day. Experiments prove that simvastatin can remarkably reduce the increase of lung permeability caused by high partial pressure oxygen, reduce the content of protein in pulmonary edema and bronchoalveolar lavage fluid, relieve lung inflammation and reduce the proportion of apoptosis of lung cells, thereby relieving lung injury caused by lung type oxygen poisoning. Moreover, the invention provides experimental data for more reasonably applying simvastatin in prevention of pulmonary oxygen poisoning caused by high partial pressure oxygen clinically.

Description

Application of simvastatin in preparation of medicine for preventing pulmonary oxygen poisoning caused by high partial pressure oxygen

Technical Field

The invention belongs to the field of medicine preparation, and particularly relates to application of simvastatin in preparation of a medicine for treating pulmonary oxygen poisoning caused by high partial pressure oxygen.

Background

When an oxygen breathing device is adopted in clinical application of a breathing machine for treating respiratory failure patients, hyperbaric oxygen for treating decompression sickness and diving operation, the occurrence of pulmonary oxygen poisoning can be caused because the ambient air pressure is too high (or the diving depth is too large), the oxygen concentration is too high and the time for inhaling 60-200kPa hyperbaric oxygen is too long, which exceeds the adaptive capacity of the organism. The lung is the primary target organ for oxygen poisoning, since the respiratory system is exposed to higher partial pressures of oxygen than other organs, while being relatively sensitive to oxygen. The main clinical manifestations of pulmonary type oxygen poisoning are cough, pain behind the sternum, shortness of breath, and severe cases with pulmonary edema, hemorrhage, and even respiratory failure. Thereby influencing the diving operation and the application of clinical high partial pressure oxygen.

The pathogenesis of oxygen poisoning has not yet been fully elucidated. Previous studies have shown that oxygen metabolites in the body, oxygen radicals, may be the initiating factor in the development of pulmonary-type oxygen toxicity. When the body inhales excessive oxygen, the generated oxygen radicals exceed the metabolic capability of antioxidant enzymes in the body, and the accumulated free radicals have harmful effects on various organs. Although a great deal of research is conducted at home and abroad, the prevention and treatment of lung-type oxygen poisoning is still very limited at present: the research on the mechanism of oxygen free radicals damaging alveolar epithelial cells is not completely clear, and a specific medicine for preventing and treating pulmonary oxygen poisoning is lacking clinically.

Simvastatin is an HMG-COA reductase inhibitor, can inhibit the synthesis of endogenous cholesterol, and plays a role in reducing blood fat. Is suitable for patients with hyperlipidemia, coronary heart disease complicated with hypercholesterolemia and arteriosclerosis. However, no study has confirmed that it can be used for preventing pulmonary oxygen poisoning.

Disclosure of Invention

The invention aims to provide application of simvastatin in a medicament for preventing pulmonary type oxygen poisoning caused by high partial pressure oxygen, wherein the simvastatin can obviously reduce lung permeability, lung tissue injury, inflammatory reaction and lung epithelial cell apoptosis of the pulmonary type oxygen poisoning caused by high partial pressure oxygen.

The technical scheme provided by the invention is that simvastatin is used for preparing a medicine for preventing pulmonary oxygen poisoning caused by high partial pressure oxygen.

For the application, simvastatin needs to be dissolved in physiological saline to be prepared into an injection of 0.5-2 mg/mL.

The dosage of the simvastatin for intraperitoneal injection is 5-20 mg/kg/d. Preferably, the dosage of the simvastatin for intraperitoneal injection is 20 mg/kg/d.

The simvastatin was used 3 days before high partial pressure oxygen exposure, once a day. The last injection was 1h before high partial pressure oxygen exposure.

The invention has the beneficial effects that:

1. the invention provides application of simvastatin in preparation of a drug for treating pulmonary oxygen poisoning caused by high partial pressure oxygen, and experiments show that simvastatin can remarkably reduce increase of lung permeability caused by high partial pressure oxygen, reduce pulmonary edema and protein content in bronchoalveolar lavage fluid, relieve pulmonary inflammation and reduce the proportion of apoptosis of lung cells, thereby relieving lung injury caused by pulmonary oxygen poisoning.

2. The invention provides experimental data for more reasonably applying simvastatin in prevention of pulmonary oxygen poisoning caused by high partial pressure oxygen clinically.

Drawings

FIG. 1 is a graph comparing the effect of different doses of simvastatin used prophylactically in example 1 on the lung wet-to-dry weight ratio in a mouse pulmonary type oxygen toxicity model caused by high partial pressure of oxygen.

FIG. 2 is a graph comparing the effect of preventive use of simvastatin on lung tissue damage in the mouse lung-type oxygen intoxication model caused by high partial pressure of oxygen in example 1. Wherein, the graph A is a pathological graph of lung tissues of mice in an air exposure and normal saline group; figure B is a pathological graph of lung tissue in mice with high partial pressure oxygen exposure plus normal saline; and the graph C is a pathological graph of simvastatin 20mg/Kg for preventing lung tissues of mice in a group, and the magnification is 200 times.

FIG. 3 is a graph comparing the change in lung permeability in the mouse lung type oxygen toxicity model caused by high partial pressure of oxygen in the preventive use of simvastatin in example 2, wherein A is the wet-to-dry weight ratio of lung tissue; panel B shows the protein content of alveolar lavage fluid.

FIG. 4 is a graph comparing the changes of pro-inflammatory factor of lung tissue and apoptotic protein in the mouse lung type oxygen toxicity model caused by high partial pressure of oxygen in example 2 in the preventive use of simvastatin, wherein panel A is pro-inflammatory factor of lung tissue; panel B is lung tissue apoptotic protein.

FIG. 5 shows that in example 3, the L-NAME inhibitor can completely eliminate the protective effect of simvastatin on mouse pulmonary oxygen toxicity caused by high partial pressure of oxygen, wherein FIG. A shows that simvastatin can increase the expression level of eNOS enzyme in mouse lung tissues after being used; FIG. B is a graph of the effect of L-NAME on the wet/dry ratio of lung tissue and protein content in alveolar lavage fluid after application; panel C is the effect of L-NAME on lung tissue apoptotic protein expression after application; panel D shows the effect of L-NAME on proinflammatory factors in lung tissue after application.

Detailed Description

The invention will now be further illustrated by reference to the following examples:

simvastatin used in the experiments of the present invention was purchased from msandong.

Example 1

Injection medicine for experiment: simvastatin is prepared into a solution of 0.5-2mg/mL by using normal saline, and is prepared into an injection.

Experimental animals and groups: male healthy C57BL/6 mice, 50, weighing 20-25g, were purchased from Shanghai Spiker laboratory animals, Inc.

Mice were randomly divided into an air-exposed + saline group, a hyperbaric oxygen-exposed + simvastatin 5mg/Kg group, a hyperbaric oxygen-exposed + simvastatin 10mg/Kg group, and a hyperbaric oxygen-exposed + simvastatin 20mg/Kg group, with 10 mice per group.

The experimental method comprises the following steps: 3 days before high partial pressure oxygen exposure, simvastatin (5, 10 and 20mg/kg) is respectively injected into the abdominal cavity of a simvastatin group, and an equal volume of normal saline is injected into the abdominal cavity of a control group. After injection on day 3, the experimental animals were placed in an animal oxygen chamber for 1 hour, pressurized to 0.23MPa, and after 100% oxygen exposure for 6 hours, the animals were evacuated from the chamber within 2 minutes. And taking lung tissues of the mice for detection.

As shown in FIG. 1, the wet-to-dry weight ratio of the lung tissue of the mice in the air-exposed control group was 4.47. + -. 0.11 (mean. + -. standard deviation), and the wet-to-dry weight ratio of the high partial pressure oxygen-exposed + physiological saline group was 4.81. + -. 0.23, which was significantly higher than that in the air-exposed control group, indicating an increase in lung permeability. The wet-dry weight ratio of the high partial pressure oxygen exposure plus simvastatin 5mg/Kg group is 4.62 plus or minus 0.15, the 10mg/Kg group is 4.59 plus or minus 0.14, and the 20mg/Kg group is 4.49 plus or minus 0.09. Wherein the wet-dry weight ratio of 20mg/Kg group of lung is obviously reduced compared with the group of high partial pressure oxygen exposure and normal saline, and the p value is less than 0.05, which shows that the lung permeability can be obviously reduced by the pretreatment of 20mg/Kg of simvastatin.

The pathological examination of lung tissues of mice in each group shows that the pulmonary alveolar structure of the mice in the high partial pressure oxygen exposure and normal saline group is destroyed, the edema of the alveolar wall is obvious, and eosinophilic red liquid and erythrocyte exudation can be seen in the alveolar cavity. In the simvastatin intervention group, the conditions of mouse pulmonary alveolar structure damage, alveolar wall edema and hemorrhage in an alveolar cavity are obviously reduced. In particular, as shown in fig. 2, wherein, a is a pathological diagram of lung tissues of mice in an air exposure + physiological saline group; figure B is a pathological graph of lung tissue in mice with high partial pressure oxygen exposure plus normal saline; and the graph C is a pathological graph of simvastatin 20mg/Kg for preventing lung tissues of mice in a group, and the magnification is 200 times.

And (4) conclusion: by comparing the intervention effect of simvastatin with different doses on pulmonary type oxygen poisoning caused by high partial pressure oxygen, 20mg/Kg intervention of simvastatin is shown to be the best preferential dose, so in the subsequent experiments, 20mg/Kg is adopted as the intervention dose of the experiment.

Example 2

Injection medicine for experiment: simvastatin is prepared into a solution of 2mg/mL by using normal saline, and is prepared into an injection.

Experimental animals and groups: male healthy C57BL/6 mice, 40, weighing 20g, were purchased from Shanghai Spiker laboratory animals, Inc.

Mice were randomly divided into an air-exposed + saline group, an air-exposed + simvastatin group, a hyperbaric oxygen-exposed + saline group, and a hyperbaric oxygen-exposed + simvastatin group, with 10 mice per group.

The experimental method comprises the following steps: 3 days before high partial pressure oxygen exposure, simvastatin (20mg/kg) was injected intraperitoneally in the simvastatin group, and an equal volume of physiological saline was injected intraperitoneally in the control group. After injection on day 3, the experimental animals were placed in an animal oxygen chamber for 1 hour, pressurized to 0.23MPa, and after 100% oxygen exposure for 6 hours, the animals were evacuated from the chamber within 2 minutes. And taking lung tissues of the mice for detection.

The experimental result shows that simvastatin can obviously reduce the increase of lung permeability caused by high partial pressure of oxygen. Compared with the air exposed group, the wet-dry specific gravity of the lung tissue and the protein content in the alveolar lavage fluid of the mice in the hyperbaric oxygen exposed + normal saline group are obviously increased, and the simvastatin intervention can obviously reduce the wet-dry specific gravity of the lung tissue and the protein content in the alveolar lavage fluid (figure 3).

As shown in FIG. 4, compared with the air-exposed group, the amounts of proinflammatory factors, TNF- α, IL-6 and IL-1 β in lung tissues of mice in hyperbaric oxygen exposure + normal saline group were significantly increased, the expression level of TUNEL positive cell number was significantly increased, and the amount of proinflammatory factors and apoptosis proteins in lung tissues of mice was significantly reduced by preventive use of simvastatin.

And (4) conclusion: the simvastatin has an obvious protective effect on the pulmonary type oxygen poisoning caused by high partial pressure oxygen in preventive use, and can reduce the increase of the permeability of lung tissues caused by the high partial pressure oxygen and reduce the expression of proinflammatory factors and apoptosis proteins.

Example 3

Injection medicine for experiment: simvastatin is prepared into a solution of 2mg/mL by using normal saline, and is prepared into an injection. Nitric oxide synthase inhibitor (L-NAME) was formulated into 2mg/mL injection using physiological saline.

Mice were randomly divided into an air exposure + saline group, a hyperbaric oxygen exposure + simvastatin + L-NAME group, 10 mice per group.

The experimental method comprises the following steps: 3 days before high partial pressure oxygen exposure, simvastatin (20mg/kg) and simvastatin + L-NAME groups were injected intraperitoneally, and an equal volume of normal saline was injected intraperitoneally in a control group. On day 3, 1h before hyperbaric oxygen exposure, the simvastatin + L-NAME group was injected intraperitoneally with L-NAME (20 mg/Kg). After injection, the experimental animal is placed in an animal oxygen chamber for 1h, the pressure is increased to 0.23MPa, 100 percent oxygen is exposed for 6h, and the animal is decompressed and taken out of the chamber within 2 minutes. And taking lung tissues of the mice for detection.

The experimental result shows that the L-NAME can completely eliminate the protective effect of simvastatin on pulmonary oxygen poisoning caused by high partial pressure oxygen.

FIG. 5 shows that the wet-to-dry weight ratio and the bronchoalveolar lavage fluid protein content of the lung tissues of the mice in the L-NAME group are significantly increased compared with the hyperbaric oxygen exposure + simvastatin group, and there is no significant difference with the hyperbaric oxygen exposure + physiological saline group.

FIG. 5 shows that the wet/dry ratio of lung tissue, the protein content in alveolar lavage fluid, the contents of pro-inflammatory factors TNF- α and IL-6 and the content of apoptosis protein clear-caspase-3 in the L-NAME group mice were significantly increased compared to the hyperbaric oxygen exposure + simvastatin group, and there was no significant difference from the hyperbaric oxygen exposure + physiological saline group.

And (4) experimental conclusion: eNOS plays a critical regulatory role in the protection of simvastatin against hyperbaric oxygen-induced pulmonary oxygen poisoning.

In conclusion, the experiments prove that simvastatin can reduce lung permeability, relieve lung inflammation and reduce apoptosis rate of lung tissue cells by up-regulating eNOS expression and activity, thereby protecting lung-type oxygen poisoning.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention, and the present invention should not be limited by the disclosure of the preferred embodiments. Therefore, it is intended that all equivalents and modifications which do not depart from the spirit of the invention disclosed herein are deemed to be within the scope of the invention.

Claims

1. Application of simvastatin in preparation of drugs for preventing pulmonary type oxygen poisoning caused by high partial pressure oxygen.

2. Use according to claim 1, characterized in that: the medicine for preventing the lung type oxygen poisoning caused by high partial pressure oxygen is an injection.

3. Use according to claim 2, characterized in that: the effective concentration of simvastatin in the injection is 0.5-2 mg/mL.

4. Use according to claim 1, characterized in that: the dosage of the simvastatin for intraperitoneal injection is 5-20 mg/kg/d.

5. Use according to claim 1, characterized in that: the simvastatin was used 3 days before high partial pressure oxygen exposure, once a day.