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Gene related to preeclampsia and application thereof

A technology of preeclampsia and gene markers, applied in the field of biomedicine, can solve problems such as abnormal remodeling of trophoblast spiral arteries, and achieve excellent specificity and sensitivity

Active Publication Date: 2022-05-13
THE AFFILIATED HOSPITAL OF QINGDAO UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of preeclampsia is still unclear, but most of the evidence has shown that insufficient trophoblast invasion leads to abnormal remodeling of spiral arteries, which in turn leads to the occurrence of preeclampsia.

Method used

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  • Gene related to preeclampsia and application thereof
  • Gene related to preeclampsia and application thereof
  • Gene related to preeclampsia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0049] Effects of inhibiting AC011753.5 on the proliferation ability of human trophoblast cell HTR-8 / Svneo

[0050] 1. Design and detection of siRNA for AC011753.5

[0051] (1) The sequence of si-AC011753.5 is shown below

[0052] 5'-UAGCAAAAUAUGCAAAUACAA-3', SEQ ID NO.6

[0053] 5'-GUAUUUGCAUAUUUUGCUAUA-3', SEQ ID NO.7;

[0054] (2) Set up 3 groups, namely the control group (idling lip2000), si-AC011753.5 group and si-NC group, and detect the relative expression of AC011753.5 after 48 hours of transfection;

[0055] Inhibition effects such as image 3 As shown, it can be seen that the expression level of AC011753.5 in the si-AC011753.5 group is 0.108±0.048, P<0.05, indicating that si-AC011753.5 can significantly inhibit the expression of AC011753.5.

[0056] 2. Detection of the effect of inhibiting AC011753.5 on the proliferation ability of human trophoblast cells HTR-8 / Svneo

[0057] (1) Human trophoblast cells HTR-8 / Svneo transfected with si-NC and si-AC011753.5 were see...

Embodiment 3

[0063] Effect of inhibiting AC011753.5 on the invasion ability of human trophoblast cell HTR-8 / Svneo

[0064] (1) Put the Matrigel gel at 4°C to melt into a liquid state, add serum-free medium to the Matrigel gel at a ratio of 5:1, mix well, add 100ul Matrigel gel to the upper chamber of the Transwell chamber, and put Incubate for 4 hours in the incubator;

[0065] (2) Human trophoblast cells HTR-8 / Svneo transfected with si-NC and si-AC011753.5 were digested with trypsin, washed with PBS, suspended in serum-free medium, and counted;

[0066] (3) Add 600ul medium containing 10% serum to the lower chamber of the Transwell chamber, add 100ul containing 2×10 serum to the upper chamber 4 The single-cell suspension of 10 cells was placed in a cell culture incubator for 24 hours;

[0067] (4) After the culture is over, take out the Transwell chamber with tweezers, blot the liquid in the upper chamber, place the Transwell in a cell culture well containing methanol, and fix at room t...

Embodiment 4

[0072] Detection of the effect of inhibiting AC011753.5 on the invasion-related proteins MMP5 and MMP9 of human trophoblast cells HTR-8 / Svneo

[0073] (1) Human trophoblast cells HTR-8 / Svneo were inoculated in 6-well plates, and transfected with si-NC and si-AC011753.5 respectively. After 48 hours of transfection, the medium was discarded, washed 3 times with PBS, and 200ul was added protein lysate;

[0074] (2) Use a cell scraper to scrape off the cells, transfer to a 1.5ml EP tube, and lyse on ice for 30 minutes;

[0075] (3) 4°C, 10000rpm / min, centrifuge for 15min, use a pipette to carefully absorb the supernatant to obtain the total cell protein;

[0076] (4) Detect the protein concentration according to the instructions of the BCA kit, add 5× loading buffer, place in boiling water for 5 minutes, and obtain the protein sample;

[0077] (5) Configure 10% stacking gel and 5% separating gel, and load the sample according to the electrophoresis tank;

[0078] (6) 80V consta...

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Abstract

The invention provides a gene related to preeclampsia and application thereof, and belongs to the technical field of biological medicine. The gene is AC011753.5, and the AC011753.5 is highly expressed in a placenta of a patient with preeclampsia. By inhibiting the expression of AC011753.5 in human trophoblasts, the proliferation and invasion ability of the trophoblasts can be obviously promoted. Therefore, the compound can be used for preparing the medicine for treating preeclampsia.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a gene related to preeclampsia and its application. Background technique [0002] Preeclampsia is a type of hypertensive disorder of pregnancy that is unique to pregnant women. Preeclampsia is a major cause of maternal and perinatal deaths and serious complications in my country. How to effectively diagnose and treat preeclampsia is still an urgent problem in obstetrics and gynecology. At present, the pathogenesis of preeclampsia is still unclear, but most of the evidence has shown that insufficient trophoblast invasion leads to abnormal remodeling of spiral arteries, which in turn leads to the occurrence of preeclampsia. [0003] Long non-coding RNA is a class of RNA with a length of more than 200nt nucleotides. Although long non-coding RNAs do not have the ability to encode, they can regulate gene expression from multiple levels such as epigenetic regulation,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11A61K31/713A61P15/00
CPCC12Q1/6883A61K31/713A61P15/00C12Q2600/158
Inventor 郑烨崔雪娜王琳
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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