Application of brown adipocyte product in preparation of medicine for preventing and treating osteoporosis

A technology for brown fat and osteoporosis, applied in drug combinations, bone diseases, pharmaceutical formulations, etc., can solve problems such as atypical femoral fractures and osteonecrosis of the jaw, increase patients' BMD levels, and produce nausea, and achieve good implementability , increase bone volume fraction and thickness, and promote osteogenic differentiation

Active Publication Date: 2021-06-01
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can increase the BMD level of patients and reduce the risk of vertebral, non-vertebral and hip fractures, but may have adverse reactions of skin infection
Teriparatide is a synthetic 1-34 amino acid fragment of human parathyroid hormone (PTH), which can increase BMD levels, but may cause adverse reactions such as nausea, headache and dizziness
Romosozumab is a humanized antibody that binds sclerostin with high affinity and increases bone mineral density, but long-term drug treatment can cause atypical femoral fractures in the subtrochanteric region and femoral shaft and jaw fractures Adverse reactions such as necrosis

Method used

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  • Application of brown adipocyte product in preparation of medicine for preventing and treating osteoporosis
  • Application of brown adipocyte product in preparation of medicine for preventing and treating osteoporosis
  • Application of brown adipocyte product in preparation of medicine for preventing and treating osteoporosis

Examples

Experimental program
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Effect test

Embodiment 1

[0047] The preparation of embodiment 1 brown adipocyte product

[0048] 1. Take 5 mice for the following experimental process: euthanize 4-week-old C57BL / 6j mice, thoroughly disinfect them with 70% alcohol, dissect out the butterfly-shaped brown adipose tissue in the interscapular region of the mice in an ultra-clean bench, and put Put it into a culture dish containing PBS, wash away the impurities on the surface of the brown adipose tissue and remove the surrounding excess tissue, transfer the tissue into a new culture dish and cut it into the smallest visible tissue pieces with scissors, then transfer to a 50ml centrifuge tube and add 1mg / ml Type 1 collagenase and DMEM with 1% bovine serum albumin were placed in a 37°C water bath and shaken for 30 minutes, then neutralized the digestive enzymes with DMEM containing 10% bovine serum albumin, filtered through a 70μm filter into a new 50ml centrifuge tube, Centrifuge at 300g for 5 minutes to obtain brown adipose precursor cells...

Embodiment 2

[0054] Example 2 Effect of Brown Adipocyte Products on Bone Marrow Mesenchymal Stem Cells

[0055] The brown adipocyte product (BAT CM) was co-cultured with bone marrow mesenchymal stem cells (BMSC), and osteogenic induction medium was used to induce osteogenesis, and alkaline phosphatase staining and RNA extraction were performed on day 14 to verify the brown adipocyte product Alizarin red staining was performed 21 days after induction of differentiation for the effect on BMSC osteogenic differentiation, and semi-quantitative detection was performed. image 3 Middle A is the results of alkaline phosphatase (ALP) and Alizarin Red (AlizarinRed) staining on days 14 and 21 after co-culture of brown adipocyte products and BMSCs, respectively. image 3 Middle B shows that BAT CM promotes mineralized nodule formation of BMSCs by semi-quantitative alizarin red staining. image 3 Middle C shows RT-PCR verification of BMSCs co-cultured for 14 days, and it was found that genes related ...

Embodiment 3

[0056] Example 3 Effects of brown adipocyte products on bone marrow mononuclear macrophages

[0057] Brown adipocyte products (BAT CM) were co-cultured with bone marrow mononuclear macrophages (BMM), and 30ng / ml of M-csf and 50ng / ml of RANKL were added to induce osteoclast differentiation, and 6 days later, anti-tartaric acid (TRAP ) staining, osteoclast count and RT-PCR to verify the effect of BAT CM on osteoclast differentiation. Figure 4 Middle A shows the results of TRAP staining after 6 days of induction of BMM differentiation, showing fewer osteoclasts in the BAT CM group. Figure 4 middle B pair Figure 4 The number of osteoclasts in A was counted, showing that the number of osteoclasts in the BAT CM group was less. Figure 4 Middle C shows that 6 days after BMM induction and differentiation, RT-CPR was performed to detect the expression levels of genes related to osteoclast differentiation, which showed that the gene expression levels in the BAT CM group were lower ...

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Abstract

The invention discloses a preparation method of a brown adipocyte product, and the brown adipocyte product obtained by adopting the corresponding preparation method can promote osteogenic differentiation of BMSC, reduce the number of osteoclasts, reduce the gene expression level related to osteoclast differentiation, increase the volume fraction and thickness of cortical bones and cancellous bones. The product can be used for preparing medicines for preventing and treating osteoporosis.

Description

technical field [0001] The invention belongs to the technical field of new uses of medicines, and in particular relates to the application of brown adipocyte products in the preparation of medicines for preventing and treating osteoporosis. Background technique [0002] Adipose tissue in the body is divided into white adipose tissue (WAT), brown adipose tissue (BAT) and beige adipose tissue (beige adipose tissue, BAT). The main function of WAT is to store energy; BAT is to produce Hot adipose tissue mainly exists in the interscapular area of ​​newborns, which can increase the energy metabolism of the body to maintain the body temperature of newborns through non-shuddering heat production; beige adipose tissue is scattered in white adipose tissue and has a function similar to brown fat Thermogenic fat, exercise, cold, and beta 3 adrenoceptor agonists all promote beige fat production [1,2]. Brown adipocytes and muscle fiber cells are derived from Pax7+ / Myf5+ stem cells, while...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/35A61P19/10
CPCA61K35/35A61P19/10Y02A50/30
Inventor 于志锋杜静珂
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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