阅读说明：本技术 罗替戈汀制备抗急性肝损伤药物的新应用 (New application of rotigotine in preparation of medicine for resisting acute liver injury ) 是由 岳淑敏 王天 杨韵琦 于 2020-12-22 设计创作，主要内容包括：本发明涉及药学领域,本发明公开了罗替戈汀制备抗急性肝损伤药物的新应用,具体公开了罗替戈汀用于制备抗急性肝损伤药物的用途。罗替戈汀可以降低脂多糖(LPS)/D-氨基半乳糖(D-Gal)诱导的肝损伤小鼠血清谷丙转氨酶(ALT)和谷草转氨酶(AST)水平,降低急性肝损伤小鼠血清TNF-α浓度,减轻肝组织病理损伤,减少肝组织弥漫性出血。本发明的药理实验证明罗替戈汀具有减轻急性肝损伤作用,为肝损伤提供新的预防或治疗药物。(The invention relates to the field of pharmacy, discloses a new application of rotigotine in preparing an anti-acute liver injury medicament, and particularly discloses an application of rotigotine in preparing an anti-acute liver injury medicament. Rotigotine can reduce levels of Lipopolysaccharide (LPS)/D-galactosamine (D-Gal) induced liver injury mouse serum alanine Aminotransferase (ALT) and glutamic oxaloacetic transaminase (AST), reduce concentration of acute liver injury mouse serum TNF-alpha, alleviate pathological injury of liver tissue, and reduce liver tissue diffuse hemorrhage. Pharmacological experiments prove that the rotigotine has the effect of relieving acute liver injury, and provides a new preventive or therapeutic drug for liver injury.)

1. Application of rotigotine in preparing medicine for treating acute liver injury is provided.

2. The use according to claim 1, characterized in that the acute liver injury is LPS/D-Gal induced liver injury.

3. Use according to claim 1, characterized in that the subcutaneous injection of rotigotine prevents acute liver injury.

Technical Field

The invention relates to the field of pharmacy, in particular to a new application of rotigotine in preparing an anti-acute liver injury medicament.

Background

Liver damage refers primarily to damage of liver tissue. Liver damage can be caused by a variety of factors, such as viral infection, excessive intake of ethanol, improper medication, exposure to chemical poisons, ischemia reperfusion, radiation damage, and the like. Acute liver injury refers to the rapid occurrence of hepatocyte injury within a short period of time, leading to abnormal liver function. Failure to effectively control severe or persistent liver damage in a timely manner ultimately leads to liver failure. Various inflammatory mediators and proinflammatory cytokines released during the development of liver injury can also cause systemic inflammatory reactions, which further involve extra-hepatic organs.

Rotigotine is a non-ergot dopamine receptor agonist, improves tremor and dyskinesia symptoms by acting on dopamine D3/D2/D1 receptors, reduces muscle tension and plays a role in simulating dopamine. Rotigotine is mainly used for treating early-stage Parkinson's disease, can effectively relieve symptoms of the Parkinson's disease, and can also prevent the development of the Parkinson's disease to a great extent. Rotigotine is used as a Parkinson disease treatment drug, and the action mechanism of rotigotine is related to the stimulation effect of rotigotine on dopamine receptors in the tail putamen of the brain (the tail putamen region of the brain has the function of regulating movement). Rotigotine can also treat moderate to severe primary Restless Leg Syndrome (RLS). Pharmacological research also finds that rotigotine has a stimulating effect on 5-hydroxytryptamine receptors, so that an anxiolytic effect of the isobutanospiroketone is generated. In vitro multiple functional experimental studies of mice find that rotigotine has an inhibitory effect on mice vas deferens stimulated convulsion, and the inhibitory effect of rotigotine is obviously stronger than that of apomorphine, quinpirole, bromocriptine and the like. Published pharmacological studies show that: rotigotine has antiparkinsonian effect and neuroprotective effect.

However, the prior art does not disclose any information that rotigotine has anti-acute liver injury properties.

Disclosure of Invention

The invention aims to provide a novel application of rotigotine in preparing an anti-acute liver injury medicament, thereby providing a novel preventive and therapeutic medicament for liver injury.

In order to achieve the purpose, the invention adopts the technical scheme that:

application of rotigotine in preparing medicine for treating acute liver injury is provided.

Preferably, the acute liver injury is LPS/D-Gal induced liver injury.

Preferably, the rotigotine has the effect of resisting acute liver injury after subcutaneous injection.

The medicine for resisting acute liver injury prepared by rotigotine provided by the invention can relieve pathological injury of liver tissues. Also, the treatment method includes, but is not limited to, the steps of: rotigotine is administered to the drug-affected subject prior to the subject undergoing liver damage.

The medicine for resisting acute liver injury prepared by rotigotine provided by the invention can relieve pathological injury of liver tissues. The analysis reasons are shown in pharmacological experiments of the invention that rotigotine can reduce serum glutamic-pyruvic transaminase (ALT) and glutamic-oxaloacetic transaminase (AST) levels of mice with liver injury induced by Lipopolysaccharide (LPS)/D-galactosamine (D-Gal), reduce serum TNF-alpha concentration of mice with acute liver injury, relieve pathological injury of liver tissues and reduce diffuse hemorrhage of the liver tissues.

Has the advantages that:

the invention provides a new application of rotigotine in preparing an anti-acute liver injury medicament, and the application comprises the following steps: the invention provides a new prevention idea and a new treatment medicine for acute liver injury, and relates to application of rotigotine in preventing acute liver injury, a liver injury treatment medicine and a liver injury treatment method.

Drawings

The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention.

FIG. 1: effect of rotigotine on LPS/D-Gal induced pathological damage of liver tissue in mice with acute liver injury (400-fold magnified photographs). Wherein A is a control group; b is a model group; c is a rotigotine 0.35 mg/kg dose group; d is a rotigotine 0.7 mg/kg dose group; e is a rotigotine 1.4 mg/kg dose group.

FIG. 2: the rotigotine has the inhibition effect on the serum AST increase of mice with acute liver injury induced by LPS/D-Gal;

FIG. 3: the rotigotine has the inhibition effect on the serum ALT (ALT level) rise of mice with acute liver injury induced by LPS/D-Gal;

FIG. 4: effect of rotigotine on serum TNF- α levels in LPS/D-Gal induced acute liver injury mice.

Detailed Description

For a better understanding of the present disclosure, reference is made to the following detailed description of the present disclosure in connection with the specific embodiments thereof. Before the present detailed description is further described, it is to be understood that the invention is not limited to the following examples. The present invention will now be described in more detail with reference to the accompanying drawings, in which preferred embodiments of the invention are shown, it being understood that one skilled in the art may modify the invention herein described while still achieving the beneficial results of the present invention. Accordingly, the following description is to be construed as broadly as the skilled person will be able to, and not as a limitation on, the present invention.

Example 1 experiment of LPS/D-Gal induced acute liver injury in mice

Materials and reagents

Rotigotine (Sigma-Aldrich, USA); lipopolysaccharide (Sigma-Aldrich, USA); d-galactosamine (Sigma-Aldrich, USA); alanine Aminotransferase (ALT) test kit (Nanjing institute for bioengineering); aspartate Aminotransferase (AST) test kit (Nanjing institute of bioengineering); mouse tumor necrosis factor-alpha (TNF-alpha) kit (Beijing Soilebao Co., Ltd.).

Animals: SPF-grade BALB/c mice, male, 18-22 g in weight, provided by Jinanpunyue laboratory animal breeding Co., Ltd, and the qualification number: SCXK (lu) 20190003.

The instrument comprises the following steps: electronic balance (Shanghai Mettler Optiphot Corp); low temperature centrifuge (shanghai Eppendorf corporation); an incubator (Shanghai Long-flow scientific instruments, Inc.); SpectraMax M3 multifunctional microplate reader (Molecular Devices, USA).

1) The experimental steps of the LPS/D-Gal induced acute liver injury of the mice are as follows:

BALB/c male mice (18-22 g) were 40 mice, and were randomly divided into 8 mice per group, a control group, a model group, a rotigotine 0.35 mg/kg dose group, a rotigotine 0.7 mg/kg dose group, and a rotigotine 1.4 mg/kg dose group. After 3 days of adaptive feeding, mice in a control group are injected with 1% of tween 20 subcutaneously, mice in a model group are injected with 1% of tween 20 subcutaneously, and rotigotine is injected subcutaneously according to respective administration doses in groups of 0.35, 0.7 and 1.4 mg/kg of rotigotine. After 2 hours, the control mice were injected with 0.9% sodium chloride injection. Model group, rotigotine 0.35 mg/kg dose group, rotigotine 0.7 mg/kg dose group, rotigotine 1.4 mg/kg dose group mice were injected i.p. with LPS (30. mu.g/kg)/D-Gal (800 mg/kg). After 6 hours from the LPS/D-Gal injection, blood and liver tissues of the mice were collected.

2) Pathological observation of liver tissue: 6 hours after LPS/D-Gal injection, 3 mice were randomly selected per group, euthanized with excess chloral hydrate, and the right lobe liver of the mice (this was done on ice) was placed in 4% paraformaldehyde tissue fixative overnight. The solution is dehydrated in a glass jar by alcohol gradient, and is sliced after paraffin embedding. Hematoxylin-eosin staining, sealing with neutral resin, and observing pathological changes of liver tissue under optical microscope.

As shown in fig. 1, the histopathological results show that: 1. the liver lobule structure of the liver tissue of the control group mouse is clear, the liver cells are regularly arranged, and the structure and the shape of the liver cells are normal. 2. The liver tissue of the mouse in the model group is subjected to diffuse bleeding, the arrangement of liver cells is disordered, and a large amount of inflammatory cell infiltration, large-scale hepatocyte necrosis and cell nucleus lysis can be seen around the central vein. 3. The liver tissue bleeding of mice in the rotigotine dose groups of 0.35, 0.7 and 1.4 mg/kg is reduced, the liver cells are relatively orderly arranged, the inflammatory cell infiltration is reduced, and the liver cell necrosis is obviously reduced.

Among them, FIG. 1 (photograph after magnification of 400 times) shows that the LPS/D-Gal-induced liver tissue damage of mice can be reduced by injecting rotigotine before injecting LPS/D-Gal into the mice. The invention provides a new application of rotigotine in preparing a medicament for preventing acute liver injury. The rotigotine provided by the invention can be used as a drug for preventing acute liver injury, and is characterized in that the rotigotine reduces the serum AST and ALT rise of mice with acute liver injury induced by LPS/D-Gal, reduces the serum TNF-alpha level, and relieves the pathological injury of liver tissues, so that the rotigotine has the effect of relieving the liver injury.

The invention further proves the analysis through the concentration detection experiment of the serum AST and ALT of the mouse with acute liver injury induced by LPS/D-Gal and the level detection experiment of the serum TNF-alpha of the mouse after liver injury.

Example 2

1) And (3) detecting the AST and ALT activities of the serum: after 6 hours of LPS/D-Gal injection, the mice of example 1 were bled from the canthus veins of the eyes, left at room temperature for 0.5 hours, at 4 ℃ 3500 r/min and centrifuged for 15 min. The supernatant was pipetted into a new EP tube. Serum AST and ALT activities were measured using alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) test kits, respectively. Results are expressed as mean ± standard deviation (n = 8).

As shown in FIGS. 2 and 3, the serum AST and ALT activities of the mice in the model group were significantly increased compared with those in the control group (^** P<0.01); serum AST Activity of mice in the rotigotine 0.35 mg/kg dose group was reduced compared to the model group (^# P<0.05); compared with the model group, the serum AST and ALT activities of mice in the rotigotine 0.7 and 1.4 mg/kg dose group are obviously reduced (^## P<0.01）。

The results in FIG. 2 and FIG. 3 show that the injection of rotigotine before the injection of LPS/D-Gal in mice can reduce the activity of AST and ALT in mice with acute liver injury induced by LPS/D-Gal.

2) Serum TNF-alpha concentration detection: after 6 hours of LPS/D-Gal injection, the mice of example 1 were bled from the canthus veins of the eyes, left at room temperature for 0.5 hours, at 4 ℃ 3500 r/min and centrifuged for 15 min. The supernatant was pipetted into a new EP tube. Mouse serum TNF-alpha levels were detected using a mouse tumor necrosis factor-alpha (TNF-alpha) kit. Results are expressed as mean ± standard deviation (n = 8).

The detection result is shown in figure 4, compared with the control group, the serum TNF-alpha level of the mouse in the model group is obviously increased (^** P<0.01); compared with the model group, the serum TNF-alpha level of mice in the rotigotine dosage groups of 0.35, 0.7 and 1.4 mg/kg is obviously reduced (^## P<0.01）。

The results in FIG. 4 show that the serum TNF-alpha level of mice with acute liver injury induced by LPS/D-Gal can be reduced by injecting rotigotine before the mice are injected with LPS/D-Gal.

The key of the effect of rotigotine on acute liver injury is to reduce the increase of serum AST and ALT of mice with acute liver injury induced by LPS/D-Gal by rotigotine, reduce the level of serum TNF-alpha and relieve the pathological injury of liver tissues by detecting the serum AST and ALT concentration of mice with acute liver injury induced by LPS/D-Gal, observing the pathological observation of liver tissues and detecting the level of serum TNF-alpha after the liver injury of the mice.

Although the preferred embodiments of the present invention have been described in detail, the present invention is not limited to the details of the embodiments, and various equivalent modifications can be made within the technical spirit of the present invention, and the scope of the present invention is also within the scope of the present invention. It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition. In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.