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Application of Apelin-13 in preparation of drugs for treating cerebral trauma

A brain trauma and drug technology, applied in the field of biomedicine, can solve the problem that the mechanism of action is not completely clear.

Pending Publication Date: 2018-11-13
包海军
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2007, Simpkin et al. confirmed that Apelin-13 played a very good protective role in both in vivo and isolated experimental models of myocardial ischemia-reperfusion, but so far, most studies have been limited to its role in the cardiovascular system and significance, its specific mechanism of action is not fully understood

Method used

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  • Application of Apelin-13 in preparation of drugs for treating cerebral trauma
  • Application of Apelin-13 in preparation of drugs for treating cerebral trauma
  • Application of Apelin-13 in preparation of drugs for treating cerebral trauma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Cell Culture Reagent Preparation

[0020] 1.1 High glucose DMEM cell culture medium

[0021] High glucose DMEM cell culture medium (powder) 13.4g, 3.7g NaHCO 3 1. Double antibodies (100 U / mL penicillin and 100 μg / mL streptomycin) were dissolved in 1 L of double distilled water, stirred with a magnetic stirrer until completely dissolved, and the pH was adjusted to 7.1-7.2. Sterilize by positive pressure filtration with a 0.22μm pore size filter membrane, pack in sterilized 100mL glass bottles, and store in a refrigerator at 20°C. Samples were taken and cultured in a constant temperature incubator at 37°C for sterility test.

[0022] 1.2 RPMI 1640 cell culture medium

[0023] Take a pack of RPMI 1640 medium (powder) and 2g NaHCO 3 Dissolve in 1L double-distilled water, and the remaining steps are the same as the configuration of high-glucose DMEM medium.

[0024] 1.3 Trypsin working solution (0.25%)

[0025] 2.5g of trypsin and 0.2g of EDTA were dissolved ...

Embodiment 2

[0028] Embodiment 2 reagent preparation

[0029] 2.1 Drug preparation

[0030] 1) NGF stock solution: Add 1 mL of sterile PBS containing 0.1% bovine serum albumin to the freeze-dried powder to make a 100 μg / mL stock solution, divide into 10 tubes, 100 μL per tube, and store at -80 °C. Each tube was further diluted with sterile PBS to a final concentration of 10 μg / mL for a total of 1 mL. When used, dilute it with medium to make the working solution concentration 50ng / mL.

[0031] 2) Thrombin stock solution: Add 10 mL of sterile PBS containing 0.1% bovine serum albumin to each bottle of lyophilized powder, mix well and filter to make the stock solution concentration 100 U / mL, store at -20°C. Dilute it to the working solution concentration of 25U / mL (PC-12 cells) and 75U / mL (HT-22 cells) before use.

[0032] 3) Apelin-13 stock solution: take 5 mg of Apelin-13 and add 1 mL of PBS to prepare 5 μg / μL. According to its molecular weight is 1550.8, calculate the molar concentratio...

Embodiment 3

[0057] Embodiment 3 experimental method

[0058] 3.1 Cell Culture

[0059] Mouse hippocampal neuron HT-22 cells in RPIM-1640 medium, 10% FBS (Hangzhou Sijiqing), 37°C, 5% CO 2 cultivated under conditions.

[0060] Rat adrenal pheochromocytoma PC-12 cells in high-glucose DMEM medium, 10% FBS (Hangzhou Sijiqing), 37°C, 5% CO 2 Under the conditions of culture, 50ng / mL NGF needs to be added to induce the differentiation of cells into neurons.

[0061] 3.1.1 Cell Recovery

[0062] 1) Take out the cryopreservation tube containing HT-22 or PC-12 cells from the liquid nitrogen, put it into a 37°C water bath quickly, and let it melt (about 1min).

[0063] 2) Dilute the frozen cell solution to 3ml with RPIM-1640 or DMEM basal medium at room temperature.

[0064] 3) Centrifuge at a speed of 1000 rpm for 5 minutes.

[0065] 4) Discard the supernatant, resuspend the cells with RPIM-1640 or DMEM complete medium, transfer to the cell culture dish, 37°C, 5% CO 2 nourish.

[0066] 3.1....

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Abstract

Provided is application of Apelin-13 in preparation of drugs for treating cerebral trauma. Proliferation of nerve cells after brain injuries is promoted and the level of mitochondrial autophagy of nerve cells after thrombin injuries in the PTEN / Akt / FoxO3a / BNIP3 signaling paths is inhibited through Apelin-13, and a new way is provided for preparation of drugs for repairing nerve cells after brain injuries and drugs for treating brain injuries.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of Apelin-13 in the preparation of medicines for treating brain trauma. Background technique [0002] In 1998, Tatemoto et al. extracted and purified the natural ligand of APJ from the secretion of cow stomach by the same method as the discovery of the prolactin-releasing peptide hGR3, and named it Apelin, and he also found that Apelin-13, Apelin-36 Among the three subtypes of Apelin-17 and Apelin-17, Apelin-13 has the highest receptor affinity. Due to the wide distribution of Apelin in the body, it plays an important role in cardiovascular, immune, neuroendocrine and many other aspects. Apelin-13 has a very good effect on protecting the brain from focal ischemia-reperfusion injury, and can significantly reduce its cerebral edema and cerebral ischemia area, but its specific mechanism is unknown. In 2007, Simpkin et al. confirmed that Apelin-13 played a very good prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/10A61P25/00
CPCA61K38/10A61P25/00
Inventor 包海军蒯锦霞李周儒董国凯马静媛殷文江孙晓明
Owner 包海军
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