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Polypeptide for inhibiting cell proliferation and inducing apoptosis, and preparation method and use thereof

A technology for inhibiting cell and apoptosis, which is applied in the field of polypeptides that inhibit cell proliferation and induce apoptosis and the preparation thereof, achieves wide application prospects, improves long-term efficacy and safety, and has obvious effects.

Active Publication Date: 2016-08-17
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although the stent-coated drugs used clinically can significantly reduce the incidence of vascular restenosis, the overall incidence of restenosis is still high. rate is higher

Method used

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  • Polypeptide for inhibiting cell proliferation and inducing apoptosis, and preparation method and use thereof
  • Polypeptide for inhibiting cell proliferation and inducing apoptosis, and preparation method and use thereof
  • Polypeptide for inhibiting cell proliferation and inducing apoptosis, and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0041] Experimental Example 1: Detection of smooth muscle cell apoptosis by flow cytometry

[0042] 1. Take the rat VSMCs of the 4th-9th generation, and count them accurately according to 5×10 5 Cells / well were seeded in a six-well plate, and 2ml of DMEM (dulbecco's modified eagle medium cell culture medium) high-glucose medium was added. Three replicate wells could be set up in each group, and the 2 Continue culturing for 24 hours in a cell culture incubator.

[0043] 2. Synchronize, discard the medium in each experimental well, wash with sterile PBS (phosphate buffer solution) twice, pay attention to the operation to be gentle, try to avoid blowing up the cells, and then use a pipette gun to respectively Add 2ml of serum-free DMEM medium to each experimental well. Add SPHSG, MRSP and NC (invalid control) to each experimental group at a concentration of 25 μmol / L, and only add serum-free medium to the blank group, mix well and place in a cell incubator to continue culturing...

experiment example 2

[0049] Experimental Example 2: Detection of Caspase3 Enzyme Activity

[0050] 1. Take the rat VSMCs of the 4th-9th generation, and count them accurately according to 1×10 6 Seed each cell / well in a 60mm cell culture dish, add 3ml DMEM high-glucose medium, set up 3 duplicate wells for each group, and incubate at 37°C, 5% CO 2 Continue culturing for 24 hours in a cell culture incubator.

[0051] 2. Synchronize, discard the medium in each experimental well, wash with sterile PBS twice, pay attention to the operation to be gentle, try to avoid blowing up the cells, and then add 3ml serum-free DMEM to each experimental well with a pipette gun Medium. SPHSG, MRSP and NC were added to each experimental group at a concentration of 25 μmol / L, and only serum-free medium was added to the blank group, mixed evenly and placed in a cell culture incubator for 8 hours.

[0052] 3. Collect the cells when the cells have not entered the late stage of apoptosis, and collect the cells completel...

experiment example 3

[0062] Experimental example 3: In situ terminal transferase labeling technique (TUNEL) detection of apoptosis rate

[0063] 1. For cell slides, soak the autoclaved glass slides with 10% poly-slothine, air-dry them under sterile conditions, transfer them to a six-well plate, and set aside.

[0064] 2. Take the rat VSMCs of the 4th-9th generation, and count them accurately according to 2×10 5 Cells / well were seeded in a six-well plate, and 2ml DMEM high-glucose medium was added, and three replicate wells could be set up for each group, and the 2 Continue culturing for 24 hours in a cell culture incubator.

[0065] 3. Synchronize, suck up and discard the medium in each experimental well, wash with sterile PBS twice, pay attention to the operation to be gentle, try to avoid blowing up the cells, and then add 2ml serum-free DMEM to each experimental well with a pipette gun Medium. SPHSG, MRSP and NC were added to each experimental group at a concentration of 25 μmol / L, and only ...

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Abstract

The present invention discloses a polypeptide that inhibits cell proliferation and induces its apoptosis and its preparation method and application. The polypeptide of the present invention includes the polypeptide sequence "SPHSG" as: YGRKKRRQRRR‑GYLSKVRGISEVL; or is similar in homology and function to the polypeptide sequence The same another polypeptide sequence "MRSP" is: YGRKKRRQRRR‑DVKGYLSKVRGISEVL. The invention provides experimental basis for the application of the polypeptide in coated balloons and coated stents. Compared with traditional coating drugs, as a small fragment of Mfn2 itself, the polypeptide does not have the toxic side effects and immunogenicity of chemical drugs, and its practicability is better than rapamycin, so it has a wide range of application prospects and a large clinical application value.

Description

technical field [0001] The present invention relates to a polypeptide acting on vascular smooth muscle cells and its derivatives, in particular to a polypeptide inhibiting cell proliferation and inducing apoptosis, its preparation method and application. Background technique [0002] Atherosclerotic disease has become the number one killer of human health, and its incidence is increasing rapidly year by year, and the age of onset is getting younger. Severe coronary atherosclerotic lesions usually require percutaneous coronary intervention (PCI). At present, PCI has become a very important treatment for atherosclerotic diseases in China. In 2010, coronary stents were implanted in my country. The number has reached more than 300,000 cases, and intravascular stents have become the most commonly used intravascular implants. From the late 1980s to the early 1990s, Percutaneous transluminal coronary angioplasty (PTCA) was an important means of treating coronary heart disease. How...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C07K1/06C07K1/04A61L31/08A61L31/16
CPCY02P20/55
Inventor 郭小梅糜涛钱翠平白静刘纯刘晓雯彭稳中陈光慧
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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