Application of Lactobacillusparacasei L.pR3-10 in preparation of drug for relieving anxiety and improving sleep

A technology to improve sleep and Lactobacillus, applied in the field of microorganisms, can solve the problems of lack of scientific research evidence on the function of probiotics, not necessarily suitable, and few probiotics

Active Publication Date: 2021-04-30
广东南芯医疗科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there are few studies and applications of probiotics in relieving anxiety and improving sleep
At the same time, the current international patent applications for probiotics are concentrated in the United States, Japan, and Russia, traditional research and development powerhouses, while my country lacks functional strains with independent intellectual prop

Method used

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  • Application of Lactobacillusparacasei L.pR3-10 in preparation of drug for relieving anxiety and improving sleep
  • Application of Lactobacillusparacasei L.pR3-10 in preparation of drug for relieving anxiety and improving sleep
  • Application of Lactobacillusparacasei L.pR3-10 in preparation of drug for relieving anxiety and improving sleep

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1L

[0024] The isolation and identification of the precursor strain L.p R3 (Lactobacillus paracasei) of embodiment 1L.p R3-10

[0025] (1) Sample source

[0026] Healthy babies aged 0-6 months were selected from the families of school staff as volunteers. Two weeks before sampling, a normal diet is required and there is no recent history of intestinal infection or antibiotic use. Take the morning stool on the day of sampling, and quickly use the intelligent microbial separation system of Nanjing Famet Company to separate the fecal bacteria after collection. After the separation was completed, the crude fecal bacteria liquid was collected quickly, added with cryopreservation protection solution, and then stored in a -80°C ultra-low temperature refrigerator for future use.

[0027] (2) Isolation, culture and identification of L.p R3

[0028] Take 1mL of the crude fecal extract liquid and add it to 9mL of normal saline, mix well and perform gradient dilution. Draw the dilution con...

Embodiment 2L

[0031] Induction and identification of embodiment 2L.p R3-10

[0032] (1) Low nutrient gradient tolerance method induces L.p R3 to become L.p R3-10

[0033]Take out the frozen L.p R3 from the -80°C refrigerator and put it in a warm water bath at 37°C to melt it quickly. The thawed bacterial solution was poured into an anaerobic blood agar plate, and placed in an anaerobic condition at 37°C for static culture for 48 hours. Observe the growth of the colony on the plate and whether there is a hemolytic ring, observe the shape of the strain under a Gram staining microscope, and confirm that there is no pollution, transfer to the MRS agar plate, culture it under anaerobic conditions at 37°C for 24 hours, and pick a single cell on the plate. The colonies were inoculated in 6mL MRS liquid medium, and cultured and activated under anaerobic conditions at 37°C for 16-18h. The activated bacterial solution was inoculated into 100 mL of MRS broth with an inoculum amount of 3% (v / v), cult...

Embodiment 3

[0043] Preparation of embodiment 3 Lactobacillus paracasei L.p R3-10 fermentation supernatant (extracellular secretion), bacterium suspension (thalline)

[0044] Lactobacillus paracasei L.p R3-10 was activated and cultured and inoculated in MRS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 1×10 7 CFU / mL, centrifuged at 4°C, 6000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 1×10 7 CFU / mL to obtain bacterial suspension (bacteria). Fermentation supernatant (extracellular secretions) and bacterial suspension (bacteria) were heated at 121°C for 20 minutes to prepare heat-inactivated fermentation supernatant (extracellular s...

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Abstract

The invention discloses application of Lactobacillusparacasei L.pR3-10 in preparation of a drug for relieving anxiety and improving sleep, and belongs to the technical field of microorganisms. The Lactobacillusparacasei L.pR3-10 inactivated and non-inactivated fermentation supernatant and bacterial suspension disclosed by the invention can obviously reduce the movement distance, mania time and active time of zebra fish and obviously prolong the standing time in an in-vivo insomnia model, and have the potential of being applied to relieving anxiety and improving sleep in vivo. The Lactobacillusparacasei L.pR3-10 disclosed by the invention has a huge potential application prospect in the aspects of relieving anxiety and improving sleep.

Description

technical field [0001] The invention relates to the technical field of microbes, and more specifically relates to the application of Lactobacillus paracasei L.p R3-10 in preparing medicines for relieving anxiety and improving sleep. Background technique [0002] Insomnia is a common disorder characterized by difficulty falling asleep, staying asleep, or waking up too early. Approximately 20-30% of adults have chronic insomnia problems (ie, lasting at least 1 month), and it is the most common sleep disorder in the general population. It is often accompanied by mental or physical disorders such as impaired concentration, restlessness, restlessness, and anxiety. Insomnia also increases the risk of chronic diseases such as high blood pressure, diabetes, stroke and coronary artery disease. Although medications for insomnia vary, most are not recommended for long-term use due to potential adverse effects. Therefore, there is a need to find alternative methods of treating insomn...

Claims

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Application Information

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IPC IPC(8): A61K35/747A61P25/20A61P25/22C12N1/20C12R1/225
CPCA61K35/747A61P25/20A61P25/22C12N1/20Y02A50/30
Inventor 张召郑康帝马燕曾今成陈涛
Owner 广东南芯医疗科技有限公司
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