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Application of Bmal1 gene in preparation of product for detecting and treating cartilage hypoplasia disease

A kind of achondroplasia and genetic technology, applied in the field of biomedicine, can solve the problems of lack of detection and treatment, and unsatisfactory curative effect

Inactive Publication Date: 2021-09-10
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for achondroplasia, the current treatment methods cannot achieve satisfactory curative effect, and at the same time, there is a lack of drugs and detection methods developed for the Bmal1 gene and its target genes to detect and treat achondroplasia diseases

Method used

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  • Application of Bmal1 gene in preparation of product for detecting and treating cartilage hypoplasia disease
  • Application of Bmal1 gene in preparation of product for detecting and treating cartilage hypoplasia disease
  • Application of Bmal1 gene in preparation of product for detecting and treating cartilage hypoplasia disease

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Experimental program
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Effect test

Embodiment 1

[0023] We downloaded the expression profile chips (GSE145821) of achondroplasia mice and their control mice through the GEO database, extracted and further analyzed the expression of the biological clock gene Bmal1, and found that in the femur tissues of 1-4 week achondroplasia mice, Compared with the control group, the expression of Bmal1 decreased, suggesting that there is a correlation between the decrease of Bmal1 gene and achondroplasia (see figure 1 ).

Embodiment 2

[0025] We bred Bmal1-knockout mice, and compared them with littermate wild-type mice, we found that Bmal1-knockout mice were shorter than control mice at embryonic day 18.5 (see figure 2 ), the results of Masson staining showed that the amount of collagen fibers in the femoral metaphysis cartilage tissue of Bmal1 knockout mice decreased, and the results of transmission electron microscopy showed that the endoplasmic reticulum in the femoral metaphysis chondrocytes of Bmal1 knockout mice was significantly expanded swelling, suggesting that the formation of cartilage matrix is ​​regulated by Bmal1 (see image 3 , Figure 4 ).

Embodiment 3

[0027] We further extracted primary chondrocytes, knocked out Bmal1 in chondrocytes by infecting viruses, and detected the co-localization of chondrocyte type II collagen and endoplasmic reticulum by immunofluorescence experiments. The specific steps are as follows:

[0028] (1) Primary chondrocyte culture

[0029] 1) SD male rats, 60-80g, 2-3 weeks old, were killed by neck dislocation, and soaked in alcohol for 15 minutes;

[0030] 2) Take out the rat femur, carefully separate the femoral metaphyseal cartilage, and soak it with PBS;

[0031] 3) Transfer the cartilage tissue to an EP tube and cut it into pieces, add 1.5 mL of 0.25% trypsin, mix well, and place it in a 37°C water bath for 30-45 minutes to digest;

[0032] 4) Centrifuge the digested cartilage tissue at 1000 rpm for 5 minutes, discard the supernatant, add 8 mL of 0.25% type II collagenase prepared with basal medium, resuspend in a 15 mL centrifuge tube, and digest in an incubator at 37°C for 8-12 hours;

[0033...

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Abstract

The invention discloses application of a Bmal1 gene in preparation of a product for detecting and treating cartilage hypoplasia diseases, and belongs to the technical field of biological medicines. The nucleotide sequence of the Bmal1 gene in the invention is as shown in SEQ ID NO. 1, and the amino acid sequence of the Bmal1 gene is as shown in SEQ ID NO. 2. According to the invention, a molecular mechanism of the Bmal1 gene for regulating and controlling the biological process of cartilage growth and development is clarified, and a foundation is laid for research and development of drugs taking the P4ha1 gene as a target based on the biological clock gene Bmal1.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and particularly relates to the application of Bmal1 gene in the preparation, detection and treatment of achondroplasia disease products. Background technique [0002] Achondroplasia (Achondroplasia) is the most common skeletal dysplasia disease that causes short stature, and its incidence rate is about 1:15000-25000. The etiology of achondroplasia has not been fully elucidated, but it is mainly caused by genetic factors and environmental factors. Achondroplasia seriously endangers the normal growth and development of individuals. Understanding the molecular pathology of achondroplasia is therefore extremely important for the prevention and treatment of the disease. The cartilage growth plate is located between the epiphysis and the diaphysis, as a scaffold for the deposition and mineralization of bone tissue, it determines the longitudinal growth of the bone. Cartilage tissue is mainly com...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/375A61P19/08C12Q1/6883
CPCA61K45/00A61K31/375A61P19/08C12Q1/6883C12Q2600/158
Inventor 陈莉莉陈广进唐清明尹盈孙纪威彭金枫
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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