Application of genkwanin in preparation of anti-depression drugs

Abstract

The invention belongs to the technical field of medicines, and discloses application of genkwanin in preparation of an anti-depression medicine. The pharmaceutical application of genkwanin is expanded, the anti-depression effect of genkwanin is researched by analyzing the influence of genkwanin on mouse depression-like behaviors, and it is proved that genkwanin can remarkably shorten the time of forced swimming and tail suspension immobility of chronic depression model mice; the time of forced swimming and tail suspension immobility of acute depression model mice can be remarkably shortened, and the pharmaceutical composition has a remarkable anti-depression effect and is small in dosage; the composition has no influence on heart, liver, spleen, lung and kidney of acute and chronic depression model mice, is safe and has no toxic or side effect. The genkwanin is applied to the preparation of the antidepressant drug, has a good application value, and provides a new drug application for the genkwanin.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of genkwain in the preparation of antidepressant drugs. Background Art

[0002] Depression is a multiphasic disorder, including emotional, cognitive, behavioral and somatic regulation disorders. Long-term stress and acute inflammation are very important factors in inducing depression. However, common antidepressants currently used in clinical practice have problems such as slow onset, many adverse reactions, low efficacy and low selectivity. Therefore, the continuous exploration of new antidepressants has important application value. At present, researchers at home and abroad are gradually focusing on extracting safer and more effective antidepressants from natural plants.

[0003] Lobelia chinensis is a traditional Chinese medicine for clearing away heat and detoxifying. It comes from the dried whole herb of Lobelia chinensis, a plant of the Campanulaceae family. The Compendium of Materia Medica records that Lobelia chinensis has a pungent, flat, non-toxic smell and is mainly used to treat snake bites, asthma, and malaria. According to a document released by the National Health Commission, Lobelia chinensis can be used as both medicine and food within a limited range of use and dosage. Modern research has found that Lobelia chinensis has pharmacological effects such as diuresis, antibacterial, choleretic, and anti-tumor. Genkwanin is a monomer component extracted from Lobelia chinensis and is a flavonoid compound. Genkwanin is widely used in drug research and development and manufacturing. At present, studies have shown that Genkwanin has antioxidant, anti-aging, anti-inflammatory, and anti-tumor effects, but there are no reports on its use in the treatment of depression. Summary of the invention

[0004] In order to obtain safer and more effective natural antidepressant drugs, the present invention expands the pharmaceutical application of Genkandrin and studies its application in the preparation of antidepressant drugs.

[0005] By analyzing the effect of genkwain on the depressive-like behavior of mice and studying its antidepressant effect, it was found that genkwain has the activity of treating depression. Therefore, the purpose of the present invention is to provide the use of genkwain in the preparation of antidepressant drugs.

[0006] To achieve the purpose of the present invention, the technical solution adopted by the present invention is as follows: the structural formula of Genkwanin is as follows:

[0007]

[0008] The present invention uses a chronic depression model induced by chronic unpredictable mild stress (CUMS) and an acute depression model induced by lipopolysaccharide (LPS) to study depression. Under chronic unpredictable mild stress or lipopolysaccharide conditions, animals develop depressive-like behaviors. The present invention found that compared with mice in the negative control group, mice in the CUMS model group showed depressive-like behaviors, and Genkwain was able to improve the depressive-like behaviors of mice induced by CUMS; compared with mice in the negative control group, the LPS model group showed depressive-like behaviors, and Genkwain was able to improve the depressive-like behaviors of mice induced by LPS. In addition, the present invention also found that compared with venlafaxine, an antidepressant with a strong antidepressant effect commonly used in clinical practice, a medicinal dose of 2 mg / kg of Genkwain can achieve the antidepressant effect of a conventional medicinal dose of 10 mg / kg of venlafaxine. Therefore, Genkwain has a significant antidepressant effect.

[0009] According to the above, the beneficial effects of the present invention are as follows:

[0010] 1) The present invention experimentally analyzes the effect of Genkwanin on the depressive-like behavior of mice, confirming that Genkwanin can significantly shorten the forced swimming and tail suspension immobility time of chronic depression model mice, and has a significant antidepressant effect; it can significantly shorten the forced swimming and tail suspension immobility time of acute depression model mice, and has a significant antidepressant effect. The application of it in the development of antidepressant drugs has great application value, and provides a new drug use for Genkwanin.

[0011] 2) Genkwanin has a small dosage as an antidepressant preparation and is derived from natural plants. It can be used as both medicine and food. It is used to prepare antidepressant drugs with small side effects and high safety. The present invention confirms that Genkwanin has no effect on the heart, liver, spleen, lung, and kidney of acute and chronic depression model mice, and has good safety.

[0012] 3) The present invention expands the types of natural antidepressant drugs and discovers the effective ingredients in Lobelia that are rich in antidepressant effects, providing new theoretical support for the expansion and effective use of Lobelia. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Effects of Genkwanin on the immobility time of forced swimming in chronic depression model mice;

[0014] Figure 2 Effect of Genkwanin on the tail suspension immobility time in chronic depression model mice;

[0015] Figure 3 Effects of Genkwanin on the immobility time of forced swimming in acute depression model mice;

[0016] Figure 4Effect of Genkwanin on the tail suspension immobility time in acute depression model mice;

[0017] Figure 5 Effects of Genkwanin on the heart, liver, spleen, lung and kidney of chronic depression model mice;

[0018] Figure 6 Effects of genkwain on the heart, liver, spleen, lung and kidney of mice with acute depression model. DETAILED DESCRIPTION

[0019] The present invention is further described in detail below with reference to the embodiments. Unless otherwise specified, the percentages are all by mass.

[0020] In order to illustrate the application of Genkwanin in the present invention in anti-depression, the present invention conducted the following animal experiment related research.

[0021] 1. Experimental Drugs

[0022] Genkwain powder was obtained from commercial sources. Genkwain powder was dissolved in 0.5% CMC-Na to prepare 0.2 mg / ml and 0.4 mg / ml dosing solutions, respectively, with a dosing volume of 0.05 ml / 10 g, and the dosing method was intragastric administration, that is, Genkwain was administered at a dose of 1 mg / kg and 2 mg / kg, respectively.

[0023] Venlafaxine was obtained from commercial sources. Venlafaxine powder was dissolved in physiological saline to prepare a 2 mg / ml dosing solution with a dosing volume of 0.05 ml / 10 g. The dosing method was intragastric administration, that is, venlafaxine was administered at a dose of 10 mg / kg.

[0024] LPS was obtained from commercial sources. LPS powder was dissolved in physiological saline to prepare a 0.05 mg / ml dosing solution with a dosing volume of 0.1 ml / 10 g. The dosing method was intraperitoneal injection, that is, LPS was administered at a dose of 0.5 mg / kg.

[0025] 2. Experimental Animals

[0026] Healthy male C57BL / 6 mice, weighing 22-24 g and aged 6-8 weeks, were purchased from Beijing Sibeifu Biotechnology Co., Ltd. The animals were allowed to eat freely and were allowed to adapt to the environment in the laboratory before the experiment.

[0027] 3. Establishment of the Mouse Chronic Depression Model and Animal Drug Administration

[0028] The present invention adopts chronic unpredictable mild stress (CUMS) to prepare a chronic depression model in mice. Normal C57BL / 6 mice were randomly divided into a negative control group and a stress modeling group, and the following multiple stimulation factors were used for modeling: ① 2h restraint; ② 3h tilting the mouse cage at 45°; ③ fasting for 24h; ④ 24h water deprivation; ⑤ alternating lighting overnight; ⑥ 3h wet bedding. After 28 days of modeling, behavioral testing was performed. The animals with successful modeling were randomly divided into a CUMS group, a CUMS+1mg / kg genkwain group, a CUMS+2mg / kg genkwain group, and a CUMS+10mg / kg venlafaxine (positive drug) group, and the modeling was continued for 14d. During the modeling period, all animals were gavage-administered once a day for 14 consecutive days. The mice in the negative control group were kept in a breeding cage.

[0029] Animal modeling and drug administration were as follows:

[0030] Negative control group: mice were not subjected to chronic stress modeling; 0.5% CMC-Na was administered by intragastric gavage at a volume of 0.05 ml / 10 g.

[0031] CUMS group: mice were subjected to chronic stress modeling; 0.5% CMC-Na was administered by oral gavage at a volume of 0.05 ml / 10 g.

[0032] CUMS+1mg / kg Genkwanin group: mice were subjected to chronic stress; 0.2mg / ml Genkwanin was administered by oral gavage at a volume of 0.05ml / 10g, i.e. 1mg Genkwanin was administered for every 1kg body weight of mice.

[0033] CUMS+2mg / kg Genkwanin group: mice were subjected to chronic stress; 0.4mg / ml Genkwanin was administered by oral gavage at a volume of 0.05ml / 10g, i.e. 2mg Genkwanin was administered for every 1kg body weight of mice.

[0034] CUMS+10mg / kg venlafaxine (positive drug) group: mice were subjected to chronic stress; 2mg / ml venlafaxine was administered by oral gavage at a volume of 0.05ml / 10g, that is, 10mg venlafaxine was administered for every 1kg body weight of mice.

[0035] 4. Establishment of Acute Depression Model in Mice and Drug Administration

[0036] The present invention adopts LPS to prepare an acute depression model in mice. Normal C57BL / 6 mice are randomly divided into a negative control group, an LPS group, an LPS+1 mg / kg genkwain group, an LPS+2 mg / kg genkwain group, and an LPS+10 mg / kg venlafaxine (positive drug) group. The mice are administered once a day for 7 consecutive days.

[0037] Animal modeling and drug administration were as follows:

[0038] Negative control group: physiological saline was intraperitoneally injected at a volume of 0.1 ml / 10 g; 0.5% CMC-Na was intragastrically administered at a volume of 0.05 ml / 10 g.

[0039] LPS group: 0.05 mg / ml LPS was intraperitoneally injected at a volume of 0.1 ml / 10 g, that is, 0.5 mg LPS was administered per 1 kg body weight of mice; 0.5% CMC-Na was intragastrically administered at a volume of 0.05 ml / 10 g.

[0040] LPS+1mg / kg Genkwanin group: 0.05mg / ml LPS was administered by intraperitoneal injection at a volume of 0.1ml / 10g, i.e., 0.5mg LPS was administered for every 1kg body weight of mice; 0.2mg / ml Genkwanin was administered by oral gavage at a volume of 0.05ml / 10g, i.e., 1mg Genkwanin was administered for every 1kg body weight of mice.

[0041] LPS+2mg / kg Genkwanin group: 0.05mg / ml LPS was administered by intraperitoneal injection at a volume of 0.1ml / 10g, i.e. 0.5mg LPS was administered for every 1kg body weight of mice; 0.4mg / ml Genkwanin was administered by oral gavage at a volume of 0.05ml / 10g, i.e. 2mg Genkwanin was administered for every 1kg body weight of mice.

[0042] LPS+10mg / kg venlafaxine (positive drug) group: 0.05mg / mlLPS was administered by intraperitoneal injection at a volume of 0.1ml / 10g, i.e., 0.5mg LPS was administered per 1kg body weight of mice; 2mg / ml venlafaxine was administered by oral gavage at a volume of 0.05ml / 10g, i.e., 10mg venlafaxine was administered per 1kg body weight of mice.

[0043] 5. Animal behavior testing

[0044] Forced swim test (FST) and tail suspension test (TST) are the most widely used test methods for screening antidepressant drugs. They are used to evaluate the degree of depression of animals by testing the state of despair (i.e., immobility) of animals after struggling under forced swimming or tail suspension stress conditions. In the forced swim test, the immobility time of animals in water is used as an indicator to evaluate the degree of depression of animals, while in the tail suspension test, the immobility time of animals while suspended is used to judge the degree of depression of animals.

[0045] (1) Forced swimming test: The mice were placed in a plexiglass bucket (16 cm in diameter) (water depth 14 cm, water temperature 24 ± 2°C) for forced swimming. Each mouse was video recorded for 6 min, and the immobility time of the mouse within the last 5 min was recorded.

[0046] (2) Tail suspension experiment: The mouse tail was taped to the tail suspension device with medical tape, so that the mouse was suspended upside down with its head about 50 cm from the table. Each mouse was video recorded for 6 min, and the immobility time of the mouse within the next 5 min was recorded.

[0047] 6. HE staining

[0048] The tissue samples were fixed, embedded, and sliced ​​in paraffin (thickness 4 μm) according to the routine procedures, and HE staining was performed. A high-resolution pathology section scanning and analysis system (3DHISTECH / Pannoramic SCAN) was used for image acquisition.

[0049] 7. Experimental Results

[0050] Effects of Genkwanin on the immobility time of forced swimming in chronic depression model mice Figure 1 As shown. Compared with the mice in the negative control group, the immobility time of mice in the CUMS model group was significantly increased (p < 0.05). Compared with the CUMS model group, the immobility time of mice in the CUMS model + 1 mg / kg genkwain group was significantly decreased (p < 0.01). Compared with the CUMS model group, the immobility time of mice in the CUMS model + 2 mg / kg genkwain group was significantly decreased (p < 0.05). The immobility time of mice in the CUMS + 10 mg / kg venlafaxine (positive drug) group was also significantly shortened compared with that of mice in the CUMS model group (p < 0.01). There was no significant difference in the immobility time between mice in the CUMS model + 2 mg / kg genkwain group and mice in the CUMS + 10 mg / kg venlafaxine (positive drug) group (p > 0.05). Genkwain has similar effects to venlafaxine, both of which can significantly reduce the forced swimming immobility time of CUMS depression model mice, and a medicinal dose of 2 mg / kg of Genkwain can achieve the efficacy of a conventional medicinal dose of venlafaxine (10 mg / kg).

[0051] Effects of Genkwanin on the tail suspension immobility time in chronic depression model mice Figure 2As shown. Compared with the mice in the negative control group, the immobility time of mice in the CUMS model group was significantly increased (p < 0.01). Compared with the CUMS model group, the immobility time of mice in the CUMS model + 1 mg / kg genkwain group was significantly decreased (p < 0.05). Compared with the CUMS model group, the immobility time of mice in the CUMS model + 2 mg / kg genkwain group was significantly decreased (p < 0.01). The immobility time of mice in the CUMS + 10 mg / kg venlafaxine (positive drug) group was also significantly shortened compared with that of mice in the CUMS model group (p < 0.01). There was no significant difference in the immobility time between mice in the CUMS model + 2 mg / kg genkwain group and mice in the CUMS + 10 mg / kg venlafaxine (positive drug) group (p > 0.05). Like venlafaxine, genkwain can significantly reduce the tail suspension immobility time of CUMS depression model mice, and a medicinal dose of 2 mg / kg of genkwain can achieve the effect of a conventional medicinal dose of venlafaxine (10 mg / kg).

[0052] Effects of Genkwanin on the immobility time of forced swimming in acute depression model mice Figure 3 As shown. Compared with the negative control group mice, the immobility time of mice in the LPS model group was significantly increased (p < 0.01). Compared with the LPS model group, the immobility time of mice in the LPS model + 1 mg / kg genkwain group was significantly reduced (p < 0.01). Compared with the LPS model group, the immobility time of mice in the LPS model + 2 mg / kg genkwain group was significantly reduced (p < 0.01). Compared with the LPS model group mice, the immobility time of mice in the LPS + 10 mg / kg venlafaxine (positive drug) group was also significantly shortened (p < 0.01). There was no significant difference in the immobility time between the LPS model + 2 mg / kg genkwain group and the LPS + 10 mg / kg venlafaxine (positive drug) group (p > 0.05). Like venlafaxine, genkwain can significantly reduce the tail suspension immobility time of LPS depression model mice, and the medicinal dose of 2 mg / kg genkwain can achieve the effect of the conventional medicinal dose of 10 mg / kg venlafaxine).

[0053] Effects of Genkwanin on the tail suspension immobility time in acute depression model mice Figure 4As shown. Compared with the negative control group mice, the immobility time of mice in the LPS model group was significantly increased (p < 0.01). Compared with the LPS model group, the immobility time of mice in the LPS model + 1 mg / kg genkwain group was significantly reduced (p < 0.01). Compared with the LPS model group, the immobility time of mice in the LPS model + 2 mg / kg genkwain group was significantly reduced (p < 0.01). Compared with the LPS model group mice, the immobility time of mice in the LPS + 10 mg / kg venlafaxine (positive drug) group was also significantly shortened (p < 0.05). There was no significant difference in the immobility time between the LPS model + 2 mg / kg genkwain group and the LPS + 10 mg / kg venlafaxine (positive drug) group (p > 0.05). Genkwain has similar effects to venlafaxine, both of which can significantly reduce the immobility time of tail suspension in LPS depression model mice, and the medicinal dose of 2 mg / kg genkwain can achieve the effect of the conventional medicinal dose of 10 mg / kg venlafaxine).

[0054] Effects of Genkwanin on the heart, liver, spleen, lung and kidney of chronic depression model mice Figure 5 As shown. There were no significant changes in the structure and cell morphology of the heart, liver, spleen, lung, and kidney in the negative control group, CUMS model group, CUMS model + 1 mg / kg Genkwanin group, and CUMS model + 2 mg / kg Genkwanin group. Genkwanin had no effect on the heart, liver, spleen, lung, and kidney of CUMS depression model mice, and had good safety.

[0055] Effects of Genkwanin on the heart, liver, spleen, lung and kidney of mice with acute depression model Figure 6 As shown. There were no significant changes in the structure and cell morphology of the heart, liver, spleen, lung, and kidney in the negative control group, LPS model group, LPS model + 1 mg / kg genkwain group, and LPS model + 2 mg / kg genkwain group. Genkwain had no effect on the heart, liver, spleen, lung, and kidney of CUMS depression model mice, and had good safety.

[0056] The above experiments confirmed that developing Genkwain as an antidepressant drug has great application value.

Claims

1. The use of Genkwanin in drug preparation, characterized in that: As an active ingredient, it is used to prepare drugs for treating or preventing depression.

2. The use of Genkwanin in drug preparation according to claim 1, characterized in that: The medicinal dosage of Genkandraefolin is 2 mg / kg.

3. The use of Genkwanin in drug preparation according to claim 1 or 2, characterized in that: It is used as an active ingredient or is prepared into an oral preparation, lyophilized powder or injection preparation with pharmaceutically acceptable excipients.

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