Application of Gs alpha gene in preparation of medicament for resisting abdominal aortic aneurysm

A technology for abdominal aortic aneurysm and aortic aneurysm, which is applied in the measurement/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of decreased expression of contractile proteins, stiffness and elevation of abdominal aortic wall, and achieves reduction The effect of small morbidity

Active Publication Date: 2019-04-19
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When an abdominal aortic aneurysm occurs, the vascular smooth muscle cells transform from contractile to synthetic, showing a decrease in the expression of contractile proteins, while the expression of synthetic proteins such as Vimentin and Osteopontin increases, and secretes MMP2 / MMP9, Collagen, and Fibronectin at the same time , resulting in stiffness of the abdominal aortic wall and decreased contractility

Method used

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  • Application of Gs alpha gene in preparation of medicament for resisting abdominal aortic aneurysm
  • Application of Gs alpha gene in preparation of medicament for resisting abdominal aortic aneurysm
  • Application of Gs alpha gene in preparation of medicament for resisting abdominal aortic aneurysm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The construction of embodiment 1 mouse abdominal aortic aneurysm model

[0030] Gsα flox / flox Mouse and smooth muscle-specific expression of Cre recombinase mouse SM22-CreER T2 Mating, get Gsα flox / flox / Cre + Mice, for five consecutive days to Gsα flox / flox / Cre + Mice were intraperitoneally injected with 1 mg of tamoxifen drug (Sigma Company) to stimulate the expression of Cre recombinase, thereby converting Gsα flox / flox / Cre + The sequence between the loxP sites in the mouse is recombined, that is, the Gsα flox / flox / Cre + The Gsα gene in the smooth muscle of the mouse is knocked out, and the function of the Gsα gene is inhibited, so as to obtain a mouse with a smooth muscle-specific knockout of the Gsα gene. The mouse is treated with Gsα SMKO express. figure 1 A strategy for conditional knockout of the Gsα gene.

[0031] On the 16th day after intraperitoneal injection of tamoxifen, Gsα SMKO The aorta of the mice was taken out, and at the same time, the c...

Embodiment 2

[0035] Example 2 Effect of smooth muscle Gsα gene on mouse abdominal aortic aneurysm

[0036] The double knockout mouse ApoE that embodiment 1 obtains - / - / Gsα SMKO and control mice without the ApoE of the Cre recombinase gene - / - / Gsα flox / flox Divided into four groups, with at least 6 mice in each group, processed at the age of 10 weeks. The mice in the four groups and their processing conditions were as follows:

[0037] Group 1: Control mouse ApoE - / - / Gsα flox / flox , pump in saline;

[0038] Group 2: Control mouse ApoE - / - / Gsα flox / flox , pumped into AngII;

[0039] The third group: double knockout mouse ApoE - / - / Gsα SMKO , pump in saline;

[0040] Group 4: double knockout mouse ApoE - / - / Gsα SMKO , pumped into AngII;

[0041] The operation procedure of pumping is as follows: Alzet micropump is subcutaneously implanted on the back of the mouse, and AngII is pumped in at a rate of 1000ng / Kg / min, or the same amount of normal saline is pumped in respectively,...

Embodiment 3

[0043] Example 3 Preparation of lentivirus against Gsα gene

[0044] 1. Construction of pFUGW-Gsα vector:

[0045] 1) The pFUGW plasmid (purchased from Addgene, #14883) was double-digested with endonucleases HindIII and KpnI, and the double-digested product was subjected to agarose gel electrophoresis, and the gel was cut and recovered;

[0046] 2) using the smooth muscle cell cDNA as a template, amplifying and purifying the Gsα gene fragment by PCR;

[0047] The specific primer nucleotide sequence for PCR amplification is as follows:

[0048] Forward primer: 5'-AAGCTTATGGGCTGCCTCGGGAACAGTA-3';

[0049] Reverse primer: 5'-GGTACCTTAGAGCAGCTCGTACTGACGAA-3';

[0050] The nucleotide sequence of the Gsα gene is shown in SEQ ID No.1;

[0051] 3) Digesting the Gsα gene fragment obtained in step 2) by endonucleases HindIII and KpnI to obtain a double digestion product of the Gsα gene;

[0052] 4) Perform agarose gel electrophoresis on the Gsα gene double-digestion product in step...

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Abstract

The invention relates to an application of a Gs alpha gene in preparing a medicament for resisting abdominal aortic aneurysm. According to the invention, an important role of a Gs alpha gene is playedon regulating the phenotype transformation of smooth muscle and maintaining the contraction function of the smooth muscle by constructing a gene knock-out mouse specifically knocking out the Gs alphaof the smooth muscle; meanwhile, by pumping AngII subcutaneously to construct a mouse abdominal aortic aneurysm model, that the Gs alpha gene knock-out mouse can significantly increase the incidenceof abdominal aortic aneurysm is discovered; in addition, according to the invention, a lentivirus of the Gs alpha gene is prepared, and the lentivirus of the Gs alpha gene can be injected into the tail vein of the mouse, so that the incidence rate of the abdominal aortic aneurysm of the mouse can be obviously reduced. According to the invention, the Gs alpha gene is used as an action target to prepare or screen a medicament for resisting the abdominal aortic aneurysm, and the medicament has important significance in the treatment of the abdominal aortic aneurysm.

Description

technical field [0001] The invention relates to the application of Gsα gene in the preparation of anti-abdominal aortic aneurysm medicine, which belongs to the technical field of medicine. Background technique [0002] Abdominal aortic aneurysm refers to a pathological state in which the local abdominal aorta shows tumor-like expansion, and the lumen diameter exceeds 50% of the normal value. It is a potentially fatal disease. Once the aneurysm ruptures, the mortality rate exceeds 80%. In the United States, the annual incidence of abdominal aortic aneurysm is about 20-40 people / 100,000 people, and about 15,000 people die of abdominal aortic aneurysm every year, accounting for the 13th cause of death. In my country, the incidence of abdominal aortic aneurysm The incidence rate of abdominal aortic aneurysm is also increasing year by year, and the incidence rate of abdominal aortic aneurysm detected by ultrasound in men over 65 years old is as high as 5%. Aneurysmal dilatation o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/106C12Q2600/178
Inventor 张文程秦小腾闫鹏
Owner SHANDONG UNIV QILU HOSPITAL
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