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Application of SIRT (Silent Mating Type Information Regulation 2 Homolog 1) to prepare medicine used for regulating down the expression of cyclin D1

A 1. SIRT1, cell cycle technology, applied in drug combinations, peptide/protein components, anti-tumor drugs, etc., can solve problems such as the role of SIRT1 that has not been involved

Inactive Publication Date: 2011-04-13
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing studies have not addressed the role of SIRT1 in the regulation of cyclin D1 and related cell cycle proteins

Method used

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  • Application of SIRT (Silent Mating Type Information Regulation 2 Homolog 1) to prepare medicine used for regulating down the expression of cyclin D1
  • Application of SIRT (Silent Mating Type Information Regulation 2 Homolog 1) to prepare medicine used for regulating down the expression of cyclin D1
  • Application of SIRT (Silent Mating Type Information Regulation 2 Homolog 1) to prepare medicine used for regulating down the expression of cyclin D1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Construction of smooth muscle-specific human SIRT1 transgenic mice

[0046] 1. Cloning construction of SMC-SIRT1 for preparation of transgene

[0047] The mouse SM22α promoter containing a 5' wobble sequence of 445 base pairs is a vascular smooth muscle cell-specific promoter, which is only active in the aorta and middle arteries of adult rats, but in vein or visceral smooth muscle cells, myocardial or skeletal muscle cells were not expressed.

[0048] The mouse SM22α promoter fragment containing BssH II and BamHI restriction sites was ligated with the human wild-type SIRT1 plasmid double-digested with BamHI and NotI with T4 DNA ligase and cloned into a vector containing PolyA. For Escherichia coli cells, the single and combined enzyme digestion of the small plasmid was identified and sent for sequencing. The results obtained were confirmed by BLAST comparison on the Internet and the clone was named SMC-SIRT1. Related results see figure 1 .

[0049] 2. Pre...

Embodiment 2

[0075] Example 2 Mouse Common Carotid Artery Ligation Model Causes Neointimal Formation

[0076] Tribromoethanol (30mg / kg-50mg / kg) was injected intraperitoneally to anesthetize male C57BL / 6 mice at 12-14 weeks. The limbs and head of the mouse were fixed in the supine position, so that the neck was completely exposed. After the skin was disinfected, a midline incision was made on the upper part of the neck. Cut the skin and muscle layer longitudinally, with an opening of about 1-1.5cm. Gently separate the thyroid gland with straight and curved ophthalmic forceps, tear the sheath at the lower right, and the left common carotid artery with blood flow can be seen. Gently pick up the left common carotid artery with a small elbow hemostat, and separate the left common carotid artery from the accompanying vagus nerve with a small elbow hemostat on the other side, and further separate it to the bifurcation of the left common carotid artery. Near the bifurcation, the left common caro...

Embodiment 3

[0081] Expression of PCNA in embodiment 3 transgenic mice

[0082] As an indicator reflecting active cell proliferation, cell proliferating nuclear antigen (PCNA) is widely used. In order to further verify that SIRT1 can significantly inhibit neointima formation caused by common carotid artery ligation in vivo, SIRT1 immunohistochemical staining was first used to confirm the expression of SIRT1 in the arterial medial smooth muscle of transgenic mice ligated for 28 days, and then compared the ligated 28 PCNA immunohistochemical staining and PCNA staining index of day-old WT and SIRT1 transgenic mice. Compared with WT mice in smooth muscle-specific expressing SIRT1 transgenic mice, the number of PCNA-positive staining (tan or brown-yellow) cells in SIRT1 transgenic mice was significantly reduced, and the rate of PCNA-positive cells in the inner membrane was also significantly reduced (P Figure 8 .

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Abstract

The invention relates to the application of SIRT1 (Silence Information Regulator Tuin1) to prepare medicine used for regulating down the expression of cyclin D1. Particularly, the medicine is of a delegiert protein type, and the form of the medicine can be any form suitable for the application of protein medicine. The medicine is a reorganization nucleic acid construction body capable of expressing SIRT1 protein. The expression carrier of the medicine can be any expression carrier which is suitable for gene therapy and comprises a virus carrier, such as a reorganization retrovirus carrier, an adenovirus carrier, an adeno-associated virus carrier, a herpes simplex virus carrier, a vaccinia virus carrier, and the like and a nonviral carrier, such as a bacteriophage carrier, a replicon carrier, and the like.

Description

technical field [0001] The present invention relates to the use of SIRT1 in the preparation of medicines for regulating the expression of cell cycle proteins, in particular, the use of SIRT1 in the preparation of medicines for down-regulating the expression of cyclin D1, cyclin E or CDK2. Background technique [0002] Neointimal formation is closely related to vascular remodeling, mainly including the proliferation, migration, apoptosis of vascular smooth muscle cells, and the synthesis, degradation, and rearrangement of matrix components. Excessive vascular smooth muscle cell proliferation is an important pathological basis of cardiovascular diseases such as atherosclerosis, pulmonary hypertension, systemic hypertension, abdominal aortic aneurysm and restenosis after percutaneous transluminal angioplasty. [0003] Whether cell proliferation is normal or not is closely related to cell cycle. The cell cycle is divided into G1 phase (DNA synthesis preparation phase), S phase ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/50A61K48/00A61P9/00A61P9/10A61P35/00
Inventor 刘德培李莉陈厚早高鹏张慧娜张庆军
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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