Application of calculus bovis in preparation of anti-depression product

Abstract

The invention provides application of calculus bovis in preparation of an anti-depression product, and belongs to the technical field of medicines. The invention provides application of calculus bovis in preparation of an anti-depression product. Reserpine and CUMS depression mouse models are used, it is verified that ICCB can remarkably improve depression-like characterization of two animal models of reserpine and CUMS, it is proved for the first time that ICCB has the anti-depression effect, meanwhile, pathological damage of hippocampal neurons of mice after modeling can be effectively inhibited, various indexes of the liver function of the model mice can be remarkably recovered, and the ICCB has the anti-depression effect. The ICCB has the effect of regulating the blood fat level remarkably related to liver metabolism, and particularly has the effect of regulating the blood fat level remarkably related to liver metabolism, so that the ICCB not only improves depression-like characterization and pathological damage of two animal models of reserpine and CUMS, but also has a certain protection effect on liver functions and blood fat metabolism functions which are reduced after modeling.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to the application of bezoar in preparing antidepressant products. Background Art

[0002] Depression is a mental illness characterized by long-term low mood, slow thinking, weakened willpower and cognitive dysfunction. It is difficult to cure and prone to relapse. In severe cases, it may even lead to suicidal tendencies. However, existing treatment drugs take a long time to take effect and also produce many adverse reactions when taken. Therefore, it is of great practical significance to develop depression treatment drugs with clear efficacy and few side effects.

[0003] Bezoar is the dried gallstone of cattle (Bos taurus domesticus Gmelin), a traditional precious medicinal material in my country. It has many pharmacological effects such as expectoration, opening the mind, and detoxification. However, due to the shortage of drug sources, it has been difficult to meet the needs of clinical medication for a long time. In vitro cultured calculus bovis (ICCB) is included in the 2020 edition of the Chinese Pharmacopoeia. It simulates the principle and biochemical process of gallstone formation in vivo. It uses bioengineering technology to cultivate bovine bilirubin calcium stones in bovine bile in vitro. Its properties, composition, content, structure, and clinical efficacy are similar to those of high-quality natural bezoar. It has been widely used in production as a substitute for natural bezoar. Modern pharmacological studies have shown that ICCB has multiple pharmacological effects such as antioxidant, antibacterial and anti-inflammatory, and immune function enhancement, but whether it has an antidepressant effect has not been reported. Summary of the invention

[0004] In view of the defects in the prior art, the purpose of the present invention is to provide a use of bezoar in the preparation of an antidepressant product, wherein the bezoar can be used to treat depression with significant therapeutic effect.

[0005] The objective of the present invention is achieved through the following technical solutions:

[0006] The invention provides application of bezoar in preparing antidepressant products.

[0007] Preferably, the application includes any one or more of the following (1) to (6):

[0008] (1) Improve weight changes;

[0009] (2) Improve body temperature changes;

[0010] (3) Improve behavioral changes;

[0011] (4) inhibiting hippocampal neuron damage;

[0012] (5) Improve liver function;

[0013] (6) Improve lipid metabolism function.

[0014] Preferably, the inhibition of hippocampal neuron damage includes: reducing the number of damaged cells and / or preventing neuronal synapse damage.

[0015] Preferably, the improvement of liver function includes: any one or two or more of inhibiting the increase of AST, inhibiting the increase of ALT and inhibiting the decrease of ALP.

[0016] Preferably, the improvement of lipid metabolism function includes: significantly increasing the content of any one or more of CHOL, HDL-C, LDL-C and TG.

[0017] Preferably, the depression comprises drug-induced depression and / or depression induced by changes in environmental conditions.

[0018] Preferably, the bezoar comprises bezoar cultivated in vitro.

[0019] Preferably, the product comprises a pharmaceutical product.

[0020] Preferably, the medicine further comprises excipients.

[0021] Preferably, the excipients include pharmaceutically acceptable excipients.

[0022] Beneficial effects of the present invention:

[0023] The present invention provides the use of bezoar in the preparation of antidepressant products. The present invention uses reserpine and CUMS depression mouse models to verify that ICCB can significantly improve the depressive-like manifestations of the two animal models of reserpine and CUMS, and proves for the first time that ICCB has the effect of antidepressant, and can effectively inhibit the pathological damage of hippocampal neurons in mice after modeling, and significantly restore various indicators of liver function in model mice, especially the regulatory effect of ICCB on blood lipid levels significantly related to liver metabolism, indicating that ICCB not only improves the depressive-like manifestations and pathological damage of the two animal models of reserpine and CUMS, but also has a certain protective effect on the liver function and blood lipid metabolism function that are downregulated after modeling. In summary, the bezoar can be used to treat depression, and the therapeutic effect is significant. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0025] Figure 1This is the effect of ICCB on body weight in the reserpine depression mouse model;

[0026] Figure 2 This is a graph showing the effect of ICCB on body temperature in the reserpine depression mouse model;

[0027] Figure 3 This is the effect of ICCB on the sugar preference index of the reserpine depression mouse model;

[0028] Figure 4 This is the effect of ICCB on the tail suspension immobility time in the reserpine depression mouse model;

[0029] Figure 5 This is a graph showing the effect of ICCB on the swimming immobility time in the reserpine depression mouse model;

[0030] Figure 6 The figure shows the effect of ICCB on the damage of CA1, CA3 and DG areas of the hippocampus in the reserpine depression mouse model;

[0031] Figure 7 This is the observation diagram of ICCB on the synaptic damage of neurons in the CA1 region of the mouse hippocampus induced by reserpine;

[0032] Figure 8 This is the statistical result of ICCB on the number of dendritic spines in the CA1 region of the mouse hippocampus induced by reserpine;

[0033] Fig. 9 This is the effect of ICCB on AST in reserpine model mice;

[0034] Fig.10 The figure shows the effect of ICCB on ALT in reserpine model mice;

[0035] Fig.11 The figure shows the effect of ICCB on ALP in reserpine model mice;

[0036] Fig.12 The figure shows the effect of ICCB on CHOL in reserpine model mice;

[0037] Fig.13 The figure shows the effect of ICCB on HDL-C in reserpine model mice;

[0038] Fig.14 The figure shows the effect of ICCB on LDL-C in reserpine model mice;

[0039] Fig.15 The figure shows the effect of ICCB on TG in reserpine model mice;

[0040] Fig.16 This is the effect of ICCB on the sugar preference index of the CUMS depression mouse model;

[0041] Fig.17 This is the effect of ICCB on the tail suspension immobility time in the CUMS depression mouse model;

[0042] Fig.18 This is the effect of ICCB on the swimming immobility time in the CUMS depression mouse model;

[0043] Fig.19 The effect of ICCB on the damage of CA1, CA3 and DG areas of the hippocampus in the CUMS depression mouse model;

[0044] Fig. 20 This is a diagram showing the effect of ICCB on AST in CUMS model mice;

[0045] Fig.21 The figure shows the effect of ICCB on ALT in CUMS model mice;

[0046] Fig. 22 The figure shows the effect of ICCB on ALP in CUMS model mice;

[0047] Fig.23 The figure shows the effect of ICCB on CHOL in CUMS model mice;

[0048] Fig.24 This is the effect of ICCB on HDL-C in CUMS model mice;

[0049] Fig.25 The figure shows the effect of ICCB on LDL-C in CUMS model mice;

[0050] Fig.26 This is a diagram showing the effect of ICCB on TG in CUMS model mice. DETAILED DESCRIPTION

[0051] The invention provides application of bezoar in preparing antidepressant products.

[0052] In the present invention, the anti-depression includes preventing and / or treating depression.

[0053] The present invention has no special limitation on the source of the bezoar, and any conventional source in the art can be used. As an optional embodiment of the present invention, the bezoar can be in vitro cultured bezoar.

[0054] In the present invention, the product includes a medicine; the medicine may further include an excipient; the excipient includes a pharmaceutically acceptable excipient.

[0055] In the present invention, the depression includes drug-induced depression and / or depression induced by changing environmental conditions. In the embodiments of the present invention, the depression model induced by reserpine or the CUMS depression mouse model is taken as an example to verify the therapeutic effect of bezoar on depression. The present invention uses reserpine and CUMS depression mouse models to verify that ICCB can significantly improve the depressive-like manifestations of the two animal models of reserpine and CUMS. It is the first time to prove that ICCB has the effect of anti-depression, and can effectively inhibit the pathological damage of the hippocampus of mice after modeling, and significantly restore the various indicators of liver function of model mice, especially the regulatory effect of ICCB on blood lipid levels that are significantly related to liver metabolism, indicating that ICCB not only improves the depressive-like manifestations and pathological damage of the two animal models of reserpine and CUMS, but also has a certain protective effect on the liver function and blood lipid metabolism function that are downregulated after modeling.

[0056] In the present invention, the medicine includes a unit dose preparation, and the medicine contains an effective dose of bezoar. The medicine is formulated into a dosage form for administration of 10 to 60 mg / kg / d based on the body weight of the mouse model. In an embodiment of the present invention, the dosage of bezoar can be 10 to 60 mg / kg / d, or 35 to 60 mg / kg / d; as an optional embodiment of the present invention, the dosage of bezoar can be 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 mg / kg / d.

[0057] In the present invention, the application includes any one or more of the following (1) to (6):

[0058] (1) Improve weight changes;

[0059] (2) Improve body temperature changes;

[0060] (3) Improve behavioral changes;

[0061] (4) inhibiting hippocampal neuron damage;

[0062] (5) Improve liver function;

[0063] (6) Improve lipid metabolism function.

[0064] In the present invention, the bezoar can improve the weight change caused by depression. The present invention shows through the results of the examples that the in vitro cultured bezoar can significantly improve the weight change of the reserpine-induced depression mouse model, making its weight change tend to normal level.

[0065] In the present invention, the bezoar can improve the temperature change caused by depression. The present invention shows through the example results that the in vitro cultured bezoar can significantly improve the temperature change of the reserpine-induced depression mouse model, making its temperature change tend to normal level.

[0066] In the present invention, the bezoar can improve behavioral changes caused by depression. The present invention shows through the results of the examples that the in vitro cultured bezoar can significantly improve the sugar preference index, tail suspension immobility time and swimming immobility time of the reserpine and CUMS depression mouse models, and effectively improve the depressive-like behavior of the reserpine model mice.

[0067] In the present invention, the bezoar can inhibit hippocampal neuron damage. In the present invention, the inhibition of hippocampal neuron damage includes: reducing the number of damaged cells and / or preventing neuronal synaptic damage. The present invention shows through the results of the examples that the in vitro cultured bezoar can significantly reduce the number of damaged cells in the CA1, CA3 and DG regions of the hippocampal neurons in the reserpine and CUMS depression mouse models, and can significantly prevent the reserpine-induced neuronal synaptic damage in the CA1 region of the hippocampus.

[0068] In the present invention, the bezoar can improve liver function. In the present invention, the improvement of liver function includes: inhibiting the increase of AST, inhibiting the increase of ALT and inhibiting the decrease of ALP or more. The present invention shows through the results of the examples that the in vitro cultured bezoar can significantly downregulate the increase of AST and ALT in the reserpine and CUMS depression mouse models, while significantly recovering the ALP content.

[0069] In the present invention, the bezoar can improve lipid metabolism function. The present invention shows through the example results that the in vitro cultured bezoar can significantly increase the CHOL, HDL-C, LDL-C and TG contents of the reserpine and CUMS depression mouse models.

[0070] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0071] The method of data statistical analysis in the following embodiments:

[0072] Statistical analysis was performed using SPSS 19.0, and statistical results were plotted using GraphPad Prism 8.0, and presented as mean ± standard deviation (±s). One-way ANOVA was used to analyze the differences between groups. P value represents significance, with P < 0.05 considered to be significantly different, P < 0.01 considered to be relatively significantly different, and P < 0.001 considered to be extremely significantly different.

[0073] Example 1

[0074] The therapeutic effect of in vitro cultured bezoar on drug-induced depression model

[0075] 1. Construction of reserpine depression mouse model

[0076] Sixty healthy ICR male mice were selected and, after one week of adaptive feeding, were randomly divided into six groups: normal group: intragastric administration of normal saline on days 0-7; model group: intragastric administration of normal saline on days 0-7, and intraperitoneal injection of 1.25 mg / kg / d reserpine on days 5-7; imipramine group: intragastric administration of 30 mg / kg / d imipramine on days 0-7, and intraperitoneal injection of 1.25 mg / kg / d reserpine on days 5-7; low-dose ICCB group: intragastric administration of 10 mg / kg / d ICCB on days 0-7, and intraperitoneal injection of 1.25 mg / kg / d reserpine on days 5-7; medium-dose ICCB group: intragastric administration of 35 mg / kg / d ICCB on days 0-7, and intraperitoneal injection of 1.25 mg / kg / d reserpine on days 5-7; high-dose ICCB group: intragastric administration of 60 mg / kg / d on days 0-7 ICCB, intraperitoneal injection of 1.25 mg / kg / d reserpine on days 5-7.

[0077] ICCB was purchased from Wuhan Jianmin Dapeng Pharmaceutical Co., Ltd., batch number: 22120, and the solvent for dissolving ICCB was 0.5% sodium carboxymethylcellulose saline solution. The volume of normal saline gavage in the normal group, the volume of saline gavage in the model group, the volume of imipramine solution gavage, and the volume of ICCB solution gavage were all converted according to the volume of 20g / 0.3mL. Among them, 20g / 0.3mL means that the volume of the gavage solution for mice weighing 20g is 0.3mL, the same below.

[0078] 2. Body weight and temperature detection of reserpine depression mouse model after ICCB administration

[0079] From the 5th day to the 8th day, the weight and temperature of the mice were tested every day. The weight and temperature of the mice were tested and recorded before each reserpine injection. The weight and temperature of the mice were measured at the same time on the 8th day, and the differences in weight and temperature of the mice in each group were counted every day. The weight was measured using an electronic balance; the temperature was measured using a soft-headed electronic thermometer inserted 1 cm from the periphery of the mouse anus. The thermometer was lubricated with paraffin before insertion, and the temperature was recorded after the reading stabilized.

[0080] The results of ICCB reducing the effects of reserpine on the body temperature and body weight of mice are shown in Tables 1-2 and Figures 1-2 As shown. Among them, Figure 1 The effect of ICCB on body weight in the reserpine depression mouse model; Figure 2 The effect of ICCB on body temperature in the reserpine depression mouse model.

[0081] Table 1 Effect of ICCB on body weight of reserpine depression mouse model ( n=10)

[0082] Group Day 5 (g) Day 6 (g) Day 7 (g) Day 8 (g) normal 29.53±1.10 29.93±1.24 30.19±1.42 30.70±1.52 Model 28.98±1.89 <![CDATA[28.02±1.91 # ]]> <![CDATA[26.83±2.02 ### ]]> <![CDATA[24.13±2.07 ### ]]> Imipramine 30 mg / kg 29.76±1.97 29.06±1.79 28.52±2.08 <![CDATA[26.63±2.25 * ]]> ICCB10mg / kg 29.70±1.62 28.97±1.58 27.92±1.90 25.36±2.38 ICCB35mg / kg 29.75±1.85 29.44±1.84 28.69±2.11 <![CDATA[26.59±2.26 $ ]]> ICCB 60mg / kg 29.73±1.70 29.03±1.71 28.33±2.41 26.04±2.99

[0083] Note: Compared with the model group, ICCB 35mg / kg $ P<0.05; compared with the model group, * P<0.05; compared with the model group and the normal group, # P<0.05, ### P<0.001.

[0084] Table 2 Effects of ICCB on body temperature in the reserpine depression mouse model ( n=10)

[0085] Group Day 5 (g) Day 6 (g) Day 7 (g) Day 8 (g) normal 37.05±0.27 37.08±0.23 37.33±0.16 37.22±0.21 Model 37.04±0.24 <![CDATA[36.13±0.57 ## ]]> <![CDATA[35.67±0.64 ### ]]> <![CDATA[35.04±1.00 ### ]]> Imipramine 30 mg / kg 37.00±0.16 36.56±0.15 36.30±0.10 <![CDATA[36.00±0.19 ** ]]> ICCB10mg / kg 37.06±0.26 36.31±0.20 36.10±0.37 35.52±0.26 ICCB35mg / kg 37.12±0.23 36.52±0.23 36.27±0.21 <![CDATA[36.02±0.28 $ ]]> ICCB 60mg / kg 37.12±0.16 36.48±0.37 36.20±0.17 <![CDATA[35.88±0.08 & ]]>

[0086] Note: Compared with the model group, ICCB 35mg / kg $ P<0.05; ICCB 60mg / kg compared with the model group, & P<0.05; compared with the model group, ** P<0.01; compared with the model group and the normal group, ## P<0.01, ### P<0.001.

[0087] From Tables 1 to 2 and Figures 1-2 It was found that from the first day after reserpine injection, the body weight and body temperature of mice in the model group decreased significantly compared with those in the normal group (P<0.05, P<0.01), and showed a very significant downward trend on the second and third days (P<0.001), and the mice showed obvious emaciation. Compared with the model group, the weight loss trend of the imipramine group and the medium-dose ICCB group on the third day was significantly weakened (P<0.05), and the downward trend of the body temperature of mice in the high-dose and medium-dose ICCB groups was significantly weakened (P<0.05), and the downward trend of the body temperature of mice in the imipramine group was more significantly weakened (P<0.01).

[0088] 3. Observation of behavioral changes in the reserpine depression mouse model after ICCB intervention

[0089] The behavioral changes of the following groups of mice were detected within 3 days after the reserpine model was established:

[0090] (1) Sugar water consumption experiment: Each mouse was given two bottles of water with a predetermined amount of water: one bottle of 1% sucrose water and one bottle of pure water. The mice were allowed to adapt for 48 hours, and the positions of the water bottles were swapped every 12 hours. After the mice were deprived of water for 12 hours, the food was removed and the mice were given one bottle of 1% sucrose water and one bottle of pure water with a predetermined amount of water. The positions of the water bottles were swapped at 1 hour, 2 hours, and 6 hours. The total liquid consumption and sugar water consumption ratio of the mice within 12 hours were calculated.

[0091] Sugar water preference index (%) = [sugar water consumption / (sugar water consumption + water consumption)] × 100%

[0092] (2) Tail suspension test: The mouse’s tail was fixed to a high rack with tape at 1 / 3 of its length, with the head about 15 cm above the table. The mouse’s activity was recorded for 5 minutes.

[0093] (3) Forced swimming test: Using the FST forced swimming apparatus, mice were placed in a glass tank with a diameter of 18 cm and filled with 25°C tap water. The software recorded the immobility time within 5 min.

[0094] The effect of ICCB on improving the depression-like phenotype of mice induced by reserpine is shown in Tables 3 and Figures 3 to 5 shown. Figure 3 The effect of ICCB on the sugar preference index of the reserpine depression mouse model; Figure 4 The effect of ICCB on the tail suspension immobility time in the reserpine depression mouse model; Figure 5 The effect of ICCB on the swimming immobility time in the reserpine depression mouse model.

[0095] Table 3 Effects of ICCB on sugar preference index, tail suspension immobility time and swimming immobility time in the reserpine depression mouse model ( n=10)

[0096] Group Sugar water preference index (%) Tail suspension immobility time (s) Swimming immobility time (s) normal 81.11±6.48 48.40±35.56 167.57±20.1607 Model <![CDATA[50.35±10.74 ### ]]> <![CDATA[230.54±37.44 ### ]]> <![CDATA[222.11±19.29 ### ]]> Imipramine 30 mg / kg <![CDATA[66.19±9.49 ** ]]> <![CDATA[187.58±35.48 ** ]]> <![CDATA[179.61±18.82 *** ]]> ICCB10mg / kg 56.20±8.30 219.46±35.72 209.79±20.81 ICCB35mg / kg <![CDATA[65.36±7.38 ** ]]> <![CDATA[186.67±33.24 ** ]]> <![CDATA[178.59±29.11 *** ]]> ICCB 60mg / kg <![CDATA[59.09±5.78 * ]]> <![CDATA[188.46±33.00 * ]]> <![CDATA[191.36±22.06 ** ]]>

[0097] Note: Compared with the model group, * P<0.05, ** P<0.01, *** P<0.001; compared with the normal group, ### P<0.001.

[0098] From Table 3 and Figures 3 to 5It was found that compared with the normal group mice, the sugar preference index, tail suspension immobility time and swimming immobility time of the model group mice all changed significantly (P<0.001). Compared with the model group, the sugar preference index of the mice in the imipramine group and the medium-dose group was significantly increased (P<0.01), and the high-dose group was significantly increased (P<0.05); the tail suspension and swimming immobility time of the mice in the imipramine group and the medium-dose group were significantly decreased (P<0.01, P<0.001), while the high-dose group was significantly decreased (P<0.05, P<0.01). The results showed that ICCB can effectively improve the depressive-like behavior of reserpine model mice. When the drug concentration is too high, due to the complex composition of ICCB itself, it cannot be ruled out that the increase in the concentration of other components may affect the therapeutic effect at its appropriate concentration, so the therapeutic effect of high-dose ICCB may be slightly weaker than that of medium dose.

[0099] 4. HE staining to observe the effect of ICCB on hippocampal neuron damage in the reserpine depression mouse model

[0100] (1) After the behavioral test, mice in the normal group, model group, imipramine group, and medium-dose ICCB group were anesthetized with Shutai 50 (75 mg / kg, intramuscular injection) and fixed. They were placed in a supine position with their abdomen facing upwards, and their abdominal cavity was quickly exposed. The abdominal aorta was clamped with hemostats, and the chest cavity was opened to fully expose the heart. The apex of the heart was cut, and the perfusion needle was quickly inserted through the left ventricle to the ascending aorta. The right atrial appendage was cut and perfused quickly with normal saline to flush the blood in the mouse body. The perfusion was performed until the liver turned white and the liquid flowing out of the right atrial appendage was clear. Freshly prepared 4% paraformaldehyde was used for slow perfusion until the mouse's limbs turned pale and the organs of the whole body became hard. The brain tissue was removed and fixed in 4% paraformaldehyde at 4°C for 24 hours.

[0101] (2) Remove the brain tissue from the 4% paraformaldehyde and rinse it with running water for 2 hours to remove the residual paraformaldehyde. Then dehydrate it with ethanol step by step, in the order of 50% ethanol (2 hours), 60% ethanol (2 hours), 75% ethanol (overnight), 85% ethanol (3 hours), 95% ethanol (I) (1 hour), 95% ethanol (II) (1 hour), anhydrous ethanol (I) (1 hour), anhydrous ethanol (II) (1 hour). The dehydration time of 95% ethanol and anhydrous ethanol should not be too long, so as to avoid excessive dehydration and brittleness of the tissue. After dehydration, perform gradient transparency, in the order of 1 / 2 anhydrous ethanol + 1 / 2 xylene (10 minutes), pure xylene (I) (15 minutes), and pure xylene (II) (15 minutes). When transparentizing, ensure that the tissue is completely transparent to light before proceeding to the next step.

[0102] (3) Wax immersion and embedding: The wax immersion temperature and concentration requirements are from low to high. The tissue is taken out of pure xylene and placed in a wax immersion system formed by 1 / 2 xylene + 1 / 2 paraffin for 1 hour. The temperature of the wax immersion system is maintained at 54-56°C; then placed in pure paraffin at 54-56°C for 2 hours; and then placed in pure paraffin at 56-58°C for 2 hours. To prevent the paraffin from solidifying or incomplete wax immersion, the wax immersion process should maintain a constant temperature and be 2-3°C higher than the melting point. Set the temperature of the wax cylinder, wax nozzle, wax stand, left and right cylinders of the embedding machine to 60°C, 65°C, 60°C, and 60°C. Place the wax-immersed tissue in a metal mold, cover the surface of the tissue with the paraffin flowing out of the wax nozzle, and after cooling slightly, cover the plastic mold and place it on a freezing table to cool.

[0103] (4) Slicing, spreading, sticking and drying: Fix the embedded paraffin block on the slicer and trim it. The slice thickness should be 6 μm. Place the cut paraffin slices in 40℃ warm water and spread them. Pick up the slices at 45° so that the paraffin slices can be completely spread on the sticking slide. Place the sticking tissue slide on the slide rack and dry it at room temperature so that the tissue is completely stuck to the slide.

[0104] (5) HE staining: After the paraffin sections of brain tissue were prepared, they were stained with an automatic stainer, sealed with neutral gum, and the status of the neurons in each subregion of the hippocampus was observed using a NanoZoomer digital slice scanner.

[0105] Effects of the optimal dose of 35 mg / kg ICCB on the damage of the hippocampal CA1, CA3 and DG regions in the reserpine depression mouse model Figure 6 shown. Figure 6 Middle scale bar = 100 μm.

[0106] like Figure 6 As shown in the figure, the neurons in the CA1, CA3 and DG regions of the hippocampus of the mice in the normal group had regular morphology, plump cell bodies and close arrangement. There were more damaged cells in the CA1, CA3 and DG regions of the hippocampus of the mice in the model group, which were characterized by dark staining of the nucleus, shrinkage and deformation of the cells, triangular or irregular shapes, and blurred boundaries between the nucleus and cytoplasm. Compared with the model group, the number of damaged cells in each division of the hippocampus of the mice in the imipramine group and the medium-dose group was significantly reduced, indicating that ICCB effectively inhibited the damage of hippocampal neurons in the reserpine mouse model.

[0107] 5. Golgi staining to observe the changes in the number of dendritic spines of hippocampal neurons in the reserpine depression mouse model after ICCB administration

[0108] Three mice from each of the normal group, model group, imipramine group, and medium-dose ICCB group were anesthetized and the intact brain tissue was removed. The blood on the surface was quickly washed off with distilled water, and the mouse brain tissue was directly placed in the Golgi staining fixative in a fume hood. After fixation for 3 days, the brain tissue was transferred to the staining solution and placed in a cool and ventilated place at room temperature and away from light for 14 days (a new staining solution was changed after immersion for 48 hours, and then a new staining solution was changed every 3 days). On the 14th day, the tissue treatment solution was replaced and immersed in the dark at 4°C for 3 days.

[0109] The processed brain tissue was glued to the tray of the vibrating slicer with a slice thickness of 60 μm. After the slices were sliced, they were attached to the glass slide with the help of a brush. Tissue treatment solution was dripped on the surface of the brain tissue slices. After drying slightly, Golgi developer was dripped. After 30 minutes of development, the slices were washed with water and sealed, and pictures were taken with a tissue slice digital scanner. After obtaining the pictures, a 50 μm complete neuronal synapse was randomly selected from the CA1 region of the left and right hippocampus of each group of mice, and the pictures were imported into Image J and the number of dendritic spines was recorded. The protective effect of ICCB on reserpine-induced synaptic damage of neurons in the CA1 region of the mouse hippocampus is shown in Tables 4 and Figures 7-8 shown. Figure 7 This is the observation diagram of ICCB on the synaptic damage of neurons in the CA1 region of the mouse hippocampus induced by reserpine. Figure 7 The scale bar in the figure is 5 μm. Figure 8 This is the statistical result of the effect of ICCB on the number of dendritic spines in the CA1 region of the mouse hippocampus induced by reserpine.

[0110] Table 4 Protective effect of ICCB on synaptic damage of neurons in the CA1 region of the hippocampus of mice induced by reserpine ( n=3)

[0111]

[0112]

[0113] Note: Compared with the model group, * P<0.05; compared with the normal group, ## P<0.01.

[0114] From Table 4 and Figures 7-8 It was found that compared with the normal group mice, the number of dendritic spines in the CA1 region of the hippocampus of the model group mice was significantly reduced (P<0.01); the advance administration of imipramine and 35 mg / kg ICCB could significantly prevent the synaptic damage of the CA1 region of the hippocampus neurons induced by reserpine (P<0.05).

[0115] 6. Effects of ICCB on liver function and blood lipids in the reserpine depression mouse model detected by blood analyzer

[0116] After another batch of mice were divided into groups and modeling was completed, blood was immediately collected from the eyeballs of mice in the normal group, model group, imipramine group, and medium-dose ICCB group. The plasma was placed at room temperature for 1 hour and then centrifuged at 1000 rpm to obtain the supernatant (>200 μL / sample) and sent to the Animal Experiment Center of Zhejiang Chinese Medical University for determination of AST, ALT, ALP, CHOL, HDL-C, LDL-C, and TG using an automatic blood analyzer.

[0117] The effects of ICCB on the changes of liver function indexes and blood lipid parameters in reserpine model mice are shown in Table 5 and Figures 9 to 15 As shown. Fig. 9 The effect of ICCB on AST in reserpine model mice; Fig.10 The effect of ICCB on ALT in reserpine model mice; Fig.11 The effect of ICCB on ALP in reserpine model mice; Fig.12 The effect of ICCB on CHOL in reserpine model mice; Fig.13 The effect of ICCB on HDL-C in reserpine model mice; Fig.14 The effect of ICCB on LDL-C in reserpine model mice; Fig.15 The effect of ICCB on TG in reserpine model mice.

[0118] Table 5 Effects of ICCB on changes in liver function indicators and blood lipid parameters in reserpine model mice ( n≥5)

[0119] Group normal Model Imipramine 30 mg / kg ICCB35mg / kg AST(U / L) 152.20±24.35 <![CDATA[255.97±108.50 ## ]]> <![CDATA[167.16±53.17 * ]]> <![CDATA[172.01±37.65 * ]]> ALT(U / L) 49.44±17.26 <![CDATA[110.05±62.34 ## ]]> <![CDATA[51.85±13.62 ** ]]> <![CDATA[53.44±19.06 ** ]]> ALP(U / L) 153.01±30.10 <![CDATA[115.49±32.17 ## ]]> <![CDATA[149.95±33.74 ** ]]> <![CDATA[151.52±32.22 ** ]]> CHOL(mmoL / L) 4.30±0.69 <![CDATA[3.56±0.91 # ]]> <![CDATA[4.27±0.47 * ]]> <![CDATA[4.21±0.40 * ]]> HDL-C (mmoL / L) 2.37±0.17 <![CDATA[1.85±0.43 ## ]]> <![CDATA[2.33±0.28 * ]]> <![CDATA[2.31±0.31 * ]]> LDL-C (mmoL / L) 0.41±0.07 <![CDATA[0.27±012 # ]]> <![CDATA[0.42±0.09 ** ]]> <![CDATA[0.41±0.04 ** ]]> TG (mmoL / L) 1.73±0.44 <![CDATA[0.99±0.81 # ]]> <![CDATA[1.68±0.50 * ]]> <![CDATA[1.71±0.45 * ]]>

[0120] Note: Compared with the model group * P<0.05, ** P<0.01; compared with the normal group, # P<0.05, ## P<0.01

[0121] From Table 5 and Figures 9 to 15 It was found that compared with the normal group mice, the AST and ALT levels in the serum of the model group mice were significantly increased (P<0.01); the ALP, CHOL, HDL-C, LDL-C and TG levels were significantly decreased (P<0.05, P<0.01). After ICCB and imipramine administration, the AST and ALT levels of mice were significantly downregulated compared with the model group (P<0.05, P<0.01); the ALP, CHOL, HDL-C, LDL-C and TG levels were significantly increased (P<0.05, P<0.01). This shows that ICCB has a certain protective effect on the liver function and lipid metabolism function of the reserpine mouse model.

[0122] Example 2

[0123] The therapeutic effect of in vitro cultured bezoar on depression model induced by altered environmental conditions

[0124] 1. Construction of the Unpredictable Chronic Stress Model (CUMS)

[0125] CUMS modeling process: Each modeling mouse was randomly given 1-2 stimulations every day, including flashing strong light irradiation overnight, 40℃+100rmp / min shock for 1h, restraint for 1h, white noise for 8h, tail suspension for 5min, ice water swimming for 5min, water or food deprivation for 12h, wet dirty bedding overnight (water / bedding=100mL / 50g), 45° tilted cage for 24h, etc., and maintained for 63 days.

[0126] After one week of adaptive feeding, 80 mice were randomly divided into two groups: a normal group (10 mice) and a modeling group (70 mice). The normal group was fed with conventional feeding; the modeling group was subjected to CUMS modeling, and the CUMS modeling process was as above.

[0127] 49 days after modeling, the tail suspension test and forced swimming test were used to screen the mice with successful modeling, and 50 mice were regrouped, with 10 mice in each group, for subsequent experiments. Model group: CUMS continued to stimulate for 14 days, and normal saline was gavaged from 50 to 63 days; imipramine group: CUMS continued to stimulate for 14 days, and 30 mg / kg / d imipramine was gavaged from 50 to 63 days; low-dose ICCB group: CUMS continued to stimulate for 14 days, and 10 mg / kg / d ICCB was gavaged from 50 to 63 days; medium-dose ICCB group: CUMS continued to stimulate for 14 days, and 35 mg / kg / d ICCB was gavaged from 50 to 63 days; high-dose ICCB group: CUMS continued to stimulate for 14 days, and 60 mg / kg / d ICCB was gavaged from 50 to 63 days. The ICCB solvent was 0.5% sodium carboxymethylcellulose solution.

[0128] At the same time, the normal group was gavaged with normal saline on days 50 to 63.

[0129] The normal group and the model group were given an equal amount of saline by intragastric administration. The volume of saline in the normal group, the model group, the imipramine solution, and the ICCB solution were all converted according to the volume of 20 g / 0.3 mL.

[0130] 2. Detection of changes in depressive-like behavior in the CUMS mouse model after ICCB administration

[0131] Within 3 days after the CUMS model was established, the mice were subjected to tail suspension, forced swimming and sugar water preference tests using the same methods as in Example 1.

[0132] The effects of ICCB on the sugar water preference index, tail suspension immobility time and swimming immobility time of the CUMS depression mouse model are shown in Table 6 and Figures 16 to 18 shown. Fig.16 The effect of ICCB on the sugar preference index of CUMS depression mouse model; Fig.17 The effect of ICCB on the tail suspension immobility time in the CUMS depression mouse model; Fig.18 The effect of ICCB on the swimming immobility time in the CUMS depression mouse model.

[0133] Table 6 Effects of ICCB on sugar preference index, tail suspension immobility time and swimming immobility time in CUMS depression mouse model ( n=10)

[0134] Group Sugar water preference index (%) Tail suspension immobility time (s) Swimming immobility time (s) normal 84.72±7.54 121.80±22.67 203.31±25.54 Model <![CDATA[68.37±6.11 ### ]]> <![CDATA[220.20±32.80 ### ]]> <![CDATA[263.26±17.10 ### ]]> Imipramine 30 mg / kg <![CDATA[78.70±9.32 ** ]]> <![CDATA[164.50±37.34 ** ]]> <![CDATA[237.75±16.22 ** ]]> ICCB10mg / kg <![CDATA[73.91±5.19 * ]]> 211.50±35.25 255.53±10.28 ICCB35mg / kg <![CDATA[77.69±6.03 ** ]]> <![CDATA[176.50±35.35 * ]]> <![CDATA[236.06±13.86 ** ]]> ICCB 60mg / kg <![CDATA[75.79±7.53 * ]]> <![CDATA[180.80±45.25 * ]]> <![CDATA[243.56±14.76 * ]]>

[0135] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the normal group, ### P<0.001.

[0136] From Table 6 and Figures 16 to 18 It was found that compared with the normal group mice, the sugar water preference index, tail suspension immobility time and swimming immobility time of the model group mice were significantly changed (P<0.001). Compared with the model group, the sugar water preference index of the mice in the imipramine group was significantly increased (P<0.01), and the low, medium and high dose ICCB groups were significantly increased (P<0.05, P<0.01); at the same time, the tail suspension and swimming immobility time of the mice in the imipramine group and the medium and high dose groups were significantly reduced (P<0.05, P<0.01). The behavioral results showed that ICCB can effectively improve the depressive-like behavior of CUMS mice.

[0137] 3. Protective effect of ICCB on damaged hippocampal neurons in CUMS model mice

[0138] After the behavioral test, brain tissues of mice in the normal group, model group, imipramine group, and low-, medium-, and high-dose ICCB groups were taken for HE staining, and the specific method was the same as in Example 1.

[0139] Effects of ICCB on the damage of CA1, CA3 and DG regions of the hippocampus in the CUMS depression mouse model Fig.19 shown. Fig.19 Middle scale bar = 100 μm.

[0140] Depend on Fig.19It was found that compared with the normal group mice, there were more damaged cells in the CA1, CA3 and DG regions of the hippocampus of the model group mice, which were manifested as dark staining of the nucleus, shrinkage and deformation of the cells, and blurred boundaries between the nucleus and cytoplasm; at the same time, the overall number of neurons in the CA1 and CA3 regions of the model group decreased, and the arrangement was sparse. Compared with the model group, the number of damaged cells in each division of the hippocampus of the mice in the imipramine group, the medium-dose group and the high-dose group decreased, and the overall cell morphology was full and tightly arranged. This shows that ICCB has a certain protective effect on the hippocampal neurons of CUMS mice.

[0141] 4. Effects of ICCB on liver function and blood lipids in CUMS depression mouse model detected by blood analyzer

[0142] Another group of mice were selected and CUMS modeling was performed as above. After drug administration, blood was collected from the normal group, model group, imipramine group, and low-, medium-, and high-dose ICCB groups using the same method as in Example 1.

[0143] The effects of ICCB on the changes of liver function indexes and blood lipid parameters in CUMS model mice are shown in Table 7 and Figures 20 to 26 As shown. Fig. 20 The effect of ICCB on AST in CUMS model mice; Fig.21 The effect of ICCB on ALT in CUMS model mice; Fig. 22 The effect of ICCB on ALP in CUMS model mice; Fig.23 The effect of ICCB on CHOL in CUMS model mice; Fig.24 The effect of ICCB on HDL-C in CUMS model mice; Fig.25 The effect of ICCB on LDL-C in CUMS model mice; Fig.26 The effect of ICCB on TG in CUMS model mice.

[0144] Table 7 Effects of ICCB on changes in liver function indicators and blood lipid parameters in CUMS model mice ( n≥5)

[0145]

[0146] Note: Compared with the model group * P<0.05, ** P<0.01; compared with the normal group, # P<0.05, ## P<0.01.

[0147] From Table 7 and Figures 20 to 26It was found that compared with the normal group mice, the CUMS model group mice had significantly increased serum AST, ALT, ALP and TG levels (P<0.05, P<0.01); CHOL, LDL-C and HDL-C levels were significantly decreased (P<0.01). After ICCB and imipramine administration, the AST, ALT, ALP and TG levels of mice were significantly downregulated compared with the model group (P<0.05); CHOL, HDL-C and LDL-C levels were significantly increased (P<0.05, P<0.01). This indicates that ICCB has a certain protective effect on the liver function and lipid metabolism function of the CUMS mouse model.

[0148] In summary, by using the reserpine and CUMS depression mouse models, ICCB can significantly improve the depressive-like manifestations of the two animal models of reserpine and CUMS, proving for the first time that ICCB has antidepressant effects, and at the same time effectively inhibits the pathological damage of the hippocampus of mice after modeling. ICCB can significantly restore various indicators of liver function in model mice, especially the regulatory effect of ICCB on blood lipid levels that are significantly related to liver metabolism. This shows that ICCB not only improves the depressive-like manifestations and pathological damage of the two animal models of reserpine and CUMS, but also has a certain protective effect on the downregulated liver function and blood lipid metabolism function after modeling.

[0149] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Bezoar is used in the preparation of antidepressant products.

2. The application according to claim 1, characterized in that: The application includes any one or more of the following (1) to (6): (1) Improve weight changes; (2) Improve body temperature changes; (3) Improve behavioral changes; (4) inhibiting hippocampal neuron damage; (5) Improve liver function; (6) Improve lipid metabolism function.

3. The application according to claim 2, characterized in that: The inhibiting of hippocampal neuron damage includes: reducing the number of damaged cells and / or preventing neuron synapse damage.

4. The application according to claim 2, characterized in that: The improvement of liver function includes: any one or more of inhibiting the increase of AST, inhibiting the increase of ALT and inhibiting the decrease of ALP.

5. The application according to claim 2, characterized in that: The improvement of lipid metabolism function includes: significantly increasing the content of any one or more of CHOL, HDL-C, LDL-C and TG.

6. The use according to claim 1, characterized in that: The depression includes drug-induced depression and / or depression induced by changes in environmental conditions.

7. The application according to claim 1, characterized in that: The bezoar includes bezoar cultivated in vitro.

8. The use according to claim 1, characterized in that: The products include pharmaceutical products.

9. The use according to claim 8, characterized in that: The medicine also includes auxiliary materials.

10. The use according to claim 9, characterized in that: The excipients include pharmaceutically acceptable excipients.

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