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CN-121971495-A - Application of bifidobacterium animalis subspecies lactis in improving osteoporosis

CN121971495ACN 121971495 ACN121971495 ACN 121971495ACN-121971495-A

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to application of bifidobacterium animalis subspecies lactis in improving osteoporosis. The bifidobacterium animalis subspecies BL-136 provided by the invention is preserved in the microorganism strain collection in Guangdong province, the preservation number is GDMCC No:67831, and the strain has excellent osteoclast differentiation inhibiting capability and has obvious application advantages in the aspect of alleviating osteoporosis. The microgel preparation prepared by the strain can play an excellent role in relieving osteoporosis, and remarkably improves the curative effect of the osteoporosis. Therefore, the invention proves the curative effect of the bifidobacterium animalis subspecies BL-136 and the microgel preparation thereof on relieving osteoporosis, and can provide scientific thinking and technical support for the drug development of osteoporosis.

Inventors

  • LI BAILIANG
  • LV WENQING
  • Zhan Jingqi
  • FAN CHONGHUA

Assignees

  • 东北农业大学

Dates

Publication Date
20260505
Application Date
20260408

Claims (9)

  1. 1. A method for preparing a microgel preparation for improving osteoporosis, which is characterized by comprising the following steps: (1) Oxidizing konjak glucomannan to obtain oxidized konjak glucomannan, and performing sulfhydrylation modification on the oxidized konjak glucomannan to obtain sulfhydrylation oxidized konjak glucomannan; dissolving the sulfhydrylation oxidized konjak glucomannan and silk fibroin in water to obtain binary mixed solution; Preparing a heavy suspension of bifidobacterium animalis subspecies lactis, and centrifuging the heavy suspension to obtain bacterial mud; Re-suspending the bacterial sludge by adopting the binary mixed solution to obtain ternary mixed solution; (2) Preparing the ternary mixed solution into microdroplets, injecting the microdroplets into a ferrous ion solution for primary crosslinking, and injecting gel obtained by the primary crosslinking into a tannic acid solution for secondary crosslinking to obtain the microgel preparation for improving osteoporosis; the name of the bifidobacterium animalis subspecies lactis is bifidobacterium animalis subspecies (Bifidobacterium animalis subsp. Lactis) BL-136, and the bifidobacterium animalis subspecies are deposited in the Guangdong province microorganism strain collection, the deposition date is 2026, 02, 09 and the deposition number is GDMCC No:67831.
  2. 2. The method for preparing the microgel preparation for improving osteoporosis according to claim 1, wherein in the step (1), the oxidation treatment comprises the steps of dissolving konjak glucomannan in water, adding 2, 6-tetramethylpiperidine oxide and sodium bromide for mixing, adding sodium periodate for mixing, adjusting the pH value of a system to 10-13, and carrying out oxidation reaction for 3-5 hours to obtain oxidized konjak glucomannan, wherein in the oxidation treatment, the mass ratio of konjak glucomannan, 2, 6-tetramethylpiperidine oxide and sodium bromide is (1.8-2.2) to (0.05-0.08) to (0.2-0.4).
  3. 3. The method for preparing the microgel preparation for improving osteoporosis according to claim 1, wherein in the step (1), the sulfhydrylation modification comprises the steps of dissolving oxidized konjak glucomannan in water, then adding 1-ethyl (3-dimethylaminopropyl) carbodiimide hydrochloride for activation treatment for 0.5-2 h, and then adding L-cysteine hydrochloride monohydrate for reaction for 20-30 h, wherein the mass ratio of oxidized konjak glucomannan, 1-ethyl (3-dimethylaminopropyl) carbodiimide hydrochloride and L-cysteine hydrochloride monohydrate is (80-120) to (160-170) to (70-80) during sulfhydrylation modification.
  4. 4. The method for preparing a microgel preparation for improving osteoporosis according to claim 1, wherein in the step (1), the concentration of the thiol-modified konjac glucomannan in the binary mixture is 1.3% -1.7% w/v, and the concentration of the silk fibroin is 1.8% -2.2% w/v.
  5. 5. The method for preparing a microgel preparation for improving osteoporosis according to claim 1, wherein in the step (1), the concentration of viable bacteria in the heavy suspension of the bifidobacterium animalis subspecies is (0.5-2) x 10 9 CFU/mL, and the volume ratio of the heavy suspension of the bifidobacterium animalis subspecies to the binary mixed solution is (0.8-1.2) to 1.
  6. 6. The method for preparing the microgel preparation for improving osteoporosis according to any one of claims 1 to 5, wherein in the step (2), the preparation of the ternary mixed solution into droplets is performed by adopting an electrostatic spraying process, wherein the electrostatic spraying process is carried out under the conditions that the applied voltage is 8k to 16kV, the distance from the tip to the current collector is 8 cm to 15cm, and the feeding rate is 0.5 ml/h to 0.8ml/h.
  7. 7. The method for preparing a microgel preparation for improving osteoporosis according to any one of claims 1 to 5, wherein in the step (2), the ferrous ion solution is a ferrous sulfate heptahydrate solution, the concentration of the ferrous sulfate heptahydrate solution is 0.5% -1.5% w/v, the primary crosslinking time is 1-3 hours, the concentration of the tannic acid solution is 0.5% -1.5% w/v, and the secondary crosslinking time is 1-3 hours.
  8. 8. A microgel formulation for improving osteoporosis prepared by the preparation method according to any one of claims 1 to 7.
  9. 9. Use of a microgel formulation according to claim 8 in the manufacture of a medicament for the treatment of osteoporosis.

Description

Application of bifidobacterium animalis subspecies lactis in improving osteoporosis Technical Field The invention belongs to the technical field of microorganisms, and particularly relates to application of bifidobacterium animalis subspecies lactis in improving osteoporosis. Background Osteoporosis is a systemic bone metabolism disorder characterized by reduced bone density, deterioration of bone microstructure and increased bone fragility, and constitutes a serious threat to the health and survival of the elderly population. The current osteoporosis prevalence rate of the elderly population in China is about 15%, and the patient scale is still continuously rising, so that the osteoporosis prevalence rate is a global major public health problem. At present, although the bone density can be improved to a certain extent in the clinical medicine treatment of osteoporosis, adverse reactions such as digestive tract reactions, thrombus formation risks and the like are often accompanied, and the long-term use safety of the medicine is still questioned. Therefore, the development of a safe and effective novel intervention strategy has become an important research direction in the field of osteoporosis prevention and treatment. The role of intestinal flora in the regulation of bone metabolism has gradually been of great interest. About 100 trillion microorganisms are symbiotic in the human intestinal tract, and are mainly distributed at the tail end of the colon, mainly comprising bacteroides, firmicutes and actinomycetes. The intestinal flora is not only involved in energy metabolism and maintains intestinal barrier function, but also affects the metabolic state of the distal organs via the "gut-bone axis". Bifidobacteria have shown a regulatory function on growth and development in early stages of life, but fewer dominant strains have been reported to be able to alleviate osteoporosis. Therefore, how to screen a specific strain capable of effectively relieving osteoporosis and further enhance the intervention effect of the specific strain on the osteoporosis through an embedding technology has important research significance and application value. Disclosure of Invention In order to overcome the defects in the prior art, the invention aims to provide the application of the bifidobacterium animalis subspecies in improving osteoporosis. The microgel preparation constructed by using the bifidobacterium animalis subspecies lactis BL-136 has remarkable improvement effect on osteoporosis, and can provide a technical basis for the development of probiotic medicines for treating osteoporosis. In order to achieve the above object, according to a first aspect of the present invention, a technical scheme is as follows: a method for preparing a microgel preparation for improving osteoporosis, which comprises the following steps: (1) Oxidizing konjak glucomannan to obtain oxidized konjak glucomannan, and performing sulfhydrylation modification on the oxidized konjak glucomannan to obtain sulfhydrylation oxidized konjak glucomannan; dissolving the sulfhydrylation oxidized konjak glucomannan and silk fibroin in water to obtain binary mixed solution; preparing a heavy suspension of the bifidobacterium animalis subspecies lactis, and centrifuging the heavy suspension to obtain bacterial sludge; Re-suspending the bacterial sludge by adopting the binary mixed solution to obtain ternary mixed solution; (2) Preparing the ternary mixed solution into microdroplets, injecting the microdroplets into a ferrous ion solution for primary crosslinking, and injecting gel obtained by the primary crosslinking into a tannic acid solution for secondary crosslinking to obtain the microgel preparation for improving osteoporosis; the name of the bifidobacterium animalis subspecies lactis is bifidobacterium animalis subspecies (Bifidobacterium animalis subsp. Lactis) BL-136, and the bifidobacterium animalis subspecies are deposited in the Guangdong province microorganism strain collection, the deposition date is 2026, 02, 09 and the deposition number is GDMCC No:67831. In the step (1), the oxidation treatment comprises the steps of dissolving konjak glucomannan in water, adding 2, 6-tetramethyl piperidine oxide and sodium bromide for mixing, adding sodium periodate solution, adjusting the pH value of the system to 10-13, and carrying out oxidation reaction for 3-5 hours to obtain the oxidized konjak glucomannan. In the oxidation treatment, the mass ratio of konjak glucomannan to 2, 6-tetramethylpiperidine oxide to sodium bromide is (1.8-2.2): (0.05-0.08): (0.2-0.4), more preferably 2:0.06:0.3. In the step (1), the sulfhydrylation modification comprises the following steps of dissolving oxidized konjak glucomannan in water, then adding 1-ethyl (3-dimethylaminopropyl) carbodiimide hydrochloride for activation treatment for 0.5-2 h, and then adding L-cysteine hydrochloride monohydrate for reaction for 20-30 h to obtain the sulfhydrylation oxidized konjak