Application of terbutaline sulfate in preventing and treating intestinal ischemia-reperfusion injury

Abstract

The present invention belongs to the field of biotechnology, and specifically relates to the application of terbutaline sulfate in preventing and treating intestinal ischemia-reperfusion injury. The terbutaline sulfate is an oral solution of terbutaline sulfate; the terbutaline sulfate prevents and treats intestinal ischemia-reperfusion injury by reducing pathological damage to the small intestine and improving the self-renewal and repair ability of intestinal stem cells. The present invention uses terbutaline sulfate to prevent and treat intestinal ischemia-reperfusion injury, and the effect is significant. Oral administration before surgery can effectively improve intestinal ischemia-reperfusion injury indicators; the side effects are small, the excitatory effect on the heart is very small, and there is no central effect; it is easy to use, is a commonly used clinical drug, can be atomized and inhaled during the perioperative period, and has a fast onset of action; the efficacy is diverse, it is a common asthma treatment drug, can improve patient ventilation, and improve prognosis.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to application of terbutaline sulfate in preventing and treating intestinal ischemia-reperfusion injury. Background Art

[0002] Intestinal ischemia-reperfusion injury is a common emergency and critical condition. It not only damages the intestinal barrier, but also causes extraintestinal multi-organ damage, with very high complications and mortality. Therefore, prevention and treatment of intestinal ischemia-reperfusion injury is of great significance to improve the prognosis of patients. Among them, patient factors, anesthesia factors and surgery-related factors can all cause intestinal ischemia-reperfusion injury to varying degrees.

[0003] In recent years, people have conducted extensive research on drugs and strategies for preventing and treating intestinal ischemia-reperfusion injury. Currently, several strategies and strategies have been shown to reduce intestinal ischemia-reperfusion injury in small animals. Possible strategies for treating intestinal ischemia-reperfusion injury include the following: ischemic preconditioning, antioxidants, nitric oxide supplementation, anti-complement therapy, anti-leukocyte therapy, perfluorocarbons, enteral feeding, glutamine supplementation, and glycine supplementation, etc. However, the underlying mechanism of intestinal ischemia-reperfusion injury has not been fully elucidated, and effective prevention and treatment strategies and drugs need to be further explored; there is also a lack of effective biomarkers for intestinal ischemia-reperfusion injury in clinical practice.

[0004] Among them, ischemic preconditioning has been found to be the most promising strategy to combat reperfusion injury and appears to increase the intestinal tolerance to reperfusion injury. Ischemic preconditioning refers to the brief exposure of tissues to ischemic conditions, which can protect them from the harmful effects of long-term intestinal ischemia-reperfusion injury. However, the ideal ischemic preconditioning regimen, including ischemic time and number of cycles, has not yet been established. Some studies have reported that more than one ischemic and reperfusion cycle is required to achieve this beneficial effect. At the same time, although ischemic preconditioning has been shown to be beneficial to the human heart and liver, prospective controlled studies involving intestinal ischemic preconditioning in humans are still lacking. Therefore, there are still many limitations and uncertainties in the clinical application of strategies such as ischemic preconditioning.

[0005] It can be seen that there is still a lack of effective drugs and strategies for the prevention and treatment of intestinal ischemia-reperfusion injury. Summary of the invention

[0006] The purpose of the present invention is to provide an application of terbutaline sulfate in preventing and treating intestinal ischemia-reperfusion injury, effectively improving intestinal ischemia-reperfusion injury indicators and improving prognosis.

[0007] The objective of the present invention is achieved through the following technical solutions:

[0008] The invention provides an application of terbutaline sulfate in preventing and treating intestinal ischemia-reperfusion injury.

[0009] Furthermore, the terbutaline sulfate is terbutaline sulfate oral solution.

[0010] Furthermore, the terbutaline sulfate prevents and treats intestinal ischemia-reperfusion injury by reducing small intestinal pathological damage and improving the self-renewal and repair capabilities of intestinal stem cells.

[0011] The present invention also provides an application of terbutaline sulfate in preparing a medicine for preventing and treating intestinal ischemia-reperfusion injury.

[0012] Furthermore, the terbutaline sulfate is terbutaline sulfate oral solution.

[0013] The beneficial effects of the present invention are as follows: According to preliminary animal experiments and clinical sample verification, terbutaline sulfate oral solution can effectively improve the prognosis of intestinal ischemia-reperfusion injury in mice and patients. Compared with existing therapies, its advantages are:

[0014] 1. The effect is significant. Oral administration before surgery can effectively improve intestinal ischemia-reperfusion injury indicators;

[0015] 2. Small side effects, little excitatory effect on the heart, and no central nervous system effect;

[0016] 3. It is easy to use and is a commonly used drug in clinical practice. It can be aerosolized and inhaled during the perioperative period and has a rapid onset of effect.

[0017] 4. It has diverse therapeutic effects. It is a common asthma treatment drug that can improve patients’ ventilation and prognosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The experimental results of the in vitro experiment in Example 1 verifying the effect of terbutaline sulfate oral solution on the prevention and treatment of intestinal ischemia-reperfusion injury, wherein Figure A is the HE pathological score of small intestinal injury in different model mice; Figure B is the survival rate of different model mice; Figure C is the mRNA level of Lgr5 in different model mice. DETAILED DESCRIPTION

[0019] The specific product information of the terbutaline sulfate oral solution used in the present invention is as follows:

[0020] Product Name: Terbutaline Sulfate Oral Solution

[0021] Molecular formula: (C 12 H 19 NO3)2·H2SO4

[0022] Molecular weight: 548.66

[0023] Pharmacokinetics: Oral bioavailability is 15±6%, antiasthmatic effect appears in about 30 minutes. Effective blood concentration is 3μg / ml, plasma protein binding rate is 25%. The effect reaches the peak in 2 to 4 hours and lasts for 4 to 7 hours. Vd is 1.4±0.4L / kg.

[0024] Adverse reactions: Tremor, tonic spasm and palpitations are occasionally seen. The degree of adverse reactions depends on the dose and route of administration. Gradually increasing the dose from a small dose to the therapeutic dose can reduce adverse reactions. If adverse reactions occur, most of them disappear naturally within 1-2 weeks after starting medication.

[0025] Precautions: ① A few cases have finger tremor, headache, palpitations and gastrointestinal disorders. When taking 5mg orally, the incidence of finger tremor can reach 20% to 33%. ② Use with caution in patients with hyperthyroidism, coronary heart disease, hypertension, and diabetes. ③ High doses can cause ketoacidosis in patients with a history of epilepsy. ④ Long-term use can form drug resistance and reduce efficacy.

[0026] This product, terbutaline sulfate oral solution, was purchased from AstraZeneca Pharmaceuticals Co., Ltd., and the company is responsible for the specific production process. Before the occurrence of high-risk factors for intestinal ischemia-reperfusion injury, this solution should be taken orally three times a day, 10 ml each time for adults and 0.065 mg (0.20 ml) per kilogram of body weight each time for children. After obtaining informed consent from the patients, CPB patients were enrolled, and 1 ml of blood was collected from the patients 1 hour before CPB surgery and one day after surgery. After centrifugation at 10,000 rpm, 4°C, and 10 minutes, the upper plasma was drawn, and the levels of citrulline and IFABP in the patient's serum were detected according to the instructions of the citrulline kit and the IFABP kit.

[0027] Example 1 In vivo verification of terbutaline sulfate oral solution to alleviate intestinal ischemia-reperfusion injury

[0028] At the animal level, the intestinal ischemia-reperfusion injury (I / R) model was established using C57BL / 6J mice by pretreatment with different concentrations of terbutaline sulfate oral solution, and the survival rate of the mice was detected; the HE pathological damage of intestinal tissue, intestinal barrier permeability, and serum endotoxin (LPS) levels confirmed that terbutaline sulfate oral solution can alleviate intestinal ischemia-reperfusion injury (I / R).

[0029] 1. Establishment of intestinal ischemia-reperfusion injury (I / R) model: fasting for 12 hours before intestinal I / R, free drinking water, general anesthesia induced by inhalation of 4% isoflurane in C57BL / 6J mice, and then the isoflurane anesthetic dosage was maintained at 1.5% during the whole operation. The mice were fixed on the operating table in supine position, the abdominal surgical area was shaved and the skin was prepared, and after disinfection with 75% ethanol, the abdomen was opened through the midline of the abdomen, the incision was about 1-3cm, and the abdominal wall layers were cut open layer by layer to enter the abdominal cavity. The superior mesenteric artery was separated and clamped with a non-invasive micro-artery clamp to block the blood flow. After intestinal ischemia lasted for 1 hour, the abdominal cavity was opened again, the artery clamp was released to restore blood supply, and the superior mesenteric artery was reperfused. After checking that there was no bleeding in the abdominal cavity, the peritoneum, muscle and skin were sutured layer by layer with sterile silk thread. After blocking and during reperfusion, 0.5ml of warm saline at about 37°C was injected subcutaneously for fluid resuscitation. After 2 hours of reperfusion, the inferior vena cava blood, jejunum, ileum, cecum, and colon tissues were taken for examination. The sham group (sham) underwent the same I / R operation as the I / R group except that artery clamping and other I / R operations were not performed.

[0030] 2. Experimental groups:

[0031] Sixteen C57BL / 6J mice aged 6-8 weeks were randomly divided into sham operation group (Sham), intestinal ischemia-reperfusion injury model group (I / R), terbutaline sulfate oral solution + intestinal ischemia-reperfusion injury model group (TSOS+I / R), and terbutaline sulfate oral solution + sham operation group (TSOS+Sham).

[0032] (1) Sham group: except for not performing mesenteric clamping to block blood supply and removing the arterial clamp for reperfusion, all other operation steps of the mice were consistent with I / R;

[0033] (2) I / R group: intestinal I / R model was established in mice;

[0034] (3) Terbutaline sulfate oral solution + intestinal ischemia-reperfusion injury model group (TSOS+I / R): Mice were gavaged with 0.05 mg / kg terbutaline sulfate oral solution every day for 4 consecutive days, and then the intestinal I / R model was established;

[0035] (4) Terbutaline sulfate oral solution + sham operation group (TSOS+Sham): Mice were gavaged with 0.05 mg / kg of terbutaline sulfate oral solution every day for 4 consecutive days. Except for not performing mesenteric clamping to block blood supply and removing the arterial clamp for reperfusion, all other operation steps for mice were consistent with I / R.

[0036] 3. Detection indicators and technical methods:

[0037] (1) Observe and record the survival status of mice for 7 days.

[0038] (2) Evaluation of intestinal tissue damage: a. Hematoxylin and eosin (H&E) stained small intestinal segments, observed the morphological changes of small intestinal tissue under a light microscope and took photos. A pathologist who was unaware of the experimental grouping evaluated the tissue damage according to the intestinal injury scoring standard. b. Intestinal barrier permeability detection: 1) Fluorescein isothiocyanate-labeled dextran (FITC-dextran) was used to detect intestinal permeability: mice were gavaged at a dose of 0.6 mg / kg 4 hours before the test, and the FITC content in peripheral blood was detected using an enzyme marker (485 nm, 530 nm); 2) Detection of endotoxin (LPS) levels in plasma: Detection was performed according to the steps provided in the ELISA kit instructions. c. RT-PCR was used to detect the mRNA expression level of inflammatory factors in intestinal tissue.

[0039] 4. Experimental results:

[0040] As shown in Figure A below, the intestinal IR model in mice significantly aggravated the HE pathological score of small intestinal injury, while terbutaline treatment significantly reduced the small intestinal pathological injury score; in addition, the survival rate of intestinal I / R mice was significantly improved after terbutaline treatment (Figure B), and the mRNA level of Lgr5, a marker of intestinal stem cell self-renewal and repair ability, was significantly increased (Figure C). These results show that terbutaline can significantly reduce intestinal I / R injury and is a potential drug for the prevention and treatment of intestinal I / R injury.

[0041] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be covered by the scope of the claims of the present invention.

Claims

1. The use of terbutaline sulfate in the preparation of a medicament for preventing and treating intestinal ischemia-reperfusion injury, characterized in that: The terbutaline sulfate prevents and treats intestinal ischemia-reperfusion injury by reducing small intestinal pathological damage and improving the self-renewal and repair capabilities of intestinal stem cells; The terbutaline sulfate is terbutaline sulfate oral solution.

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