Application of annexin a2 in the preparation of drugs for regulating the action of pth

A technology of annexin and action, which is applied in the application field of annexin A2 in the preparation of drugs that regulate the action of PTH, and can solve the problems of ineffective reduction of fractures and unsatisfactory treatment effects, etc.

Inactive Publication Date: 2017-01-04
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Practice has shown that although it can significantly increase bone mass, the therapeutic effect is not ideal, and it cannot effectively reduce the occurrence of fractures

Method used

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  • Application of annexin a2 in the preparation of drugs for regulating the action of pth
  • Application of annexin a2 in the preparation of drugs for regulating the action of pth
  • Application of annexin a2 in the preparation of drugs for regulating the action of pth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Effects on Alkaline Phosphatase Activity of Rat Osteoblast UMR106

[0033] 1. Recovery and passage of rat osteogenic sarcoma UMR106 cells

[0034] Take out the frozen rat osteogenic sarcoma UMR106 cells from liquid nitrogen, put them into a water bath at about 38°C to thaw, shake continuously during the period, until just completely thawed, then transfer to 8ml of serum-free DMEM that has been preheated to 37°C In the medium, mix well, centrifuge at 1000rpm for 5min, discard the supernatant, add about 5ml of DMEM medium containing 10% newborn calf serum to the precipitate to suspend, transfer to a 100ml cell culture bottle, and store at 37°C for 5 %CO 2 Cultivated in a saturated humidity carbon dioxide incubator. The next day, observe under an inverted biological microscope and change the medium, digest and passage with 0.25% trypsin in time, until the number of cells reaches the number required for the experiment, and proceed to the next experiment.

[0035] Add dif...

Embodiment 2

[0049] Effect on proliferation activity of rat osteoblast UMR106 (MTT method)

[0050] 1. Resuscitate rat osteoblastic sarcoma UMR 106 cells by the same method as in Example 1.

[0051] 2. Inoculate UMR106 cells: Digest monolayer cultured cells with 0.25% trypsin, prepare single cell suspension with DMEM medium containing 10% fetal bovine serum, and use 10% per well 3 ~10 4 The cells were seeded in a 96-well culture plate with a volume of 200 μl per well. Move the culture plate into CO 2 in an incubator at 37°C, 5% CO 2 and cultured under saturated humidity conditions. According to the method of Example 1, different gradients of PTH and protein samples combined with annexin A2 were added to induce UMR106 cells and stimulate cell proliferation for 72 hours.

[0052] 3. Color development: Add 10 μl of MTT solution (trade name: thiazolium blue, 5 mg / ml) to each well, continue to incubate in a 37°C incubator for 4 hours, terminate the culture, and carefully discard the cultur...

Embodiment 3

[0059] Effects on the generation of mineralized nodules by rat osteoblast UMR106

[0060] 1. Resuscitate rat osteoblastic sarcoma UMR 106 cells by the same method as in Example 1.

[0061] 2. Osteogenic induction: preparation of osteoblast culture medium

[0062] Rat osteogenic sarcoma UMR106 cells in the logarithmic growth phase were collected by digestion with 0.25% trypsin, and the cell concentration was adjusted to 2×10 with DMEM medium. 5 pieces / ml.

[0063] The experimental grouping method is the same as in Example 1. Cells were inoculated in a φ90mm cell culture dish, and proteins of corresponding concentrations were added respectively. The negative control group was cultured in osteoblast culture medium without any protein; the blank group was cultured in DMEM containing only 5% newborn calf serum. base. Each well of the experimental group and the negative control group was supplemented with osteoblast culture medium to 10ml, and the blank group was supplemented wi...

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Abstract

Application of annexin A2 in the preparation of medicines for regulating the action of PTH. Depending on the concentration of parathyroid hormone (PTH), annexin A2 can promote or inhibit the activity of PTH on ALP (alkaline phosphatase). This effect can at least realize the two-way regulation of the osteogenesis and osteoclast function of PTH with A2, and exert the effect of treating diseases including osteoporosis. Loose treatments are more precisely targeted and adaptable.

Description

technical field [0001] The invention relates to the application of annexin A2 in the preparation of drugs for regulating the action of parathyroid hormone (PTH), and can further regulate the influence of PTH on the activity of alkaline phosphatase (ALP) bidirectionally through annexin A2. This bi-directional regulation can be used at least in drugs for the treatment of bone diseases including osteoporosis. Background technique [0002] Taking osteoporotic bone diseases as an example, primary osteoporosis refers to the decrease in the proportion of bone matrix and bone mineral, and changes in the microstructure of bone tissue with increasing age (or postmenopausal women). , changes in normal loading, increased bone fragility, leading to susceptibility to spontaneous fractures or fractures with minor injuries. The occurrence of osteoporosis is the result of an imbalance between the bone resorption of osteoclasts (OC) and the bone formation of osteoblasts (OB), which leads to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P19/10
Inventor 陈雨雪白丁孟玉坤彭晓东魏大鹏
Owner SICHUAN UNIV
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