CN117398417A - Application of lactobacillus paracasei E6 in preparation of antidepressant drugs - Google Patents
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- 241000186605 Lactobacillus paracasei Species 0.000 title claims abstract description 57
- 239000000935 antidepressant agent Substances 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 claims abstract description 40
- 210000005013 brain tissue Anatomy 0.000 claims abstract description 16
- 239000006041 probiotic Substances 0.000 claims abstract description 4
- 230000000529 probiotic effect Effects 0.000 claims abstract description 4
- 235000018291 probiotics Nutrition 0.000 claims abstract description 4
- 230000001580 bacterial effect Effects 0.000 claims description 13
- 229940005513 antidepressants Drugs 0.000 claims description 11
- 230000001430 anti-depressive effect Effects 0.000 claims description 8
- 239000000725 suspension Substances 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 241000252212 Danio rerio Species 0.000 abstract description 37
- 230000000694 effects Effects 0.000 abstract description 15
- DNXIKVLOVZVMQF-UHFFFAOYSA-N (3beta,16beta,17alpha,18beta,20alpha)-17-hydroxy-11-methoxy-18-[(3,4,5-trimethoxybenzoyl)oxy]-yohimban-16-carboxylic acid, methyl ester Natural products C1C2CN3CCC(C4=CC=C(OC)C=C4N4)=C4C3CC2C(C(=O)OC)C(O)C1OC(=O)C1=CC(OC)=C(OC)C(OC)=C1 DNXIKVLOVZVMQF-UHFFFAOYSA-N 0.000 abstract description 14
- LCQMZZCPPSWADO-UHFFFAOYSA-N Reserpilin Natural products COC(=O)C1COCC2CN3CCc4c([nH]c5cc(OC)c(OC)cc45)C3CC12 LCQMZZCPPSWADO-UHFFFAOYSA-N 0.000 abstract description 14
- QEVHRUUCFGRFIF-SFWBKIHZSA-N Reserpine Natural products O=C(OC)[C@@H]1[C@H](OC)[C@H](OC(=O)c2cc(OC)c(OC)c(OC)c2)C[C@H]2[C@@H]1C[C@H]1N(C2)CCc2c3c([nH]c12)cc(OC)cc3 QEVHRUUCFGRFIF-SFWBKIHZSA-N 0.000 abstract description 14
- BJOIZNZVOZKDIG-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C([C]5C=CC(OC)=CC5=N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 BJOIZNZVOZKDIG-MDEJGZGSSA-N 0.000 abstract description 14
- 229960003147 reserpine Drugs 0.000 abstract description 14
- MDMGHDFNKNZPAU-UHFFFAOYSA-N roserpine Natural products C1C2CN3CCC(C4=CC=C(OC)C=C4N4)=C4C3CC2C(OC(C)=O)C(OC)C1OC(=O)C1=CC(OC)=C(OC)C(OC)=C1 MDMGHDFNKNZPAU-UHFFFAOYSA-N 0.000 abstract description 14
- 238000000034 method Methods 0.000 abstract description 4
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- RTHCYVBBDHJXIQ-MRXNPFEDSA-N (R)-fluoxetine Chemical compound O([C@H](CCNC)C=1C=CC=CC=1)C1=CC=C(C(F)(F)F)C=C1 RTHCYVBBDHJXIQ-MRXNPFEDSA-N 0.000 description 10
- 229960002464 fluoxetine Drugs 0.000 description 10
- 239000013641 positive control Substances 0.000 description 8
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- 208000020401 Depressive disease Diseases 0.000 description 3
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- 238000008157 ELISA kit Methods 0.000 description 2
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- 229940079593 drug Drugs 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
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- 229940123685 Monoamine oxidase inhibitor Drugs 0.000 description 1
- 240000000907 Musa textilis Species 0.000 description 1
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- 206010042458 Suicidal ideation Diseases 0.000 description 1
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- 238000004458 analytical method Methods 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
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Abstract
The invention discloses application of lactobacillus paracasei E6 in preparation of antidepressant drugs, and belongs to the technical field of microorganisms. The application of the lactobacillus paracasei E6 in preparing antidepressant drugs disclosed by the invention has the advantages that the lactobacillus paracasei E6 can obviously improve the activity degree of zebra fish and the 5-HT concentration in brain tissue in a zebra fish depression model induced by reserpine, namely, the anti-depression effect is realized; the method provides theoretical reference and guiding basis for developing a probiotic preparation for treating depression by using lactobacillus paracasei E6.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application of lactobacillus paracasei E6 in preparation of antidepressant drugs.
Background
Depression is a common mental disorder characterized by a marked and persistent depression in the mood. Clinical manifestations include physical discomfort such as fatigue, insomnia, inappetence, hyposexuality, headache, dizziness, thirst, pharyngeal discomfort, chest distress, palpitation, gastralgia, abdominal distention, etc., and severe cases may have a feeling of self-intolerance, even suicidal tendency, in addition to mental symptoms such as mental depression, loss of interest, slow thinking, impaired self-assessment, etc. With the progress of society and the development of economy, the social competition is increased, the life rhythm is accelerated, the social status is changed rapidly, and the depression becomes a common disease and a high morbidity of the modern society, and the morbidity is rising rapidly.
The treatment of depression mainly comprises drug treatment, psychological treatment and physical treatment. Antidepressants are the main mode for treating depressive disorder at present, and have therapeutic effects on depressed mood and accompanying anxiety, tension and somatic symptoms, and the effective rate can reach 60% -70%. Currently common therapeutic agents include tricyclic and tetracyclic antidepressants, monoamine oxidase inhibitors, selective 5-hydroxytryptamine reuptake inhibitors, and the like; the adverse reaction is more after the treatment, and the side effect is large.
Thus, providing the use of lactobacillus paracasei E6 for the preparation of antidepressants is a problem to be solved by the person skilled in the art.
Disclosure of Invention
In view of this, the present invention provides the use of lactobacillus paracasei E6 in the preparation of antidepressants.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
use of lactobacillus paracasei E6 for the preparation of an antidepressant, said lactobacillus paracasei E6 having a preservation number of CGMCC No.21771 (see patent No. 202211604325.1).
Further, the lactobacillus paracasei E6 bacterial suspension is applied to the preparation of antidepressants.
Further, the lactobacillus paracasei E6 is applied to preparing the medicines for improving the 5-HT concentration in brain tissues.
Further, the lactobacillus paracasei E6 bacterial suspension is applied to preparing a medicament for improving the 5-HT concentration in brain tissues.
Further, an antidepressant product comprising said lactobacillus paracasei E6.
Further, the product is a probiotic preparation.
Compared with the prior art, the invention discloses application of the lactobacillus paracasei E6 in preparing antidepressant drugs, wherein the lactobacillus paracasei E6 is obtained by separating and screening faeces of long-life old people in the city of the abaca county in Guangdong province; the lactobacillus paracasei E6 can obviously improve the activity degree of zebra fish and the 5-HT concentration in brain tissue in a zebra fish depression induced by reserpine model, namely has the depression resisting effect; the method provides theoretical reference and guiding basis for developing a probiotic preparation for treating depression by using lactobacillus paracasei E6.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the effect of Lactobacillus paracasei E6 of the present invention on the range of motion of reserpine-induced zebra fish depression models;
FIG. 2 is a graph showing the effect of Lactobacillus paracasei E6 of the present invention on average motor speed in a reserpine-induced zebra fish depression model;
FIG. 3 is a graph showing the effect of Lactobacillus paracasei E6 of the present invention on resting time in a reserpine-induced zebra fish depression model;
FIG. 4 is a graph showing the effect of Lactobacillus paracasei E6 of the present invention on 5-HT in brain tissue of a model of reserpine-induced depression in zebra fish.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Reserpine was purchased from Sigma; fluoxetine was purchased from MedChemExpress corporation; zebra fish 5-hydroxytryptamine (5-HT) ELISA kit was purchased from DiXintai detection technologies (Beijing); lactobacillus paracasei 1.570 (CGMCC 1.570) was purchased from the chinese common microbiological bacterial collection center.
EXAMPLE 1 preparation of Lactobacillus paracasei E6 suspension (thallus)
Inoculating lactobacillus paracasei E6 after activation culture in MRS liquid culture medium, culturing at 37 ℃ for 24 hours, and centrifuging at 4 ℃ for 10 minutes at 6000r/min to obtain bacterial precipitate; after the bacterial cell precipitate is washed twice by PBS, the bacterial cell is resuspended by PBS, and the cell concentration is regulated to be 1 multiplied by 10 6 CFU/mL gave a bacterial suspension (cell).
EXAMPLE 2 preparation of Lactobacillus paracasei 1.570 suspension (thallus)
Inoculating lactobacillus paracasei 1.570 after activation culture in MRS liquid culture medium, culturing at 37 ℃ for 24 hours, and centrifuging at 4 ℃ for 10min at 6000r/min to obtain bacterial precipitate; after the bacterial cell precipitate is washed twice by PBS, the bacterial cell is resuspended by PBS, and the cell concentration is regulated to be 1 multiplied by 10 6 CFU/mL gave a bacterial suspension (cell).
Example 3 Effect of Lactobacillus paracasei E6 on movement of the zebra fish depression model
Healthy wild-type AB-line zebra fish that developed to 3dpf (days post fertilization) were selected and placed in 96-well cell culture plates. Experiment setting normal group, model group (reserpine), positive control group (fluoxetine), lactobacillus paracasei 1.570 intervention group (1×10) 6 CFU/mL), lactobacillus paracasei E6 intervention group (1X 10) 6 CFU/mL), 20 duplicate wells were placed per group, 1 fish per well. (1) 200 mu L of PBS is added into each hole of a normal group, 200 mu L of reserpine solution (2 mg/L) is added into each hole of a model group, a positive control group, a Lactobacillus paracasei 1.570 intervention group and a Lactobacillus paracasei E6 intervention group, after incubation for 1h at 28 ℃, the solution is discarded, and PBS is added into both the normal group and the model group; the positive control group was added with fluoxetine solution (4. Mu. Mol/L); lactobacillus paracasei 1.570 intervention group (1×10) 6 CFU/mL) was added 1X 10 6 CFU/mL Lactobacillus paracasei 1.570; cheese milkBacillus E6 intervention group (1X 10) 6 CFU/mL) was added 1X 10 6 CFU/mL Lactobacillus paracasei E6, 200. Mu.L per well, were incubated at 28 ℃. (2) Repeating the step (1) after every 24 hours, replacing the new solution, incubating for 72 hours at 28 ℃, placing the 96-well cell culture plate in a zebra fish behavior analysis system (EthoVision XT 10.4, nodas information technology Co., ltd.) for tracking the track, turning off a light source to adapt the zebra fish to the environment for 5 minutes, and then starting behavior track analysis software to collect the behavior track of the zebra fish in the dark environment for 5 minutes. And counting the movement distance, the average movement speed and the resting time of the zebra fish.
SPSS19.0 software was used to statistically process the data, experimental data were all expressed as x+ -SEM data, analyzed by T-test, compared to normal group: ### P<0.005, compared to model group: * P:<0.005。
the results are shown in FIGS. 1-3; as can be seen from fig. 1, 2 and 3, compared with the normal group (movement distance: 49.98 ±2.25cm; average movement speed: 1.67±0.07mm/s; resting time: 159.20 ±6.40 s), the movement distance (29.59±2.31 cm) and average movement speed (0.99±0.08 mm/s) of the zebra fish of the model group were significantly reduced (p < 0.005), and the resting time (250.44 ±4.83 s) was significantly increased (p < 0.005), indicating that the current zebra fish depression model construction was successful.
Compared with the model group (movement distance: 29.59+ -2.31 cm; average movement speed: 0.99+ -0.08 mm/s; resting time: 250.44 + -4.83 s), the movement distance (45.45+ -2.56 cm) and average movement speed (1.51+ -0.09 mm/s) of the zebra fish of the positive control group (fluoxetine) were significantly improved (p)<0.005 And a significant reduction in the rest time (188.92.+ -. 7.47 s) (p)<0.005 The fluoxetine is shown to obviously improve the activity degree of the zebra fish in a zebra fish depression model induced by reserpine. Thus, fluoxetine has depression relieving effects, consistent with clinical results. Lactobacillus paracasei 1.570 intervention group (1×10) 6 CFU/mL), the motion distance, average motion speed, and resting time of the zebra fish were 34.42 ±2.73cm, 1.15±0.09mm/s, 240.53 ±4.11s, respectively, and the motion distance of the zebra fish was determined with the model group (motion distance: 29.59+ -2.31 cm; average movement speed: 0.99+/-0.08 mm/s; rest time: 250.44.+ -. 4.83 s) no significant difference (P)>0.05). In addition, withCompared with the model group (movement distance: 29.59.+ -. 2.31cm; average movement speed: 0.99.+ -. 0.08mm/s; resting time: 250.44.+ -. 4.83 s), lactobacillus paracasei E6 intervention group (1X 10) 6 The movement distance (42.52 +/-1.68 cm) and the average movement speed (1.42+/-0.06 mm/s) of the zebra fish (CFU/mL) are obviously improved (p)<0.005 And a significant reduction in the rest time (208.58.+ -. 9.05 s) (p<0.005). Therefore, the results show that the lactobacillus paracasei E6 can obviously improve the activity degree of the zebra fish in a reserpine-induced zebra fish depression model, namely has the effect of relieving depression.
Example 4 Effect of Lactobacillus paracasei E6 on 5-hydroxytryptamine (5-HT) in brain tissue of a model of zebra fish depression
Healthy wild-type AB-line zebra fish that developed to 3dpf (days post fertilization) were selected and placed in 6-well cell culture plates. Experiment setting normal group, model group (reserpine), positive control group (fluoxetine), lactobacillus paracasei 1.570 intervention group (1×10) 6 CFU/mL), lactobacillus paracasei E6 intervention group (1X 10) 6 CFU/mL), 6 duplicate wells were placed per group, 40 fish per well. (1) 5mL of PBS is added into each hole of a normal group, 5mL of reserpine solution (2 mg/L) is added into each hole of a model group, a positive control group, a Lactobacillus paracasei 1.570 intervention group and a Lactobacillus paracasei E6 intervention group, after incubation for 1h at 28 ℃, PBS is added into the normal group and the model group; the positive control group was added with fluoxetine solution (4. Mu. Mol/L); lactobacillus paracasei 1.570 intervention group (1×10) 6 CFU/mL) was added 1X 10 6 CFU/mL Lactobacillus paracasei 1.570; lactobacillus paracasei E6 intervention group (1X 10) 6 CFU/mL) was added 1X 10 6 CFU/mL Lactobacillus paracasei E6, 5mL per well, were incubated at 28 ℃. (2) Repeating the step (1) after every 24 hours to replace a new solution, after incubating for 72 hours at 28 ℃, collecting the zebra fish brain tissue, flushing with PBS, taking 50mg of the brain tissue, placing into a centrifuge tube, adding 0.5mL of PBS, homogenizing and crushing the zebra fish brain tissue by using an S-18KS handheld micro-electric tissue homogenizer until no obvious tissue fragments exist, centrifuging for 15 minutes at 4 ℃, and collecting each group of supernatant. The 5-HT concentration of each group was measured using a zebra fish 5-hydroxytryptamine (5-HT) ELISA kit.
SPSS19.0 software is adopted to statistically process data, and experimental data are all expressed by x (+ -)SEM data indicated that were analyzed by T-test, compared to normal group: ### P<0.005, compared to model group: * P:<0.005。
the results are shown in FIG. 4; as can be seen from FIG. 4, the concentration of 5-HT (17.33+ -1.09 pg/mL) in the brain tissue of the zebra fish in the model group was significantly reduced (p < 0.005) compared to the normal group (42.40+ -2.93 pg/mL), indicating that the current zebra fish depression model was successfully constructed.
The concentration of 5-HT in the brain tissue of zebra fish in the positive control group (fluoxetine) was 36.13+ -2.54 pg/mL, which was significantly different from that in the model group (17.33+ -1.09 pg/m) (P<0.005 Indicated that fluoxetine has antidepressant effect and is consistent with clinical results. Lactobacillus paracasei 1.570 intervention group (1×10) 6 CFU/mL) the concentration of 5-HT in the brain tissue of zebra fish was 19.62+ -1.78 pg/mL, with no significant differences (P) compared to the model group (17.33+ -1.09 pg/m)>0.05). While Lactobacillus paracasei E6 intervention group (1X 10) 6 The concentration of 5-HT in brain tissue of the zebra fish was 32.20+ -2.32 pg/mL, with significant poorer variability (P) compared to the model group (17.33+ -1.09 pg/m)<0.005). Therefore, the results show that the lactobacillus paracasei E6 can obviously improve the 5-HT concentration in the brain tissue of the zebra fish in a reserpine-induced zebra fish depression model, namely has an antidepressant effect.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. The application of lactobacillus paracasei E6 in preparing antidepressant is characterized in that the preservation number of the lactobacillus paracasei E6 is CGMCC No.21771.
2. Use of a bacterial suspension of lactobacillus paracasei E6 as defined in claim 1 for the preparation of an antidepressant.
3. Use of lactobacillus paracasei E6 as claimed in claim 1 for the manufacture of a medicament for increasing the concentration of 5-HT in brain tissue.
4. Use of a bacterial suspension of lactobacillus paracasei E6 as defined in claim 1 for the preparation of a medicament for increasing the concentration of 5-HT in brain tissue.
5. An antidepressant product comprising lactobacillus paracasei E6 as claimed in claim 1.
6. An antidepressant product according to claim 5, wherein said product is a probiotic preparation.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018220416A1 (en) * | 2017-05-31 | 2018-12-06 | Compagnie Gervais Danone | Lactobacillus paracasei strain capable of improving the immune response to a viral-bacterial coinfection |
| CN111560331A (en) * | 2020-05-15 | 2020-08-21 | 诺佰克（武汉）生物科技有限公司 | Lactobacillus paracasei and application thereof |
| KR20220040149A (en) * | 2020-09-23 | 2022-03-30 | 주식회사 에치와이 | Compositions for Preventing, Improving or Treating Cognitive Disorder and Depression |
| CN114717148A (en) * | 2022-03-17 | 2022-07-08 | 江南大学 | Lactobacillus paracasei CCFM1229 for relieving depression mood and application thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018220416A1 (en) * | 2017-05-31 | 2018-12-06 | Compagnie Gervais Danone | Lactobacillus paracasei strain capable of improving the immune response to a viral-bacterial coinfection |
| CN111560331A (en) * | 2020-05-15 | 2020-08-21 | 诺佰克（武汉）生物科技有限公司 | Lactobacillus paracasei and application thereof |
| KR20220040149A (en) * | 2020-09-23 | 2022-03-30 | 주식회사 에치와이 | Compositions for Preventing, Improving or Treating Cognitive Disorder and Depression |
| CN114717148A (en) * | 2022-03-17 | 2022-07-08 | 江南大学 | Lactobacillus paracasei CCFM1229 for relieving depression mood and application thereof |
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