lncRNA OGRU and drug use of lncRNA OGRU overexpression vector

A technology of long-chain non-coding and over-expression vectors, which is applied in the drug application field of disused osteoporosis drugs, long-chain non-coding RNAOGRU and its over-expression vectors, which can solve problems such as human health hazards and promote differentiation and mineralization ability, alleviating new bone formation barriers, good prospects for translational medicine

Active Publication Date: 2019-11-12
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing anti-osteoporosis drugs can effectively reduce the incidence of fractures in patients with osteoporosis, but these drugs will produce certain side effects, such as osteonecrosis, hypercalcemia and thromboembolic diseases, etc., which have a great impact on human health. big hazard

Method used

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  • lncRNA OGRU and drug use of lncRNA OGRU overexpression vector
  • lncRNA OGRU and drug use of lncRNA OGRU overexpression vector
  • lncRNA OGRU and drug use of lncRNA OGRU overexpression vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] To analyze the expression and characteristics of long non-coding RNA OGRU in 2D rotative MC3T3-E1 cell unloading model and hindlimb tail suspension (HLU) unloading osteoporosis mouse model, including the following steps:

[0049] Step 1. Establish a 2D rotary MC3T3-E1 cell unloading model

[0050] The MC3T3-E1 cells were subcultured into a rotary flask in an ultra-clean bench, and placed in a 37°C incubator for routine culture. After 6-8 hours of passage, the cells were completely adhered to the wall, and the spinner bottle was filled with α-MEM medium (Gibco, USA) containing 10% fetal bovine serum (HyClone, USA) and 1% double antibody (Invitrogen, USA), and stood upright. Place in a 37°C incubator overnight. On the second day, the air bubbles in the rotary bottle were completely exhausted in the ultra-clean bench. The gyratory group was installed on a 2D gyrator and rotated vertically with a gyration radius of 1.5 cm and a rotational speed of 24 rpm; the blank contro...

Embodiment 2

[0068] The OGRU overexpression vectors pcDNA3.1(+)-OGRU and siRNA-OGRU were transfected into MC3T3-E1 cells, and then their osteogenic differentiation and mineralization capabilities were detected, including osteogenic differentiation indicators (Alp, Osx, Runx2, Ocn) mRNA and protein expression, Alp activity, Alp staining and alizarin red staining etc., specifically comprise the following steps:

[0069] Step 1, Cell Transfection

[0070] Using Lipofectamine 3000kit (Invitrogen, USA), siRNA-OGRU or siRNA-NC (GenePharma, China; final concentration: 80nM), OGRU overexpression vector pcDNA3.1(+)-OGRU (GeneCreate, China; final concentration: 200ng / μL ) were respectively transferred into MC3T3-E1 cells, and a blank control group (CON) and an empty vector group (Vector) were set up, and subsequent experiments were performed as required. Among them, the sequences of siRNA-OGRU and si-NC are respectively shown in Table 3.

[0071] Table 3 siRNA sequence

[0072]

[0073] Step 2...

Embodiment 3

[0091] Investigate the effect of OGRU overexpression vector on the differentiation of MC3T3-E1 cells in the 2D rotary cell unloading model, including the following steps:

[0092] After MC3T3-E1 cells were transfected into the OGRU overexpression vector for 12 hours, they were placed in a 2D rotator to rotate for 48 hours, and then RNA and protein were extracted for subsequent qRT-PCR, Alp activity and western blot detection. The detection method was the same as in Example 2, and the detection results Respectively as Figure 9-11 shown.

[0093] Depend on Figure 9-11 It can be seen that the overexpression vector of OGRU can partially alleviate the differentiation disorder of MC3T3-E1 cells caused by 2D inversion.

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Abstract

The invention discloses lncRNA OGRU and drug use of an lncRNA OGRU overexpression vector. The expression of the lncRNA OGRU in a 2D rotation MC3T3-E1 cell de-load model and a mouse hind limb unweighted (HLU) de-load model is significantly reduced, the OGRU overexpression vector pcDNA3.1(+)-OGRU promotes cell differentiation of mouse preosteoblast MC3T3-E1 in the 2D rotation MC3T3-E1 cell de-load model, and pcDNA3.1(+)-OGRU is delivered to a mouse osteogenesis area by a bone targeted delivery system (DSS) 6-liposome to effectively relieve osteoporosis caused by HLU; and OGRU has potential to become a new target for disuse osteoporosis diagnosis biomarkers and treatment, and the OGRU overexpression vector is used in drugs for treating the disuse osteoporosis.

Description

technical field [0001] The invention relates to the field of molecular biomedicine, in particular to the pharmaceutical application of a long-chain non-coding RNA OGRU and its overexpression carrier, which is mainly used in disuse osteoporosis medicines. Background technique [0002] Osteoporosis is a systemic skeletal disease characterized by decreased bone mass and deterioration of bone microarchitecture leading to reduced bone strength and increased fracture risk. Globally, the incidence of osteoporotic fractures is highest in North America and Europe, followed by Asia, the Middle East, Oceania, Latin America, and Africa. Considering that Asia hosts most of the world's population, it is estimated that by 2050, more than 50% of the world's osteoporotic fractures will occur in Asia. [0003] Disuse osteoporosis is a common type of osteoporosis, the main cause of which is the long-term unloaded state of bones caused by long-term bed rest, motor paralysis, fixation after fra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6883A61K31/7105A61P19/10
CPCC12N15/113C12Q1/6883A61K31/7105A61P19/10C12N2310/10C12Q2600/158C12Q2600/178
Inventor 王可张舒石菲王艺璇胡泽兵曹新生张丽君李高志
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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