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: 2020-07-10
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 t

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: Effect of 2-APB on the Viability of SNP-Induced Primary Articular Chondrocytes

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

[0024] (1) Take a number of male SD rats weighing about 160g SPF grade, put them to death by bleeding from the femoral artery after anesthesia, immerse them in 0.1% bromogeramine for sterilization, cut off the hind legs of the knee joints by the outer flame of the alcohol lamp, peel off the fur, and place Rinse with normal saline and move to ultra-clean bench for operation;

[0025] (2) Disinfect the surgical instruments in the ultra-clean table before using them, use tweezers to clamp out the hind limbs, cut the knee joint crosswise with a scalpel, and gently peel off the muscle ligaments at the joint to completely expose the articular cartilage, then scrape gently Hyaline cartilage slices are placed in a small plate containing PBS (double antibody);

[0026] (3) Gently rinse the articular cart...

Embodiment 2

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

[0043] Experimental method: the extraction, separation and cultivation of the primary rat articular chondrocytes were the same as in Example 1, and the primary rat articular chondrocytes were mixed at 2×10 per well. 5After inoculation at a density of 300 cells, an in vitro model of apoptosis was established by adding the apoptosis inducer SNP (0.5 mM). In vitro experiments, the cells were divided into normal group, SNP (0.5mM) group, SNP (0.5mM) + different concentrations of 2-APB (50, 100, 200uM) combined drug group, the corresponding drugs were treated with chondrocytes for 12 hours, and lactate dehydrogenase was measured after the experiment ( LDH) release. Detection method of LDH release:

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

[0045] (2) Observe under a microscope, when the cells in each w...

Embodiment 3

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

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

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Abstract

The invention relates to application of 2-aminoethoxy diphenyl borate in preparation of a medicine for treating osteoarthritis, and belongs to the technical field of biological medicines. According tothe invention, an apoptosis model is constructed on primary rat articular cartilage cells to simulate cartilage cell damage after osteoarthritis. The invention finds that 2-APB with different concentrations can obviously recover the cell viability and mitochondrial membrane potential level of apoptotic chondrocytes and reduce LDH leakage rate and ROS release, and can reduce the cartilage cell apoptosis by inhibiting IHH/Bcl-2/Bax signal transduction pathway, such that the related research on the cartilage cell protection mechanism is further supplemented and enriched, and the more scientificbasis is provided for the research and the development of new drugs for treating osteoarthritis.

Description

technical field [0001] The invention relates to the application of 2-aminoethoxydiphenyl borate (2-APB) in the preparation of medicines for treating osteoarthritis, and belongs to the technical field of biomedicine. Background technique [0002] Osteoarthritis (OA) is the result of the combined effects of obesity, strain, aging, trauma and other causes. 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, osteosclerosis, cystic degeneration, osteophytes, and joint hypertrophy and deformation constitute the pathological core of osteoarthritis, especially articular cartilage degeneration as the main lesion, and chondrocyte apoptosis as the main change at the microscopic level. At present, the drugs for the treatment of OA are mainly anti-inflammatory and analgesic, but the failure to inhibit the destru...

Claims

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Application Information

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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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