Application of nomilin in preparation of medicine for treating and preventing depression

Abstract

The invention discloses application of nomilin in preparation of a medicine for treating and preventing depression, nomilin has a remarkable treatment effect on mouse depression-like behaviors induced by LPS (lipopolysaccharide), and nomilin can increase expression of PSD-95, SNAP-25 and Synaptophysin synaptic proteins so as to promote recovery of depression nerve synaptic structures and functions. The effects of resisting depression and resisting Alzheimer's disease are achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of functions and applications of natural small molecule compounds, and in particular to the application of nomilin in the preparation of drugs for treating and / or preventing depression. Background Art

[0002] Depression is characterized by a significant and persistent low mood, accompanied by a loss of interest and pleasure, which often affects an individual's work, study and social functions. It is one of the most serious diseases. Depression may be caused by a variety of factors, including genetic, biological, psychosocial and environmental factors. Although depression brings many harms, it is often not discovered in advance or treated accordingly, resulting in a significant decline in the quality of life of patients and their loved ones and causing great pain. This disease has caused significant harm to both individuals and society. Acute stress, as an important factor in the onset of depression, can lead to changes in neurodevelopment. Therefore, the study of the mechanism of the effect of acute stress on the onset of depression and the search for relevant drug interventions will provide strategies for the prevention and treatment of depressive disorders.

[0003] Synaptic loss and functional connectivity defects are considered to be important factors leading to depression. The synaptosome-associated protein SNAP-25 is one of the key proteins involved in the formation of the neurosoluble N-ethylmaleimide-sensitive factor attachment protein receptor complex and is also key to normal brain function. PSD-95 is a major regulator of synaptic maturation. PSD-95 is an important component involved in glutamatergic transmission, synaptic plasticity, and dendritic spine morphogenesis during neurodevelopment. PSD-95 dysfunction may change the synaptic plasticity of dendritic spines and lead to synaptic malformations associated with neurological diseases. Synaptophysin is a membrane protein closely related to synaptic function. In summary, PSD-95, SNAP-25, and Synaptophysin-related synaptic proteins are closely related to depression.

[0004] Nomilin is one of the main citric compounds and a secondary metabolite of triterpenoid plants. Nomilin is mainly found in common edible citrus fruits, including lemons, limes, oranges, grapefruits, and tangerines, as well as traditional Chinese medicines of citrus fruits, such as tangerines and tangerine peels. Citrus fruits are one of the largest crops produced worldwide, with grapefruit, lemons, oranges, and tangerines being among the most important crops.

[0005] Many studies have shown that nomilin and its analogs exhibit a variety of biological and pharmacological activities, including anti-cancer, immunomodulatory, anti-inflammatory, anti-obesity, anti-viral, anti-osteoclast, and anti-oxidation effects. Therefore, nomilin and its analogs have become potential treatments for human diseases. The present invention relates to the effects and efficacy of nomilin in antidepressant, and no relevant reports have been seen. Summary of the invention

[0006] The purpose of the present invention is to provide the use of Nomilin in preparing drugs for treating and / or preventing depression in view of the technical defects existing in the prior art.

[0007] Another object of the present invention is to provide the use of Nomilin in preparing drugs for treating and / or preventing Alzheimer's disease.

[0008] The technical solution adopted to achieve the purpose of the present invention is:

[0009] The use of Nomilin in the preparation of drugs for treating and / or preventing depression, the structural formula of Nomilin is as follows:

[0010]

[0011] In the above technical solution, the depression is LPS-induced acute stress depression.

[0012] In the above technical scheme, Nomilin inhibits LPS-induced acute stress depression by increasing the expression of synapse-related proteins.

[0013] Another aspect of the present invention also includes the use of Nomilin in the preparation of drugs for treating and / or preventing Alzheimer's disease. The structural formula of Nomilin is as follows:

[0014]

[0015] In the above technical solution, Nomilin inhibits the Alzheimer's disease by increasing the expression of synapse-related proteins.

[0016] Another aspect of the present invention also includes a drug for treating and / or preventing depression, wherein the active ingredient includes Nomilin, and the structural formula of Nomilin is as follows:

[0017]

[0018] Another aspect of the present invention also includes a drug for treating and / or preventing Alzheimer's disease, wherein the active ingredient includes Nomilin, and the structural formula of Nomilin is as follows:

[0019]

[0020] In the above technical solution, the medicine also contains pharmaceutically acceptable excipients.

[0021] In the above technical solution, the auxiliary materials include diluents, excipients, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers or lubricants.

[0022] In the above technical solution, the dosage form of the drug is capsule, microcapsule, pill, tablet, decoction, granule, paste, dispersible powder, oral dew, pill or liposome.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] 1. Nomilin alleviates LPS-induced depressive-like behavior in mice: Intraperitoneal injection of LPS into mice can produce acute stress-induced depressive-like behavior. The antidepressant effect of Nomilin was explored in this model by performing SPT, TST and FST tests. Compared with mice in the control group, mice treated with LPS showed less sucrose consumption, while pretreatment with Nomilin significantly increased the sucrose consumption of mice. Similar results were also observed in mice pretreated with Fluoxetine. In TST, mice treated with LPS caused a significant increase in immobility time compared with mice in the control group. In contrast, pretreatment with Nomilin and pretreatment with Fluoxetine significantly reduced immobility time compared with mice treated with LPS. Similar results were observed in FST, and the increase in immobility time of mice treated with LPS was significantly inhibited by pretreatment with Nomilin and Fluoxetine. These results indicate that Nomilin has a significant therapeutic effect on LPS-induced depressive-like behavior in mice.

[0025] 2. Nomilin increases the expression of PSD-95, SNAP-25, and Synaptophysin related synaptic proteins: Western blot analysis of LPS-induced mouse brain tissue showed that the expression of PSD-95, SNAP-25, and Synaptophysin proteins in the LPS stimulation group was significantly reduced compared with the control group, while the results of the Nomilin treatment group were reversed. This indicates that Nomilin can increase the expression of PSD-95, SNAP-25, and Synaptophysin synaptic proteins, thereby promoting the recovery of the structure and function of synapses in depression, and play an anti-depression and anti-Alzheimer's role. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Effects of Nomilin on sugar preference in depression model mice ## P<0.01 vs control group; **P<0.01 vs LPS model group).

[0027] Figure 2 The results of the tail suspension experiment of Nomilin on depression model mice ( ## P<0.01 vs control group; *P<0.05, **P<0.01 vs LPS model group).

[0028] Figure 3 The results of the forced swimming test of normilin on depression model mice (## P<0.01 vs control group; **P<0.01 vs LPS model group).

[0029] Figure 4 The effect of Nomilin on the expression of PSD-95 in the cortical tissue of depression model mice and its quantitative diagram ( # P<0.05 vs control group; **P<0.01 vs LPS model group).

[0030] Figure 5 The effect of Nomilin on the expression of SNAP-25 in the cortical tissue of depression model mice and its quantitative diagram ( # P<0.05 vs control group; **P<0.01 vs LPS model group).

[0031] Figure 6 The effect of Nomilin on the expression of Synaptophysin in the cortical tissue of depression model mice and its quantification ( ## P<0.01 vs control group; **P<0.01 vs LPS model group). DETAILED DESCRIPTION

[0032] The present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0033] Establishment of acute depression model in mice:

[0034] Male C57BL / 6 mice were housed in a temperature- and relative humidity-controlled facility with a 12-h light-dark cycle and provided with adequate food and water. After acclimation, they were randomly divided into 4 groups: control group, LPS model group, LPS+nomilin (50 mg / kg) group, and LPS+fluoxetine group (20 mg / kg). In the LPS+nomilin (50 mg / kg) group, nomilin (dissolved in 0.5% sodium carboxymethylcellulose) was administered by gavage, and in the LPS+fluoxetine group (20 mg / kg), fluoxetine (dissolved in normal saline) was intraperitoneally injected once a day for 4 consecutive days. The control group and LPS model group were given the same amount of solvent (0.5% sodium carboxymethylcellulose) by gavage. Starting from the second day, the LPS model group, LPS+nomine group and LPS+fluoxetine group were intraperitoneally injected with LPS (1 mg / kg) once a day for 3 consecutive days to establish a mouse depression model. The control group was intraperitoneally injected with the same volume of normal saline. After the last injection of LPS, behavioral tests were performed. After the behavioral tests were completed, the mouse cortical tissues were taken for subsequent tests.

[0035] Sugar Preference Test (SPT):

[0036] The mice were adapted to sugar and water respectively. After adaptation, they were deprived of food and water for 12 hours. Then two water bottles were placed, one filled with water and the other with 1% sucrose solution. The weight of sucrose solution and water within 12 hours after the last LPS injection was recorded, and the consumption was calculated using sucrose preference (%) = sucrose consumption / (sucrose consumption + water consumption) × 100%.

[0037] Tail suspension test (TST):

[0038] The mice were individually suspended 50 cm above a flat surface and taped approximately 1 cm from the tip of the tail. Each mouse was suspended for 6 min, and the immobility time of each mouse was recorded for 4 min.

[0039] Forced swim test (FST):

[0040] The mice were placed individually in a transparent cylinder (a cylindrical transparent glass container with a diameter of 15 cm and a height of 30 cm) filled with fresh water. The water temperature was maintained at 25±1°C. The water droplets on the inner and outer walls of the container were wiped clean. Each mouse was forced to swim for 6 minutes, and the immobility time of each mouse was recorded for 4 minutes.

[0041] Western blot analysis:

[0042] According to the weight of the mouse, add the corresponding volume of RIPA lysis buffer, grind into a homogenate, collect the lysate, and then centrifuge at 12000rpm for 20 minutes. Collect the supernatant and determine the protein concentration using the BCA protein detection kit. Then add the loading buffer to each sample by volume, heat for 10 minutes, separate the protein using 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and transfer the protein to a PVDF membrane. The membrane was incubated with 3% BSA at room temperature for 2h, and then incubated with the primary antibody at 4℃ overnight. The next day, wash the membrane 3 times with TBST, incubate with the secondary antibody for 2h, and wash 3 times with TBST. Finally, the target protein bands were detected using an ECL imaging system.

[0043] like Figure 1-Figure 3 , Nomilin can significantly reduce the depressive-like behavior of acutely stressed mice induced by LPS. Specifically, it was shown by SPT, TST and FST tests. In SPT, compared with mice in the control group, mice in the LPS model group showed less sucrose consumption, while pretreatment with Nomilin significantly increased the sucrose consumption of mice. Similar results were also observed in mice pretreated with fluoxetine. In TST, compared with mice in the control group, mice in the LPS model group showed a significantly increased immobility time. In contrast, compared with mice in the LPS model group, the Nomilin and fluoxetine treatment groups significantly reduced the immobility time of mice. Similar results were observed in FST. These results indicate that Nomilin has a significant therapeutic effect on the depressive-like behavior of mice induced by LPS.

[0044] like Figure 4-Figure 6 , protein detection showed that Nomilin can significantly increase the expression of PSD-95, SNAP-25, and Synaptophysin related synaptic proteins in LPS-induced mouse brain tissue. Specifically, we performed protein blotting analysis on LPS-induced mouse brain tissue. Compared with the control group, the expression of PSD-95, SNAP-25, and Synaptophysin proteins in the LPS model group was significantly reduced, while the results of the LPS+Nomilin (50mg / kg) group were reversed. This shows that Nomilin can increase the expression of PSD-95, SNAP-25, and Synaptophysin related synaptic proteins, thereby promoting the recovery of the structure and function of depressive synapses, and play an anti-depressant and anti-Alzheimer's role.

[0045] The above is only a preferred embodiment of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. The use of Nomilin in preparing a drug for treating and / or preventing depression, characterized in that: The structural formula of Nomilin is as follows:

2. The use according to claim 1, characterized in that The depression is LPS-induced acute stress depression.

3. The use according to claim 2, characterized in that Nomilin inhibits LPS-induced acute stress depression by increasing the expression of synapse-related proteins.

4. The use of Nomilin in the preparation of a drug for treating and / or preventing Alzheimer's disease, characterized in that: The structural formula of Nomilin is as follows:

5. The use according to claim 4, characterized in that Nomilin inhibits the Alzheimer's disease by increasing the expression of synapse-related proteins.

6. A drug for treating and / or preventing Alzheimer's disease, characterized in that: The active ingredient includes Nomilin, and the structural formula of Nomilin is as follows:

7. A drug for treating and / or preventing depression, characterized in that: The active ingredient includes Nomilin, and the structural formula of Nomilin is as follows:

8. The drug according to claim 6 or 7, characterized in that The medicine also contains pharmaceutically acceptable excipients.

9. The drug according to claim 6 or 7, characterized in that The auxiliary materials include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers or lubricants.

10. The drug according to claim 6 or 7, characterized in that The dosage form of the drug is capsule, microcapsule, pill, tablet, decoction, granule, paste, dispersible powder, oral dew, drop pill or liposome.

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