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TMEM16A taken as osteoporosis marker as well as application, osteoporosis diagnostic kit and medicine thereof

A diagnostic kit and osteoporosis technology, applied in the field of disease markers, can solve the problems of multi-observation data, inconvenient movement of ultrasound equipment, and inability to compare with the real value

Active Publication Date: 2019-12-20
HEBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing methods for diagnosing osteoporosis mainly have the following disadvantages: 1. The doctor's inquiry of past medical history can only provide reference, and cannot accurately determine whether he has osteoporosis
2. Bone mass measurement is a commonly used method nowadays, but whether a fracture occurs is mainly determined by bone strength, and bone strength is composed of many aspects such as bone mass, bone quality, bone composition, and bone shape. Quantitative judgment is not accurate enough
3. The result of ultrasonic measurement is not bone density or bone mineral content, which cannot be compared with the real value, and requires more observation data. In addition, the ultrasonic equipment is huge and inconvenient to move, and the cost of each measurement is high.

Method used

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  • TMEM16A taken as osteoporosis marker as well as application, osteoporosis diagnostic kit and medicine thereof
  • TMEM16A taken as osteoporosis marker as well as application, osteoporosis diagnostic kit and medicine thereof
  • TMEM16A taken as osteoporosis marker as well as application, osteoporosis diagnostic kit and medicine thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0051] In this example, the expression level of TMEM16A in osteoclasts of osteoporosis patients and normal people was measured. A minimal amount of bone tissue was extracted from osteoporosis patients through minimally invasive surgery, and primary osteoclasts were isolated from it, using complete medium (DMEM-α medium: fetal bovine serum=9:1) for cell culture. Through PCR experiments, the RNA content of TMEM16A in osteoclasts of patients with osteoporosis was analyzed compared with that of normal people. Using TMEM16A antibody to carry out cell immunoblotting experiment, compare and analyze the expression of TMEM16A in osteoclasts of patients with osteoporosis and the expression of normal people. The result is figure 1 As shown, it was found that the expression of TMEM16A in the bone tissue of patients with osteoporosis was about 1.5 times higher than that of normal people. Therefore, the increased expression of TMEM16A is a marker of osteoporosis.

Embodiment 2

[0053] In this implementation, a mouse osteoporosis model was first established, and then the TMEM16A content of the bone tissue of the osteoporotic mice was measured.

[0054] Mouse osteoporosis model establishment method: select female C57 mice aged 3-6 months, and the female mice will be artificially removed from the female mouse ovaries by surgery, and cultured for four weeks to build a model to simulate the female postmenopausal bone caused by lack of estrogen Poor texture; male mice are selected male C57 mice aged 2-4 months. The testes of female mice are artificially removed by surgery, and the model is cultured for four weeks to simulate the senile bone caused by the drastic reduction of androgen secretion in elderly men. Porosity.

[0055] The measurement methods of TMEM16A expression mainly include: measuring the changes in the RNA content of TMEM16A by PCR, including: Trizol extraction of total RNA, reverse transcription reaction, PCR reaction, and electrophoresis experi...

Embodiment 3

[0058] In this example, osteoclasts were induced and differentiated in vitro. The method of osteoclasts induced differentiation is as follows: bone marrow cells are extracted from 6-8 weeks old C57 mouse bone marrow, the red blood cell lysate is fully lysed and centrifuged to extract bone marrow stromal cells and add 10ng / Cultivate with ml of M-CSF, draw the supernatant after 24 hours, centrifuge at 1500 rpm, add medium containing 50ng / ml Rankl and 30ng / ml M-CSF and continue culturing for 5 days. Change the medium once during this period and use it after 5 days TRAP staining kit detects whether osteoclasts are successfully induced. The expression level of TMEM16A was detected by PCR and immunoblotting experimental methods, and the experimental methods were the same as the above examples.

[0059] Then detect the content of TMEM16A in cells 1, 3 and 5 days after normal induction, and the results are as follows image 3 with Figure 4 As shown, after 1, 3, and 5 days of normal ind...

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Abstract

The invention provides TMEM16A taken as an osteoporosis marker as well as an application, an osteoporosis diagnostic kit and a medicine thereof and relates to the technical field of disease markers. The osteoporosis diagnostic kit taking TMEM16A as the osteoporosis marker judges whether a subject suffers from osteoporosis by detecting content of TMEM16A in peripheral blood monocytes of the subject. Diagnosis is fast and accurate, operation is simple, and no special instrument or equipment is needed. The medicine used for treating the osteoporosis takes TMEM16A as a target and allievates osteoporosis symptons of a patient by regulating and controlling expression quantity of TMEM16A in the patient.

Description

Technical field [0001] The present invention relates to the technical field of disease markers, in particular to a TMEM16A as a marker of osteoporosis and its application, an osteoporosis diagnostic kit and a medicine. Background technique [0002] Osteoporosis is a systemic bone metabolism disease. Its main characteristics are decreased bone mass and changes in bone microstructure, accompanied by decreased bone strength, increased bone fragility, and increased fracture risk. According to epidemiological statistics, the prevalence of osteoporosis among people over 50 years old in my country is 20.7% for women and 14.4% for men. And according to global epidemiological surveys, the prevalence of osteoporosis is relatively high in all countries. The incidence of osteoporosis is related to many factors, among which gender factors and age factors are the most relevant to the occurrence of osteoporosis. Pain and fractures caused by osteoporosis seriously affect the quality of life of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68A61K31/7105A61P19/10
CPCA61K31/7105A61P19/10G01N33/6893
Inventor 安海龙李英贤陈娅斐凌树宽郭帅孙维佳王徐朝展永
Owner HEBEI UNIV OF TECH
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