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Application of 2-apb in preparation of medicine for treating osteoarthritis

A 2-APB, 1.2-APB technology, applied in the field of biomedicine, can solve the problem of no 2-APB, and achieve the effects of reducing chondrocyte apoptosis, reducing LDH leakage rate, and restoring cell vitality

Active Publication Date: 2022-05-24
THE SECOND AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no report on the preparation of 2-APB for the preparation of drugs for the treatment of osteoarthritis

Method used

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  • Application of 2-apb in preparation of medicine for treating osteoarthritis
  • Application of 2-apb in preparation of medicine for treating osteoarthritis
  • Application of 2-apb in preparation of medicine for treating osteoarthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: The effect of 2-APB on the viability of primary articular chondrocytes induced by SNP

[0023] 1. Extraction, isolation and culture of primary rat articular chondrocytes

[0024] (1) Take several male SD rats with a weight of about 160g of SPF grade, femoral artery bloodletting after anesthesia, sterilization by immersion in 0.1% Syngeeridine, cut off the hind legs of the knee joint beside the outer flame of an alcohol lamp, peel off the fur, and put them in Rinse in normal saline and move to a clean bench for operation;

[0025] (2) Disinfect the surgical instruments in the ultra-clean bench and use them. Use tweezers to clip out the hind limbs. Cut the knee joint with a scalpel. Gently peel off the muscles and ligaments at the joint to completely expose the articular cartilage. Gently scrape The hyaline cartilage slices were placed in a small plate containing PBS (double antibody);

[0026] (3) Rinse the articular cartilage gently with PBS (double antibod...

Embodiment 2

[0042] Example 2: Effect of 2-APB on SNP-induced LDH release from primary articular chondrocytes

[0043] Experimental method: The extraction, isolation and culture of primary rat articular chondrocytes were the same as those in Example 1. 5After seeding at a density of one cell, an in vitro model of apoptosis was established by adding the apoptosis-inducing agent SNP (0.5 mM). In vitro experiment cells were divided into normal group, SNP (0.5mM) group, SNP (0.5mM) + different concentrations of 2-APB (50, 100, 200uM) combined treatment group, the chondrocytes were treated with corresponding drugs for 12h, and lactate dehydrogenase (LDH) was measured after the experiment. LDH) release amount. Detection method of LDH release:

[0044] (1) Digest the cells in the culture flask and inoculate them in a 24-well plate with 400ul (1×10 4 cells);

[0045] (2) Observe under the microscope, when the cells in each well grow to 60%-70%, starve for 12 hours, and then give corresponding ...

Embodiment 3

[0052] Example 3: Effect of 2-APB on SNP-induced apoptosis of primary articular chondrocytes

[0053] Experimental method: After sterilizing the cover glass with alcohol, put it into a 6-well culture plate, then inoculate primary articular chondrocytes in the 6-well culture plate, transfer it to a cell incubator for culture, and wait until the cells cover 70% of the culture plate. -80%, divided into normal group (Control), SNP (0.5mM) group, 2-APB (100uM) group and SNP (0.5mM) + 2-APB (100uM) combination group, the chondrocytes were treated with corresponding drugs for 12h , perform Hochst 33342 staining operation: take the 6-well plate out of the incubator and put it into the ultra-clean bench, discard the liquid, wash 2 times with PBS, discard the PBS, take out the coverslip, and fix the cells with 4% paraformaldehyde for about 15 minutes , washed 3 times with PBS, discarded PBS, added 25ug / ml of Hochst 33342 fluorescent dye in the dark, incubated at 37 °C for 15 min in the ...

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Abstract

The invention relates to the application of 2-aminoethoxy diphenyl borate in the preparation of medicines for treating osteoarthritis, and belongs to the technical field of biomedicine. The present invention simulates chondrocyte damage after osteoarthritis by constructing an apoptosis model on primary rat articular chondrocytes, and finds that different concentrations of 2-APB can significantly restore the cell viability and mitochondrial membrane potential level of apoptotic chondrocytes, reduce LDH leakage rate, ROS release, and can reduce chondrocyte apoptosis by inhibiting the IHH / Bcl‑2 / Bax signal transduction pathway, which further complements and enriches the research on the protection mechanism of chondrocytes, and provides a basis for the research and development of osteoarthritis The new drug provides more scientific basis.

Description

technical field [0001] The invention relates to the application of 2-aminoethoxydiphenyl borate (2-APB) in preparing a medicine for treating osteoarthritis, and belongs to the technical field of biomedicine. Background technique [0002] Osteoarthritis (OA) is the result of the joint action of various causes such as obesity, strain, aging, and trauma. It is a degenerative inflammatory joint disease. The main pathological changes of OA are joint swelling and pain and different degrees of dysfunction. , leading to loss of function in severe cases. Cartilage degeneration and wear, bone sclerosis, cystic degeneration, osteophytes, and joint hypertrophy and deformation constitute the pathological core of osteoarthritis. In particular, articular cartilage degeneration is the main lesion, and microscopically, chondrocyte apoptosis is the main change. The current drugs for the treatment of OA are mainly anti-inflammatory and analgesic, but the failure to inhibit the destruction of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/69A61P19/02A61P29/00
CPCA61K31/69A61P19/02A61P29/00
Inventor 周仁鹏胡伟陈勇马刚刚鲁超
Owner THE SECOND AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
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