Natural product probiotic preparation with depression improving effect and application of natural product probiotic preparation

Abstract

The invention relates to the field of probiotic preparations, and provides a natural product probiotic preparation with depression improving effect in order to solve the problem that in the prior art, single probiotics are used for intervening or treating depressive mood, and the effect is limited, the natural product probiotic preparation comprises lactococcus lactis WHH2078 and saffron crocus; the microorganism classification name of the lactococcus lactis WHH2078 is lactococcus lactis, the strain is preserved in the China General Microbiological Culture Collection Center on February 25, 2021, the preservation address is No.3, No.1 yard, Beichen West Road, Chaoyang District, Beijing, China, and the preservation number is CGMCC NO.21831. The invention further discloses a preparation method of the lactococcus lactis WHH2078. The natural product probiotic preparation can relieve depression / anxiety-like behaviors, callback the level of 5-hydroxytryptamine in the brain-haihorse area, and play a role in improving depression.

Description

Technical Field

[0001] The present invention relates to the field of probiotic preparations, and in particular to a natural product probiotic preparation having the effect of improving depression and an application thereof. Background Art

[0002] Depression is one of the most common mental disorders, characterized by persistent low mood, often accompanied by multiple dimensions of cognitive, behavioral, and physiological changes. Depression is a complex pathophysiological process involving the interaction of multiple factors, such as genetic susceptibility and environmental triggers, but its pathogenesis is not yet fully understood. Depending on the severity and classification of depressive episodes, treatment strategies may include psychotherapy techniques (such as cognitive behavioral therapy or interpersonal psychotherapy) and drug interventions (such as selective serotonin reuptake inhibitors (SSRIs) and tricyclic antidepressants (TCAs)). However, in many cases, drug therapy cannot achieve the desired effect and is accompanied by various adverse reactions, such as weight gain, hypotension, and anticholinergic effects. Therefore, it is of great significance to find a healthy and effective intervention or treatment method.

[0003] In recent years, a large number of literature reports have shown that probiotics have shown good intervention or treatment effects in relieving mood disorders such as anxiety and depression. In animal models, it was found that the intake of specific probiotic strains can reduce anxiety and depressive-like behaviors. Patent CN202110743748.0 authorizes a strain of Lactococcus lactis WHH2078 WHH2078 with the effect of improving depression. This strain has good adhesion properties, acid and bile resistance, and intestinal barrier protection functions. It can significantly lower serum corticosterone levels, relieve depression / anxiety-like behaviors by adjusting the body's depression-related neural factors, and has a significant effect on improving depression. The effect of a single probiotic in improving depression is relatively limited, and no products have been found that combine probiotics with natural products to intervene or treat depression. Summary of the invention

[0004] In order to overcome the problem that a single probiotic is used to intervene or treat depression in the prior art, and its effect is relatively limited, the present invention provides a natural product probiotic preparation with an effect of improving depression, wherein the natural product probiotic preparation combines Lactococcus lactis WHH2078 with saffron, and can relieve depression / anxiety-like behaviors, adjust the level of 5-hydroxytryptamine in the hippocampus, and play a role in improving depression. The present invention also provides an application of a natural product probiotic preparation with an effect of improving depression.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions: A natural product probiotic preparation having an effect of improving depression, comprising Lactococcus lactis WHH2078 and saffron; The microbial classification of Lactococcus lactis WHH2078 is named Lactococcus lactis. The strain was deposited in the General Microbiology Center of China Culture Collection Administration on February 25, 2021. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, and the deposit number is CGMCC NO.21831.

[0006] Some plants or their natural products, such as saffron, green tea and phosphatidylserine, are well-known for their small side effects and the effect of relieving depression. Saffron is a bulbous herb of the genus Crocus in the family Iridaceae. Its medicinal part is the dried stigma, which has the effects of promoting blood circulation and removing blood stasis, relieving depression and calming the nerves. Modern research shows that saffron contains a large amount of bioactive substances, mainly including crocin, crocin and safranal, which play an important role in the treatment of anxiety and depression, and have good safety and tolerance. Animal experiments have confirmed that saffron effectively relieves depression in mice by improving the 5-hydroxytryptamine system and dopamine system of depressed mice. The inventor found that Lactococcus lactis WHH2078 combined with saffron has the function of improving depression, and the compounding of the two has a synergistic effect.

[0007] Preferably, the Lactococcus lactis WHH2078 in the natural product probiotic preparation is in the form of bacterial liquid or freeze-dried bacterial powder.

[0008] Preferably, the freeze-dried bacterial powder further comprises a protective agent, which comprises skim milk, sucrose and monosodium glutamate.

[0009] Preferably, in the protective agent, the mass ratio of skim milk, sucrose and monosodium glutamate is (0.25-0.75):(0.1-0.5):(0.05-0.2).

[0010] Preferably, the preparation method of the freeze-dried bacterial powder is as follows: inoculating the activated Lactococcus lactis WHH2078 into an expansion culture medium for expansion culture; the culture medium is M17 culture medium, the inoculation amount of the expansion culture is 1-4%, the temperature is 37-43°C, the pH value is 5.5-6.5, and the time is 12-16h; nitrogen is passed through, and after the expansion culture is completed, the culture solution is centrifuged to remove the supernatant and collect the precipitate, and finally the precipitate is mixed with a protective agent and freeze-dried to prepare the freeze-dried bacterial powder.

[0011] Preferably, the dosage ratio of Lactococcus lactis WHH2078 to saffron in the natural product probiotic preparation is ((1-9)×10 10 )CFU: (10~90)mg.

[0012] The dosage of Lactococcus lactis WHH2078 in natural probiotic preparations is (1-9)×10 10CFU / person / day, the saffron dosage is 10-90 mg / person / day.

[0013] Preferably, the natural product probiotic preparation further comprises an excipient and / or a diluent.

[0014] Preferably, the excipient and / or diluent is food, medicine or health product.

[0015] The use of the above-mentioned natural product probiotic preparation with the effect of improving depression in the preparation of products with the effect of preventing and / or treating depression.

[0016] Preferably, the product is a tablet, a capsule or a powder.

[0017] Preferably, the product contains dairy products or fermented fruits and vegetables.

[0018] The above products are in oral form and are used to improve depression and relieve anxiety.

[0019] Therefore, the present invention has the following beneficial effects: (1) The natural product probiotic preparation WHHMOOD provided by the present invention is a combination of Lactococcus lactis WHH2078 and saffron, and the components interact synergistically to improve depression; (2) The natural product probiotic preparation WHHMOOD provided by the present invention is a combination of Lactococcus lactis WHH2078 and saffron, and is a natural, safe preparation that can effectively improve depression; (3) The natural product probiotic preparation WHHMOOD provided by the present invention can effectively alleviate the depression / anxiety-like behavior of depressed mice; (4) The natural product probiotic preparation WHHMOOD provided by the present invention can effectively alleviate the depression / anxiety-like behavior of depressed mice mainly by adjusting the central serotonin level. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a mouse trajectory diagram of the open field test of WHHMOOD of the present invention. DETAILED DESCRIPTION

[0021] The present invention is further described below in conjunction with specific implementation methods and drawings.

[0022] In the following specific implementation method, Saffron was purchased from Guangdong Qingyunshan Pharmaceutical Co., Ltd.; L-theanine was purchased from Tiancao Biotechnology Co., Ltd.; Phosphatidylserine was purchased from Cargill Japan Co., Ltd.; Lactococcus lactis WHH2078 was deposited at the General Microbiology Center of China Culture Collection Administration on February 25, 2021, at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, with the microbial collection number CGMCCNO.21831. It was isolated from yogurt and butter collected from rural areas in Shigatse City, Tibet Autonomous Region, my country; the morphological characteristics of Lactococcus lactis WHH2078 are: the growth form in M17 agar medium is milky white colonies, which are opaque, round, smooth and moist on the surface, with neat edges and a convex center. Gram staining is typically positive. Under a microscope, the cells are spherical, without flagella, do not produce spores, and do not move; the optimal growth temperature is 37°C, it is facultative anaerobic, and grows in M17 culture medium.

[0023] In the following specific implementation method, the freeze-dried bacterial powder of Lactococcus lactis WHH2078 is prepared by the following preparation method: inoculating the activated Lactococcus lactis WHH2078 into an expansion culture medium for expansion culture; the culture medium is M17 culture medium, the inoculation amount of the expansion culture is 2%, the temperature is 37°C, the pH value is 5.8, and the time is 15 hours; nitrogen is passed through, and after the expansion culture is completed, the culture solution is centrifuged to remove the supernatant and the precipitate is collected, and finally the precipitate is mixed with a protective agent and freeze-dried to prepare the freeze-dried bacterial powder; the protective agent includes skim milk, sucrose and monosodium glutamate in a mass ratio of 0.5:0.3:0.1.

[0024] Example 1 A natural product probiotic preparation with the function of improving depression, the natural product probiotic preparation is recorded as WHHMOOD, and the components are freeze-dried bacterial powder of Lactococcus lactis WHH2078 and saffron, wherein the dosage of Lactococcus lactis WHH2078 is 3×10 10 CFU / person / day, the dose of saffron is 30mg / person / day.

[0025] Comparative Example 1 A natural product probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078 and L-theanine, wherein the dosage of Lactococcus lactis WHH2078 is the same as that in Example 1, which is 3×10 10 CFU / person / day, the dosage of L-theanine is 18mg / person / day.

[0026] Comparative Example 2 A natural product probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078 and phosphatidylserine, wherein the dosage of Lactococcus lactis WHH2078 is the same as that in Example 1, which is 3×10 10 CFU / person / day, the dose of phosphatidylserine is 300 mg / person / day.

[0027] Comparative Example 3 A probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078, wherein the dosage of Lactococcus lactis WHH2078 is 3×10 10 CFU / person / day.

[0028] Comparative Example 4 A natural product with the function of improving depression, comprising saffron, and a dosage of saffron of 30 mg / person / day.

[0029] The depression-improving effects of the above-mentioned Examples and Comparative Examples were evaluated.

[0030] 1. Establishment of Depression Mouse Model The present invention uses healthy SPF male BALB / c mice (4-5 weeks old, 22±0.2g) and raises them in a clean environment with a room temperature of 26±0.5℃, a humidity of 50-60%, and a light-dark alternation time of 12 / 12h. A chronic unpredictable mild depression mouse model (CUMS) is established. The depression-inducing projects of CUMS mice include: restraint for 6h, swimming in 4℃ ice water for 1min, fasting for 24h, water deprivation for 24h, humid environment for 24h, and bedding-free environment for 24h, etc. The mice in the experimental group randomly perform one project every day.

[0031] 2. The experimental mice were divided into 8 groups, each with 10 mice, namely: control group, depression group, drug group, WHHMOOD group, comparative group 1, comparative group 2, comparative group 3, and comparative group 4. The normal group consisted of normal mice, which were gavaged with 200 μL of sterile saline every day; the depression group consisted of CUMS depressed mice, which were gavaged with 200 μL of sterile saline every day; the drug group consisted of CUMS depressed mice, which were gavaged with 200 μL of sterile fluoxetine solution (10 mg / kg BW) every day; the WHHMOOD group consisted of CUMS depressed mice, which were gavaged with 200 μL of WHHMOOD preparation (Lactococcus lactis WHH2078 5×10 9 CFU / kg / d and saffron 5 mg / kg / d, the conversion method is shown in Table 1); Comparative Example 1 group was CUMS depression mice, which were gavaged with 200 μL of the comparative example 1 preparation (Lactococcus lactis WHH2078 5×10 9 CFU / kg / d and L-theanine 3mg / kg / d); Comparative Example 2 group consisted of CUMS depression mice, which were intragastrically administered with 200 μL of the comparative example 2 preparation (Lactococcus lactis WHH2078 5×10 9 CFU / kg / d and phosphatidylserine 50mg / kg / d; Comparative Example 3 group was CUMS depression mice, which were gavaged with 200μL of the comparative example 3 preparation (Lactococcus lactis WHH2078 5×10 9CFU / kg / d); Comparative Example 4 group consisted of CUMS depression mice, which were gavaged with 200 μL of the comparative example 4 preparation (saffron 5 mg / kg / d) every day.

[0032] Table 1. Conversion table of oral gavage volume for each group of mice Group Mouse gavage dose (mg(CFU) / kg / d) Recommended daily dose for human body (mg(CFU) / person / day) Saffron 5 30 L-Theanine 3 18 Phosphatidylserine 50 300 WHH2078 <![CDATA[5×10 9 ]]> <![CDATA[3×10 10 ]]> Note: Recommended daily dose for human: (mouse gavage dose / 10)×60kg.

[0033] After 28 days of the experiment, the experimental mice were subjected to behavioral tests. The depressive-like behavior of the mice was evaluated by the tail suspension test and the forced swimming test, and the anxiety-like behavior of the mice was evaluated by the open field test. After the behavioral test, the mice were killed by dislocation of the neck, and the hippocampal tissue of the experimental mice was taken. After homogenization, the concentration of 5-hydroxytryptamine in the hippocampus was detected using an ELISA kit. The statistical results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and P < 0.05 indicated that the difference was statistically significant.

[0034] The results of the depressive-like behaviors of the experimental mice are shown in Table 2.

[0035] Table 2 Mouse behavioral results Group Tail suspension test immobility time (s) Forced swimming test immobility time (s) Central exploration time in open field test (s) Control group <![CDATA[36.83±5.14 b ]]> <![CDATA[28.83±2.86 b ]]> <![CDATA[28.11±4.03 a ]]> Depression group <![CDATA[82.41±10.22 a ]]> <![CDATA[74.33±4.94 a ]]> <![CDATA[15.53±3.31 b ]]> Drug group <![CDATA[37.02±4.90 b ]]> <![CDATA[30.99±5.95 b ]]> <![CDATA[28.94±5.93 a ]]> WHHMOOD Group <![CDATA[30.36±2.11 c ]]> <![CDATA[23.47±5.24 c ]]> <![CDATA[30.43±6.65 a ]]> Comparative Example 1 <![CDATA[43.05±3.08 b ]]> <![CDATA[36.56±4.46 b ]]> <![CDATA[23.20±8.14 ab ]]> Comparative Example 2 <![CDATA[44.28±0.27 b ]]> <![CDATA[34.28±0.27 b ]]> <![CDATA[22.28±8.28 ab ]]> Comparative Example 3 <![CDATA[37.22±3.82 b ]]> <![CDATA[31.32±3.48 b ]]> <![CDATA[29.13±4.32 a ]]> Comparative Example 4 <![CDATA[40.48±2.38 b ]]> <![CDATA[34.55±1.43 b ]]> <![CDATA[23.49±6.43 ab ]]> a, b, c: significant difference, P<0.05.

[0036] The immobility time of the tail suspension test and the immobility time of the forced swimming test in the depression group mice were significantly higher than those in the control group mice (P<0.05), indicating that the modeling was successful. The treatment schemes of the drug group, WHHMOOD group, comparative example 1 group, comparative example 2 group, comparative example 3 group and comparative example 4 group in the present invention can significantly reduce the immobility time of the tail suspension test of the depressed mice (P<0.05), among which WHHMOOD has the best reduction effect, and the reduction range is 15% higher than that of comparative example 1, 16% higher than that of comparative example 2, 8% higher than that of comparative example 3, 12% higher than that of comparative example 4, and 8% higher than that of the drug group. The treatment schemes of the present invention, WHHMOOD, comparative example 1, comparative example 2, comparative example 3 and comparative example 4 can significantly reduce the immobility time of the forced swimming test of depressed mice (P<0.05), among which WHHMOOD has the best reduction effect, and the reduction amplitude is 17% higher than that of comparative example 1, 14% higher than that of comparative example 2, 10% higher than that of comparative example 3, 14% higher than that of comparative example 4, and 10% higher than that of the drug group. Therefore, the WHHMOOD combination provided by the present invention has the effect of relieving depression-like behavior, and is significantly better than the effects of fluoxetine, lactic acid such as coccus WHH2078 and L-theanine, lactic acid such as coccus WHH2078 and phosphatidylserine, lactic acid such as coccus WHH2078, and saffron in relieving depression-like behavior.

[0037] The results of the anxiety-like behaviors of the experimental mice are shown in Table 2 and Figure 1 As shown, the central exploration time of the open field test in the depression group mice was significantly lower than that in the control group mice (P<0.05), indicating that the depression mice were accompanied by anxiety-like behavior. The WHHMOOD treatment scheme in the present invention can significantly increase the central exploration time of the open field test in depression mice (P<0.05), and the increase is 45% higher than that of the effect of comparative example 1, 51% higher than that of comparative example 2, and 45% lower than that of comparative example 4, which is equivalent to the effect of the drug group (P>0.05). Therefore, the WHHMOOD combination provided by the present invention has the effect of relieving anxiety-like behavior, and is significantly better than the effects of lactic acid such as coccus WHH2078 and L-theanine, lactic acid such as coccus WHH2078 and phosphatidylserine and saffron in relieving anxiety-like behavior.

[0038] The 5-HT levels in the hippocampus of the experimental mice are shown in Table 3.

[0039] Table 3 Results of 5-HT detection in the hippocampus of mice Group Hippocampus 5-HT (μg / g) Control group <![CDATA[177.42±9.92 a ]]> Depression group <![CDATA[117.72±4.57 d ]]> Drug group <![CDATA[159.56±8.83 b ]]> WHHMOOD Group <![CDATA[163.29±8.50 b ]]> Comparative Example 1 <![CDATA[128.82±5.66 cd ]]> Comparative Example 2 <![CDATA[132.30±17.58 c ]]> Comparative Example 3 <![CDATA[150.02±5.92 bc ]]> Comparative Example 4 <![CDATA[140.67±8.44 bc ]]> a, b, c: significant difference, P<0.05.

[0040] The concentration of 5-hydroxytryptamine in the hippocampus of the depressed mice was significantly lower than that in the control group mice (P<0.05), indicating that the modeling was successful. In the present invention, the WHHMOOD treatment scheme can significantly increase the concentration of 5-hydroxytryptamine in the hippocampus of depressed mice (P<0.05), and the increase is 28% higher than the effect of comparative example 1, 25% higher than the effect of comparative example 2, 12% higher than the effect of comparative example 3, and 20% higher than the effect of comparative example 4, which is comparable to the effect of the drug group (P>0.05). Therefore, the WHHMOOD combination provided by the present invention has the ability to increase the level of 5-hydroxytryptamine in the hippocampus of depressed mice, and is significantly better than lactic acid such as coccus WHH2078 and L-theanine, lactic acid such as coccus WHH2078 and phosphatidylserine, lactic acid such as coccus WHH2078 and saffron in this respect.

[0041] Example 2 A natural product probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078 and saffron, wherein the ratio of freeze-dried bacterial powder of Lactococcus lactis WHH2078 to saffron is 2:1, wherein the dosage of Lactococcus lactis WHH2078 is 6×10 10 CFU / person / day, the dose of saffron is 30mg / person / day.

[0042] Example 3 A natural product probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078 and saffron, wherein the ratio of freeze-dried bacterial powder of Lactococcus lactis WHH2078 to saffron is 1:2, wherein the dosage of Lactococcus lactis WHH2078 is 3×10 10 CFU / person / day, the dose of saffron is 60mg / person / day.

[0043] The depression-improving effects of Examples 1 to 3 were evaluated using the same depression mouse model and assay method as above. The dosage conversion method for mice is the same as that in Table 1.

[0044] Table 4 Experimental results of Examples 1 to 3 Group Forced swimming test immobility time (s) Hippocampus 5-HT (μg / g) Control group <![CDATA[34.76±3.25 c ]]> <![CDATA[212.43±7.56 a ]]> Depression group <![CDATA[77.45±7.88 a ]]> <![CDATA[118.78±3.44 d ]]> Drug group <![CDATA[33.67±4.76 c ]]> <![CDATA[203.65±5.12 ab ]]> WHHMOOD Group <![CDATA[31.78±4.23 c ]]> <![CDATA[193.55±8.45 b ]]> Example 2 Group <![CDATA[39.90±4.11 b ]]> <![CDATA[149.45±3.22 c ]]> Example 3 Group <![CDATA[41.25±1.78 b ]]> <![CDATA[137.33±4.90 c ]]> a, b, c: significant difference, P<0.05.

[0045] The test results of different proportions are shown in Table 4. The forced swimming immobility time of the depressed mice was significantly higher than that of the control group mice, and the 5-hydroxytryptamine concentration in the hippocampus was significantly lower than that of the control group mice (P<0.05), indicating that the modeling was successful. The treatment schemes of the drug, WHHMOOD group, Example 2 group and Example 3 group in the present invention can significantly reduce the forced swimming immobility time of depressed mice (P<0.05), among which WHHMOOD has the best reduction effect, and the reduction amplitude is increased by 11% over the effect of Example 2, and the reduction amplitude is increased by 12% over the effect of Example 3, which is equivalent to the effect of the drug group (P>0.05). The drug, WHHMOOD, Example 2 and Example 3 treatment schemes in the present invention can significantly increase the 5-hydroxytryptamine concentration in the hippocampus of depressed mice (P<0.05), among which WHHMOOD has the best increase effect, and the increase amplitude is increased by 37% over the effect of Example 2, and the increase amplitude is increased by 47% over the effect of Example 3, which is equivalent to the effect of the drug group (P>0.05). Therefore, the WHHMOOD combination provided by the present invention (the ratio of lactic acid bacteria WHH2078 and saffron is 1:1) is significantly better than Example 2 (the ratio of lactic acid bacteria WHH2078 and saffron is 2:1) and Example 3 (the ratio of lactic acid bacteria WHH2078 and saffron is 1:2) in alleviating depression-like behavior and upregulating the level of 5-hydroxytryptamine in the hippocampus of depressed mice.

[0046] Example 4 A natural product probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078 and saffron, wherein the ratio of freeze-dried bacterial powder of Lactococcus lactis WHH2078 to saffron is 1:1, wherein the dosage of Lactococcus lactis WHH2078 is 1×10 10 CFU / person / day, the dose of saffron is 10mg / person / day.

[0047] Example 5 A natural product probiotic preparation with the function of improving depression, comprising freeze-dried bacterial powder of Lactococcus lactis WHH2078 and saffron, wherein the ratio of freeze-dried bacterial powder of Lactococcus lactis WHH2078 to saffron is 1:1, wherein the dosage of Lactococcus lactis WHH2078 is 9×10 10 CFU / person / day, the dose of saffron is 90mg / person / day.

[0048] The depression-improving effects of Examples 1 and 4-5 were evaluated using the same depression mouse model and assay method as above. The dosage conversion method for mice was the same as that in Table 1.

[0049] Table 5 Test results of Example 1, Example 4-5 Group Forced swimming test immobility time (s) Hippocampus 5-HT (μg / g) Control group <![CDATA[26.21±5.34 c ]]> <![CDATA[209.15±5.23 a ]]> Depression group <![CDATA[84.38±5.54 a ]]> <![CDATA[130.47±5.68 c ]]> Drug group <![CDATA[31.93±4.12 c ]]> <![CDATA[180.15±6.43 b ]]> WHHMOOD Group <![CDATA[28.34±4.56 c ]]> <![CDATA[178.34±4.46 b ]]> Example 4 Group <![CDATA[43.68±6.73 b ]]> <![CDATA[148.89±3.12 bc ]]> Example 5 Group <![CDATA[28.00±6.94 c ]]> <![CDATA[180.30±6.23 b ]]> a, b, c: significant difference, P<0.05.

[0050] The experimental results of different doses are shown in Table 5. The forced swimming immobility time of the depressed mice was significantly higher than that of the control group mice, and the 5-hydroxytryptamine concentration in the hippocampus was significantly lower than that of the control group mice (P<0.05), indicating that the modeling was successful. The drug, WHHMOOD, Example 4 and Example 5 treatment schemes of the present invention can significantly reduce the forced swimming immobility time of depressed mice (P<0.05), wherein the reduction effect of the WHHMOOD group and the Example 5 group is equivalent to the effect of the drug group (P>0.05), and is better than the effect of Example 4 (P<0.05). The drug, WHHMOOD, Example 4 and Example 5 treatment schemes of the present invention can significantly increase the 5-hydroxytryptamine concentration in the hippocampus of depressed mice (P<0.05), wherein the increase effect of WHHMOOD and Example 5 is equivalent to the effect of the drug group (P>0.05), and is better than the effect of Example 4 (P<0.05). Therefore, the WHHMOOD combination provided by the present invention, wherein the dosage ratio of Lactococcus lactis WHH2078 to saffron is ((1-9)×10 10 )CFU: (10~90)mg, has the effect of improving depression and shows dosage effect.

[0051] In summary, the natural product probiotic preparation of the present invention can significantly alleviate depression / anxiety-like behaviors and adjust the 5-hydroxytryptamine level in the hippocampus of the brain. It is a natural product probiotic preparation with the function of improving depression.

[0052] Example 6 A WHHMOOD solid beverage for improving depression, the formula of which is as follows: based on 100g of solid beverage: 5g of Lactococcus lactis WHH2078 freeze-dried powder, 2g of saffron extract, 38g of erythritol, 25g of strawberry powder, 15g of polydextrose powder, 14g of maltodextrin, and 1g of citric acid; The number of viable bacteria in the Lactococcus lactis WHH2078 freeze-dried powder is 2×10 11 CFU / g; The method for preparing the solid beverage comprises the following steps: (1) Weigh each raw material for later use; (2) mixing the raw materials except the Lactococcus lactis WHH2078 freeze-dried powder uniformly at a mixing speed of 30 rpm for a mixing time of 15 min to obtain a mixed semi-finished product; (3) boiling granulation of the mixed semi-finished product obtained in step (2); pure water is used as a binder in the boiling granulation process; during the boiling granulation, the inlet air temperature is 85° C., the exhaust air frequency is 80%, the material temperature is 50° C., the atomization pressure is 2.5 bar, and the speed of the spray slurry pump is 80 rpm; the product after boiling granulation is sieved, and the sieve is a 20-mesh standard sieve to obtain a boiling granulation semi-finished product; (4) mixing the boiling granulation semi-finished product obtained in step (3) and the freeze-dried powder of Lactococcus lactis WHH2078 uniformly at a mixing speed of 30 rpm for a mixing time of 15 min to obtain a total mixed semi-finished product; (5) packaging the total mixed semi-finished product obtained in step (4) using a stick pack filling machine, filling with nitrogen during packaging, with a residual oxygen content of 5%, and using an aluminum-plastic packaging material to obtain a weight loss powder based on lactobacillus; Steps (1) to (5) are all carried out in a constant temperature and humidity environment in a 100,000-grade GMP workshop, with a temperature of 25° C. and a humidity of 40%.

[0053] The obtained weight loss powder was tested for key indicators, and the test results are shown in Table 6.

[0054] Table 6 Key index test results Example 7 A WHHMOOD solid tablet for improving depression, the formula of which is as follows: based on 100g of solid tablets: 5g of Lactococcus lactis WHH2078 freeze-dried powder, 2g of saffron extract, 5g of blueberry powder, 86.95g of direct compression excipients, 0.05g of vitamin C and 1g of magnesium stearate; the number of viable bacteria in the Lactococcus lactis WHH2078 freeze-dried powder is 2×10 11 CFU / g; the direct compression excipient is direct compression maltitol; The method for preparing the solid tablet comprises the following steps: (1) Weigh each raw material according to the weight ratio for use; (2) Mixing 0.05 g of vitamin C and 1.95 g of direct compression maltitol uniformly to obtain a mixed small material; (3) 5 g of Lactococcus lactis WHH2078 freeze-dried powder, 2 g of saffron extract, 5 g of blueberry powder, and 85 g of direct compression sorbitol were mixed evenly with the mixed small materials to obtain a primary mixture; the mixing speed was 35 rpm and the mixing time was 20 min; (4) The obtained preliminary mixture was mixed evenly with 1 g of magnesium stearate to obtain a total mixed semi-finished product; the mixing speed was 25 rpm and the mixing time was 20 min; (5) tabletting the obtained total mixed semi-finished product using a tablet press to obtain tablets; the tableting pressure during the tableting process is 16 kN; (6) packaging the tablets obtained in the step with a packaging bottle to obtain a finished product; the packaging bottle used for packaging is a high-density polyethylene (HDPE) packaging bottle with a built-in desiccant; Among them, in steps (1) to (6), all operations are carried out in a constant temperature and humidity environment in a 100,000-grade GMP workshop, with a temperature of 18° C. and a humidity of 40%.

[0055] The obtained tablet finished product was tested for key indicators, and the test results are shown in Table 7.

[0056] Table 7 Key index test results

Claims

1. A natural product probiotic preparation having an effect of improving depression, characterized in that: Includes Lactococcus lactis WHH2078 and saffron; The microbial classification of Lactococcus lactis WHH2078 is named Lactococcus lactis Lactococcus lactis The strain was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on February 25, 2021. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, China, and the deposit number is CGMCC NO.21831.

2. A natural product probiotic preparation having an effect of improving depression according to claim 1, characterized in that: In the natural product probiotic preparation, Lactococcus lactis WHH2078 exists in the form of bacterial liquid or freeze-dried bacterial powder.

3. A natural product probiotic preparation having an effect of improving depression according to claim 2, characterized in that: The freeze-dried bacterial powder also includes a protective agent, which includes skim milk, sucrose and monosodium glutamate.

4. A natural product probiotic preparation having an effect of improving depression according to claim 3, characterized in that: In the protective agent, the mass ratio of skim milk, sucrose and monosodium glutamate is (0.25-0.75): (0.1-0.5): (0.05-0.2).

5. A natural product probiotic preparation having an effect of improving depression according to claim 1, 2, 3 or 4, characterized in that: The dosage ratio of Lactococcus lactis WHH2078 and saffron in the natural product probiotic preparation is ((1~9)×10 10 )CFU:(10~90)mg.

6. The natural product probiotic preparation having the effect of improving depression according to claim 1, characterized in that: The natural product probiotic preparation also includes excipients and / or diluents.

7. A natural product probiotic preparation having an effect of improving depression according to claim 6, characterized in that: The excipient and / or diluent is food, medicine or health product.

8. Use of the natural product probiotic preparation with depression-improving effect as claimed in any one of claims 1 to 7 in the preparation of a product with depression-preventing and / or treating effect.

9. The use according to claim 8, characterized in that: The product is in the form of tablets, capsules or powders.

10. The use according to claim 8, characterized in that: The product contains dairy products or fermented fruits and vegetables.

Images