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Application of 11beta-HSD2 and epigenetics as early markers of articular cartilage dysplasia and osteoarthritis susceptibility

A technology for dysplasia and osteoarthritis, applied in the field of clinical medicine, which can solve problems such as inconvenience

Inactive Publication Date: 2018-03-23
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is worth noting that at present, the laboratories have not yet reached a consensus on the isolation and in vitro expansion of WJ-MSCs, and there is no method that can quickly and efficiently isolate homogeneous and well-defined WJ-MSCs. The implementation of clinical early warning based on high-throughput screening provides a lot of inconvenience

Method used

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  • Application of 11beta-HSD2 and epigenetics as early markers of articular cartilage dysplasia and osteoarthritis susceptibility
  • Application of 11beta-HSD2 and epigenetics as early markers of articular cartilage dysplasia and osteoarthritis susceptibility
  • Application of 11beta-HSD2 and epigenetics as early markers of articular cartilage dysplasia and osteoarthritis susceptibility

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Fetal cartilage of control group and PCE group, 12-week-old cartilage 11β-HSD2 promoter region H3K9ac, 11β-HSD2 and cartilage marker genes type II collagen (collagen type II alpha 1, COL2A1) and proteoglycan (aggrecan, AGAN) mRNA The expression level.

[0047] 1. Experimental animals

[0048] SPF grade healthy Wistar rats were purchased from Hubei Provincial Center for Disease Control and Prevention, animal license number: SCXK (E) 2008-2010. This study was approved by the Ethics Committee of Wuhan University Medical Center, and was carried out in strict accordance with the relevant treatment guidelines of international laboratory animal protection certification and evaluation institutions.

[0049] The experimental animals were kept in a barrier environment with a temperature of 22-25° C. and a humidity of 50%, with 12 hours of day and night alternation.

[0050] 2. Animal handling

[0051] Healthy and adult Wistar rats, weighing 209 ± 12 g for females and 258 ± 17 ...

Embodiment 2

[0066] The mRNA expression levels of 11β-HSD2 promoter region H3K9ac, 11β-HSD2 and downstream genes in normal control group and IUGR group.

[0067] 1. Umbilical cord mesenchymal stem cells WJ-MSCs cultured for total RNA extraction

[0068] Neonatal umbilical cord specimens were collected from the Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University. Divided into two groups: normal control group and IUGR group (n≧8), total RNA was extracted after culturing mesenchymal stem cells. Add 1ml Trizol to the homogenizer, after fully homogenized on ice, transfer to RNase-free 1.5ml EP tube; add 200μl chloroform for extraction, shake vigorously for 15s, let stand on ice for 10min; centrifuge at 12,000×rpm at 4°C for 15min; Carefully pipette 500 μl of the upper aqueous phase into a new RNase-free 1.5ml EP tube, add an equal volume of isopropanol, invert and mix to precipitate RNA, let stand at 20-30°C for 10 minutes; centrifuge at 12,000×rpm at 4°C for 10 minu...

Embodiment 3

[0080] Changes in the expression levels of H3K9ac, 11β-HSD2 in the umbilical cord 11β-HSD2 promoter region of neonates in the normal control group and IUGR group, as well as cartilage marker genes type II collagen (collagen type II alpha 1, COL2A1) and proteoglycan (aggrecan, AGAN).

[0081] 1. Detection of 11β-HSD2 expression and cartilage marker gene expression in neonatal umbilical cord tissue

[0082] Neonatal umbilical cord specimens were collected from the Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University. They were divided into two groups: normal control group and IUGR group (n≧8). A 10 cm umbilical cord was cut into pieces and added to culture medium containing (1 mg / mL) hyaluronidase collagenase I (Invitrogen). Shake at a constant speed at 37°C for 12 hours, add PBS solution and pipette repeatedly to dilute, collect the diluted solution (including tissue), centrifuge at 3500rpm for 20min, then discard the supernatant and floating adipose ...

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Abstract

The invention discloses the application of 11beta-HSD2 and epigenetics as early markers of articular cartilage dysplasia and osteoarthritis susceptibility. Research results show that in comparison with a normal control group, the level of histone acetylation and expression of the 11beta-HSD2 promoter region in the IUGR neonatal umbilical cord mesenchymal stem cells or the whole umbilical cord is decreased; and cartilage differentiation ability of the IUGR neonatal umbilical cord mesenchymal stem cells is lower, and cartilage degeneration similar to osteoarthritis is more likely to occur underthe action of glucocorticoids. Therefore, by detecting the expression and epigenetic levels of umbilical cord 11beta-HSD2 gene, the risk of cartilage dysplasia and osteoarthritis in adulthood is judged. The invention provides the research basis for the early warning technology of articular cartilage dysplasia and fetal osteoarthritis in adulthood, and also provides new ideas for comprehensive prevention and treatment of fetal osteoarthritis in adulthood.

Description

technical field [0001] The invention belongs to the technical field of clinical medicine, and relates to a type 2 11β-hydroxysteroid dehydrogenase (11β-hydroxysteroid dehydrogenase type-2, 11β-HSD2) gene and its epigenetics as articular cartilage dysplasia and early adult osteoarthritis Early warning biomarkers. Background technique [0002] Osteoarthritis is a chronic joint disease characterized by degeneration of articular cartilage. The traditional view is that osteoarthritis is a degenerative disease of the elderly. Because of its high incidence and difficult treatment, it is of great significance to study the diagnosis and treatment of osteoarthritis. Intrauterine growth retardation (IUGR) refers to a full-term baby whose birth weight is less than 2.5 kg, or a fetus whose weight is less than two standard deviations below the average weight for its gestational age. Large-scale epidemiological studies have found that low birth weight infants are susceptible to osteoart...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158
Inventor 汪晖陈廖斌齐勇建李斌黎伟
Owner WUHAN UNIV
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