Application of microorganism containing 3 beta-steroid dehydrogenase gene as marker of male depression

A technology of steroid dehydrogenase and depression, which is applied in the field of medicine, can solve problems such as depression testosterone deficiency, primary secondary testosterone reduction, and not fully explained

Pending Publication Date: 2022-01-21
李艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In some cases, there will be primary (such as Klinefelter's syndrome and mumps orchitis, etc.) or secondary (such as pituitary damage and excessive starvation) low testosterone, but the current cause of testosterone deficiency in depression is largely not fully explained

Method used

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  • Application of microorganism containing 3 beta-steroid dehydrogenase gene as marker of male depression
  • Application of microorganism containing 3 beta-steroid dehydrogenase gene as marker of male depression
  • Application of microorganism containing 3 beta-steroid dehydrogenase gene as marker of male depression

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Experimental program
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Effect test

Embodiment 1

[0048] Based on the hypothesis that the degradation of testosterone by bacteria in the intestinal tract can reduce serum testosterone, the inventors mixed 0.5 g of fresh stool samples from patients with depression with 50 mL of selective medium (2mM MgSO 4 ·7H 2 O, 0.1mM CaCl 2 2H 2 O, 48mM Na 2 HPO 4 , 22mM KH 2 PO 4 , 9mM NaCl, 19mM NH 4 Cl and 3mg / L testosterone) mixed. Under anaerobic or aerobic culture conditions, culture on a rotary shaker at 37°C and 250 rpm in the dark. When the concentration of testosterone in the medium was observed to decrease to 40%, 1 mL of the culture suspension was transferred to a 250 mL Erlenmeyer flask containing 49 mL of fresh selection medium. This process was repeated five times over a 26-day period, and the culture suspension from the fifth Erlenmeyer flask was inoculated onto Columbia blood agar plates to isolate testosterone-degrading strains such as figure 1 shown.

[0049] The strain was identified as Mycobacterium neoaurum ...

Embodiment 2

[0052] In order to explore whether M.neoaurum in the intestine can induce depression-like behavior in rats, we administered M.neoaurum to rats by intragastric administration. Animal studies were performed in strict accordance with the Experimental Animal Care and Use Protocol (S0271902190A) approved by the Animal Experiment Center of Renmin Hospital of Wuhan University. SD rats (male, 6 weeks old, 6-12 per group) were purchased from Hunan SJA Experimental Animal Company (Hunan, China), and used for the experiment after being fed and adapted to Specific Pathogen Free (SPF) in the Animal Experiment Center of Renmin Hospital of Wuhan University for one week . Rats were housed under SPF conditions on a 12-h light and dark schedule in a humidity-controlled and temperature-controlled (22°C-25°C) environment with autoclaved food and autoclaved water.

[0053] The two-way interaction between gut microbes and the brain is known as the “Microbiota–Gut–Brain” (MGB) axis. The MGB axis i...

Embodiment 3

[0058] We speculate that the depressive-like behavior of M. neoaurum is due to its ability to degrade testosterone. Therefore, if a testosterone-degrading bacterium is constructed through genetic engineering, then the bacterium can also cause depression-like behavior in rats by gavage. In order to construct recombinant Escherichia coli, we tried to find the gene sequence of testosterone degrading enzyme in M.neoaurum. By performing whole-genome complete map sequencing on the strain of M.neoaurum we isolated, we obtained the whole-genome sequence information. In our whole genome of M.neoaurum chromosome (66.26% GC, accession number: CP074376), the gene MN2019_09885 encoding 3β-steroid dehydrogenase (3β-HSD) was locked by us.

[0059] In order to construct testosterone-degrading bacteria and verify that the protein expressed by this sequence has testosterone-degrading ability, we performed heterologous expression. Plasmid pET28a was used as an overexpression vector to increase...

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Abstract

The invention relates to application of a microorganism containing a 3 beta-steroid dehydrogenase gene as a marker of male depression. Intestinal bacteria capable of degrading testosterone are successfully separated from a feces sample of a depression patient, and are identified as mycobacterium neoaureum through sequencing. Furthermore, a behavior experiment proves that the mycobacterium neoaureum can reduce serum testosterone and induce a depression-like behavior of a rat, and the depression-like behavior of the rat is reduced after targeted antibiotic treatment. Furthermore, whole genome sequence information is obtained by performing whole genome completion graph sequencing on the mycobacterium neoaureum, and the gene for encoding 3 beta-steroid dehydrogenase is locked, thus that 3 beta-steroid dehydrogenase can degrade testosterone into androstenedione to cause the depression-like behavior is proved. Therefore, the related microorganism can be used as the marker of male depression, and a corresponding detection agent can be used for evaluating the risk of a male subject suffering from depression or judging the pathogenesis of the male depression patient.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of a microorganism containing 3β-steroid dehydrogenase gene as a marker of male depression. Background technique [0002] Depressive disorders are a global public health problem. In 2017, the World Health Organization announced that depression will become the leading cause of disability worldwide. The incidence of depression in men increases with the onset of age-related testosterone decline, but men's testosterone does not decline in old age. Testosterone deficiency occurs in approximately 6% of the US male general population and in approximately 39% of the male outpatient population. In some cases, there will be primary (such as Klinefelter's syndrome and mumps orchitis, etc.) or secondary (such as pituitary damage, excessive starvation, etc.) testosterone reduction, but the current cause of testosterone deficiency in depression is largely not fully explaine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/04A61K45/00A61P25/24C12R1/32
CPCC12Q1/689C12Q1/04A61K45/00A61P25/24G01N2333/35G01N2800/304
Inventor 李艳李迪
Owner 李艳
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