Application of Lactobacillus paracasei l.p R3-10 in the preparation of anxiety-relieving and sleep-improving drugs

A technology for improving sleep and Lactobacillus, applied in the field of microorganisms, can solve problems such as not necessarily suitable, few probiotics, and dependence on imports of probiotic strains, and achieve the effects of increasing resting time, reducing exercise distance, and relieving anxiety

Active Publication Date: 2022-07-08
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 property rights
The probiotic strains used by domestic production enterprises have long relied on imports, and foreign strains may not be suitable for the physiological conditions of the gastrointestinal tract of Chinese residents
In addition, the function of probiotics lacks strong scientific research evidence, which seriously affects the promotion of probiotics and their products

Method used

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  • Application of Lactobacillus paracasei l.p R3-10 in the preparation of anxiety-relieving and sleep-improving drugs
  • Application of Lactobacillus paracasei l.p R3-10 in the preparation of anxiety-relieving and sleep-improving drugs
  • Application of Lactobacillus paracasei l.p R3-10 in the preparation of anxiety-relieving and sleep-improving drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1L

[0024] Example 1 Isolation, culture and identification of the precursor strain L.p R3 (Lactobacillus paracasei) of L.p R3-10

[0025] (1) Sample source

[0026] Healthy infants aged 0-6 months were selected from school staff families as volunteers. Two weeks before sampling, normal diet was required and there was no recent history of intestinal infection and antibiotic use. The morning stool was taken on the day of sampling. After collection, the intelligent microbial separation system of Nanjing Famet Company was used to separate fecal bacteria. After the separation was completed, the crude fecal bacterial liquid was collected quickly, added with cryopreservation protection solution, and then placed 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 crude fecal bacteria solution and add it to 9mL of normal saline, mix well and then carry out gradient dilution. Aspirate dilution con...

Embodiment 2L

[0031] Example 2 Induction and identification of L.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 freezer, and put it in a 37°C warm water bath to thaw it quickly. The thawed bacterial liquid was poured into an anaerobic blood agar plate, and placed under anaerobic conditions at 37 °C for 48 h. Observe the growth of colonies in the plate and the formation of hemolytic rings. Observe the morphology of the strains under a Gram stain microscope. After confirming that there is no contamination, transfer them to MRS agar plates and place them in anaerobic conditions at 37 °C for 24 hours. Pick a single colony on the plate. Colonies were inoculated into 6 mL of MRS liquid medium, and cultured under anaerobic conditions at 37 °C for 16-18 h. The activated bacterial solution was inoculated into 100 mL of MRS broth with a 3% (v / v) inoculum, 120 rpm / min, and shaken at 37° C. for 16-18 h. ...

Embodiment 3

[0043] Example 3 Preparation of Lactobacillus paracasei L.p R3-10 fermentation supernatant (extracellular secretion), bacterial suspension (cell)

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

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Abstract

The invention discloses the application of Lactobacillus paracasei L.pR3-10 in the preparation of a drug for relieving anxiety and improving sleep, and belongs to the technical field of microorganisms. The inactivated and non-inactivated fermentation supernatants and bacterial suspensions of Lactobacillus paracasei L.pR3-10 disclosed in the invention can significantly reduce the movement distance, manic time and active time of zebrafish in the in vivo model of insomnia. And significantly increase resting time, has the potential to be used in vivo to relieve anxiety and improve sleep. A strain of Lactobacillus paracasei L.pR3-10 disclosed in the invention has a huge potential application prospect in relieving anxiety and improving sleep.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to the application of Lactobacillus paracasei L.p R3-10 in the preparation of medicines for relieving anxiety and improving sleep. Background technique [0002] Insomnia is a common disorder characterized by difficulty falling asleep, difficulty maintaining sleep, or waking up too early. About 20-30% of adults have chronic insomnia problems (ie, lasting at least 1 month), which is the most common sleep disorder in the general population. It is often accompanied by mental or physical disorders such as diminished concentration, restlessness, restlessness, 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 inso...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04A61K35/747A61P25/20A61P25/22C12N1/20C12R1/225
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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