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Application of LncRNAGAS5 to treatment on primary osteoporosis

A technology for osteoporosis and primary disease, applied in the field of molecular biomedicine, can solve the problems of decreased skin absorption, poor patient comfort, scar hyperplasia, etc., and achieve strong anti-osteoporosis ability, improved acceptance and comfort Effect

Inactive Publication Date: 2020-08-25
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its dose can be applied in a small range, and it is impossible to increase the dose to meet the needs of patients with different severities
Moreover, teriparatide is currently only available as an injection, and patients need to inject it every day, and it will be effective if it is used for a long time
This may lead to adverse reactions after long-term skin injection, such as scar hyperplasia, decreased skin absorption at the injection site, poor patient comfort, etc.

Method used

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  • Application of LncRNAGAS5 to treatment on primary osteoporosis
  • Application of LncRNAGAS5 to treatment on primary osteoporosis
  • Application of LncRNAGAS5 to treatment on primary osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 constructs the adenovirus of GAS5 overexpression plasmid

[0022] The target shuttle plasmid pAdeno-mCMV-GAS5-EGFP-3FLAG was co-transfected with the adenovirus backbone plasmid into HEK293 cells to recombine to obtain the virus Adeno-GAS5-EGFP-3FLAG, which was purified after massive amplification, and the titer and target gene expression were detected Standby after leveling. The specific steps of the experiment are as follows:

[0023] 1. Virus packaging and identification

[0024] 1. The day before transfection, inoculate the cells into a 6-well plate, and control the density of the cells to be 70%-80% during transfection.

[0025] 2. Take out the cell culture plate one hour before transfection, take out the original cell culture medium, add 1.5ml of Opit-MEM medium, the total volume is 250ul, and mix gently.

[0026] 3. Prepare the complex of the transfection kit plasmid:

[0027] A. Dissolve 4ug of viral vector plasmid to be transfected (backbone pla...

Embodiment 2

[0062] Embodiment 2 animal experiments

[0063] 1. Construction of primary osteoporosis mouse model

[0064] Thirty C57 female mice were divided into 3 groups, normal growth + control virus group; osteoporosis mouse model + control virus group; osteoporosis mouse model + GAS5 overexpression adenovirus group. At the age of 12 weeks, the latter two groups were subjected to ovariectomy, and their ovaries were removed to establish a mouse model of primary osteoporosis.

[0065] 2. GAS5 overexpression adenovirus and control were used for in vivo injection.

[0066] Two months later, the adenovirus containing the GAS5 overexpression plasmid was injected through the tail vein using a microsyringe. 5 x 10 per mouse 11 The virus particles were dissolved in 200ul saline and injected into the tail vein.

[0067] 3. Detection of bone formation indexes and bone quality of mice in each group

[0068] After February and May, blood was collected from eyeballs, serum was collected, and se...

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Abstract

The invention provides application of long-chain non-coding RNAGAS5 to preparation of a medicament for treating skeleton system diseases. A GAS5 adenovirus is applied to carry out treatment on osteoporosis, and the pharmacological action of the GAS5 adenovirus is similar with that of teriparatide, and both the GAS5 adenovirus and the teriparatide are medicaments for promoting bone formation. The difference is that the GAS5 is naturally expressed in various types of cells in vivo, has no specific glandular secretion and cannot cause corresponding inhibition from endocrine glands of the human body; and the GAS5 only promotes MSCs to be differentiated into bone cells in vivo, and has no obvious influence on osteoclast. Therefore, the long-chain non-coding RNAGAS5 is higher in osteoporosis resistance, and has no similar problem of small dosage range of the teriparatide. In addition, due to the unique action mode of the adenovirus, the GAS5 can take an acting effect of resisting to osteoporosis only by injecting in vitro for once, and acceptability and comfort of a patient can be greatly promoted.

Description

technical field [0001] The invention relates to the field of molecular biomedicine, in particular to the role of LncRNAGAS5 in treating primary osteoporosis. Background technique [0002] Osteoporosis is the most common skeletal disease. It is a systemic bone disease characterized by low bone mass and damage to the microarchitecture of bone tissue, resulting in increased bone fragility and susceptibility to fracture. In 2001, the National Institutes of Health of the United States defined it as a skeletal disease characterized by decreased bone strength and increased fracture risk, suggesting that decreased bone mass is the main risk factor for osteoporotic fractures, but there are other risk factors. Osteoporosis can occur at any age, but is more common in postmenopausal women and older men. Osteoporosis is divided into primary and secondary two categories. Primary osteoporosis includes postmenopausal osteoporosis (type I), senile osteoporosis (type II) and idiopathic oste...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61K35/761A61P19/10C12Q1/6883
CPCA61K31/7105A61K35/761A61P19/10C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 沈慧勇李明谢中瑜吴燕峰王鹏李进腾郑冠刘文杰
Owner SUN YAT SEN UNIV
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