Application of isoliensinine in preparation of anti-depression drugs

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of isoliensinine in preparation of an anti-depression medicine. Researches find that isoliensinine can significantly improve mouse depression symptoms caused by CUMS induction by relieving intracerebral inflammatory response and inhibiting excessive activation of an HPA axis, and has statistical significance. The isoliensinine is used as an alkaloid active component extracted from the lotus plumule, shows an obvious anti-depression effect and can be used for preparing medicines for preventing or treating depression. The invention provides a new therapeutic drug choice for the treatment of depression, and meanwhile, the new medicinal value of the isoliensinine can be excavated.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and relates to the application of isoliensinine, and in particular to the application of isoliensinine in the preparation of antidepressant drugs. Background Art

[0002] Depression is a common mental illness, and its clinical manifestations include persistent low mood, loss of interest, lack of pleasure, despair, etc. This emotional disorder has a profound impact on the patient's mental health and quality of life, and is accompanied by a series of symptoms such as cognitive dysfunction, sleep disorders, changes in appetite, and decreased self-worth. In some cases, patients with depression may even have suicidal thoughts or behaviors, which makes depression one of the focuses of global public health. Epidemiological studies have shown that the prevalence of depression is high worldwide, and the prevalence varies in different countries and regions, but generally affects about 3% to 5% of the world's population. The pathogenesis of depression has not yet been fully clarified, but it is generally believed to involve multiple aspects such as genetics, biochemistry, neuroendocrine, and neuroimmunity. Abnormal levels of monoamine neurotransmitters, abnormalities in neurotrophic factors, neuroinflammation, hyperactivity of the hypothalamic-pituitary-adrenal axis (HPA axis), and changes in brain structure and function are all considered to be related to the pathogenesis of depression. At present, the main methods of clinical treatment of depression are drug therapy, psychotherapy, and physical therapy. Most of the drugs used clinically to treat depression are based on the monoaminergic hypothesis, which improves symptoms by regulating the levels of serotonin, norepinephrine, and dopamine. Although these drugs are effective to a certain extent, they have obvious limitations: they are slow to take effect, usually taking several weeks to take effect; they have limited efficacy, and about one-third of patients do not respond to such drug treatments; they have many side effects, including nausea, insomnia, sexual dysfunction, etc., which affect the quality of life of patients and have poor treatment compliance. Therefore, it has become an urgent need in the field of depression treatment to develop new and effective antidepressant drugs based on the multiple mechanisms of action of depression in order to improve efficacy and reduce side effects.

[0003] The causes of depression are complex, and among the many pathogenic factors, neuroinflammation has become a key link. A large number of existing studies have confirmed that abnormal activation of microglia in the prefrontal temporal cortex and hippocampus, which are closely linked to inflammation, is one of the important causes of depression. As a key inflammatory factor, interleukin 1β (IL-1β) plays an important role in the occurrence and development of depression. Its level changes in the brain show a significant positive correlation with the severity of depressive symptoms. HPA axis hyperactivity is closely related to the occurrence and development of depression. When the body is stimulated by stress, the HPA axis is activated to promote the release of glucocorticoids. The short-term increase in glucocorticoid levels can inhibit the HPA axis through negative feedback and maintain endocrine balance. However, under long-term stress, glucocorticoids are continuously released, causing the HPA axis to be hyperactive for a long time, leading to dysfunction of glucocorticoid receptors and disordered negative feedback mechanisms of the HPA axis, which in turn induces or aggravates depression.

[0004] Isoliensinine (abbreviation: IL) is a dibenzylisoquinoline alkaloid extracted from the green embryo (lotus seed heart) of the mature seeds of the plant Nelumbonucifera Gaertn. of the Nymphaeaceae family. Studies have shown that isoliensinine has multiple pharmacological effects such as antihypertensive, antiarrhythmic, anti-inflammatory, anti-tumor and anti-fibrosis. However, there are no reports on whether isoliensinine can prevent and treat depression by inhibiting neuroinflammation and HPA axis hyperactivity in the brain. Summary of the invention

[0005] In view of the problems existing in the prior art, the present invention aims to provide an application of isoliensinine in the preparation of antidepressant drugs.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] Application of isoliensinine in the preparation of antidepressant drugs.

[0008] Further, in the above application, the structural formula of isoliensinine is shown as (I):

[0009]

[0010] Furthermore, in the above application, the anti-depressant effects include improving depressive behavior, reducing inflammatory response in the brain, and inhibiting hyperactivity of the HPA axis.

[0011] Furthermore, in the above application, the dosage form of the drug is any dosage form acceptable in pharmacological therapy.

[0012] Furthermore, in the above application, the dosage of the drug is any therapeutically acceptable dosage.

[0013] Compared with the prior art, the present invention has the following beneficial effects:

[0014] 1. The present invention explores the new medicinal value of isoliensinine, and its application in the preparation of drugs for preventing or treating depression provides a new drug option for the prevention and treatment of depression.

[0015] 2. The present invention has found through research that isoliensinine can significantly improve the depressive behavior of chronic unpredictable mild stress (CUMS) model mice, with statistical significance. And isoliensinine can play an antidepressant effect by alleviating the inflammatory response in the mouse brain and inhibiting the hyperactivity of the HPA axis, and can be used to prepare drugs for preventing or treating depression. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The effect of isoliensinine on the sugar preference index in the sugar preference experiment of CUMS depressed mice. Compared with the CUMS group, * P<0.05, ** P<0.01, n=9.

[0017] Figure 2 The effect of isoliensinine on the immobility time of the tail suspension test in CUMS depressed mice. ** P<0.01, *** P<0.001, n=9.

[0018] Figure 3 The effect of isoliensinine on the number of grid crossings (a) and central residence time (b) of CUMS depressed mice in the open field test. ns P>0.05, * P<0.05, ** P<0.01, n=9.

[0019] Figure 4 Effects of isoliensinine on IL-1β levels in the hippocampus (a) and cortex (b) of CUMS-depressed mice. * P<0.05, ** P<0.01, *** P<0.001, n=3.

[0020] Figure 5 The effect of isoliensinine on the expression of serum corticosterone (CORT) in CUMS depressed mice. ns P ≥ 0.05, ** P<0.01, *** P<0.001, n=6. DETAILED DESCRIPTION

[0021] The technical scheme and technical effects of the present invention are described in detail below in conjunction with specific embodiments and drawings. Experimental methods without specifying specific conditions are usually based on conventional conditions, such as the conditions described in textbooks and experimental guides, or the conditions recommended by manufacturers, which are well known or easily known to ordinary technicians in the field. The following embodiments are only preferred embodiments of the present invention and do not limit the present invention. For those skilled in the art, the present invention can have various changes and variations. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

[0022] Example 1

[0023] The isoliensinine (IL; purity>99%) used in the examples was purchased from Munster Biotechnology Co., Ltd., and the structural formula is shown below:

[0024]

[0025] 1. Experimental methods

[0026] 1. Preparation of chronic unpredictable mild stress (CUMS) depression mouse model

[0027] SPF male Kunming mice (KM) aged 6-8 weeks and weighing 18-22g were selected. After one week of adaptive feeding, the following stimulations were performed: water deprivation for 12h, food deprivation for 12h, long day, wet bedding for 24h, bedding deprivation for 24h, noise stimulation for 2h, LED flashing for 2h, tail clamping for 2min, shaking the mouse cage for 2h, swimming in 4℃ ice water for 5min, tilting the cage for 12h, and exchanging the same cage for 12h. 2-3 kinds of stimulations were randomly selected from the above stimulations every day, and they were not repeated for two consecutive days. After 7 weeks, depressive behavioral tests were performed to determine whether the CUMS model was successfully established.

[0028] 2. Experimental Grouping

[0029] 45 KM mice were randomly divided into 5 groups, 9 mice in each group, namely blank control group (CON), CUMS group, CUMS+5mg / kg IL group, CUMS+10mg / kg IL group and CUMS+10mg / kg FLX (Fluoxetine) group. The mice in CUMS, CUMS+5mg / kg IL, CUMS+10mg / kg IL and CUMS+10mg / kg FLX groups were subjected to CUMS for 7 weeks to establish the depression mouse model (the mice in CON group were not subjected to CUMS). The 5 groups of mice were treated with medication for 3 weeks starting from the 5th week, that is, IL (5 and 10 mg / kg) or FLX (10 mg / kg) was intraperitoneally injected every day according to different groups (CUMS+5mg / kg IL, CUMS+10mg / kg IL and CUMS+10mg / kg FLX groups), and the CON group and CUMS group were intraperitoneally injected with the same volume of sterile saline. Behavioral experiments were performed on the day after the last administration.

[0030] 3. Depression-related behavioral experiments

[0031] (1) Sugar water preference test (SPT): On the first day, mice were provided with two bottles of 1% (w / v) sucrose solution; on the second day, one bottle of sugar water was replaced with ordinary water; on the third day, mice were deprived of water and food; on the fourth day, a pre-weighed bottle of sugar water and a bottle of ordinary water were placed in the cage at the same time for 24 hours (the positions were changed every 12 hours to avoid position preference). After the experiment, the sugar water bottle and the ordinary water bottle were weighed to calculate the sugar water preference index. Sugar water preference index (%) = sugar water consumption / (ordinary water consumption + sugar water consumption) × 100%.

[0032] (2) Tail suspension test (TST): Use medical tape to wrap a circle around the mouse's tail 1 cm away from the tip of the tail. Then use a clip to clamp the tape instead of clamping the mouse's tail directly to avoid causing pain. The mouse is suspended at a high place, ensuring that the mouse's head is at least 15 cm from the ground. Cardboard is used to block the mouse's vision to reduce external interference. The experiment lasts for 6 minutes, of which the first 2 minutes is the adaptation period and no data is recorded. The last 4 minutes record the length of time the mouse is immobile (the mouse is considered to be in an immobile state if its limbs do not make any obvious and violent movements while hanging).

[0033] (3) Open field test (OFT): Place the mice in an opaque open field box with a length, width, and height of 50 cm, 50 cm, and 40 cm respectively. Draw a 5×5 grid on the bottom, with the 9 grids in the middle as the center area. Allow the mice to move freely in the open field box for 5 minutes. Record and analyze the number of grid crossings and the time the mice stay in the center area within 5 minutes to evaluate their spontaneous activity and anxiety. Before each mouse is placed in the open field box, wipe the open field box with 75% alcohol to avoid interference from the smell of the previous mouse.

[0034] 4. Enzyme-linked immunosorbent assay

[0035] Method for obtaining hippocampal and cortical tissue samples: Three mice were randomly selected from each group and killed by cervical dislocation. The hippocampus and cortex in the brain were separated and homogenized in sample lysis buffer, centrifuged at 4°C to obtain the supernatant. Method for obtaining blood samples: Six mice were randomly selected from each group, and blood was collected from the heart after intraperitoneal injection of 4% sodium pentobarbital (0.025mL / 10g) for anesthesia. After standing at room temperature for 2 hours, the supernatant was obtained by centrifugation at 4°C. The levels of IL-1β and CORT in the samples were detected using IL-1β and corticosterone (CORT) ELISA kits (Andy Gene, Beijing, China) according to the standard operating procedures in the kit instructions.

[0036] (II) Experimental results

[0037] 1. Isoliensinine increases the sugar preference index of CUMS depressed mice

[0038] In the sugar water preference experiment, the sugar water preference index of mice reflects the anhedonia of mice. Figure 1 As shown in the figure, compared with the CON group, the sugar preference index of mice in the CUMS group was significantly reduced (P < 0.01). Compared with the CUMS group, the sugar preference index of mice in the drug group was significantly increased after IL and FLX treatment (IL 5mg / kg: P < 0.05; IL 10mg / kg: P < 0.01; FLX 10mg / kg: P < 0.05). This shows that isoliensinine can significantly reduce the degree of anhedonia in CUMS depressed mice.

[0039] 2. Isoliensinine reduces the tail suspension immobility time of CUMS depressed mice

[0040] In the tail suspension test, the immobility time of mice after giving up struggling represents the despair of mice and can reflect their degree of depression. Figure 2As shown in the figure, compared with the CON group, the immobility time of mice in the CUMS group was significantly increased (P < 0.01). Compared with the CUMS group, the immobility time of mice in the drug administration group was significantly reduced after IL and FLX treatment (IL 5mg / kg: P < 0.05; IL 10mg / kg: P < 0.01; FLX 10mg / kg: P < 0.05). This shows that the administration of isoliensinine significantly inhibited the behavioral despair of CUMS depressed mice.

[0041] 3. Isoliensinine increases the number of grid crossings and central residence time of CUMS depressed mice in the open field test

[0042] In the open field test, the number of grid crossings of mice can be used to understand the spontaneous movement and anxiety level of mice. Figure 3 As shown in a, compared with the CON group, the number of grid crossings of mice in the CUMS group was significantly reduced (P < 0.01). Compared with the CUMS group, the immobility time of mice in the drug-treated group was significantly reduced after IL and FLX treatment (IL 5mg / kg: P < 0.05; IL 10mg / kg: P < 0.01; FLX10mg / kg: P < 0.01), indicating that the administration of isoliensinine can significantly increase the spontaneous movement behavior of CUMS depressed mice. Figure 3 As shown in b, compared with the CON group, the central residence time of mice in the CUMS group was significantly reduced (P < 0.01). Compared with the CUMS group, the central residence time of mice in the drug group was significantly increased after IL and FLX treatment (IL 10 mg / kg: P < 0.01; FLX 10 mg / kg: P < 0.01), indicating that isoliensinine effectively alleviated the anxiety state of CUMS depressed mice.

[0043] 4. Isoliensinine reduces the level of inflammatory factor IL-1β in the hippocampus and cortex of CUMS depressed mice

[0044] The level of IL-1β in the brain can be used to assess the level of neuroinflammation in the brain. Figure 4 As shown in a and b, compared with the CON group, the IL-1β levels in the hippocampus and cortex of the CUMS group mice were significantly increased (P < 0.01). Compared with the CUMS group, after IL and FLX treatment, the IL-1β levels in the hippocampus of CUMS mice were significantly reduced (IL 5mg / kg: P < 0.05; IL 10mg / kg: P < 0.01; FLX 10mg / kg: P < 0.05), and the IL-1β levels in the cortical tissue of mice were also significantly reduced (IL 5mg / kg: P < 0.01; IL 10mg / kg: P < 0.001; FLX 10mg / kg: P < 0.05). This indicates that isoliensinine alleviates the neuroinflammatory response in the hippocampus and cortex of CUMS depressed mice.

[0045] 5. Isoliensinine reduces serum CORT levels in CUMS depressed mice

[0046] Serum CORT levels can be used to preliminarily assess the hyperactivity of the HPA axis. Figure 5 As shown in the figure, compared with the CON group, the CORT level in the serum of the CUMS group mice was significantly increased (P < 0.01). Compared with the CUMS group, the CORT level in the serum of the CUMS mice was significantly reduced after IL and FLX treatment (IL 10 mg / kg: P < 0.001; FLX 10 mg / kg: P < 0.01). This indicates that isoliensinine can inhibit the hyperactivity of the HPA axis in CUMS depressed mice.

Claims

1. Application of isoliensinine in the preparation of antidepressant drugs.

2. The use according to claim 1, characterized in that: The structural formula of the isoliensinine is shown in (Ⅰ):

3. The use according to claim 1, characterized in that: The anti-depressant effects include improving depressive behaviors, alleviating inflammatory responses in the brain, and inhibiting hyperactivity of the HPA axis.

4. The use according to claim 1, characterized in that: The dosage form of the drug is any pharmaceutically therapeutically acceptable dosage form.

5. The use according to claim 1, characterized in that: The dosage of the drug is any therapeutically acceptable dosage.

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