Application of MBOAT1 gene in preeclampsia

A preeclampsia, genetic technology, applied in the field of biomedicine, can solve the problems of mother and child harm, not very clear and so on

Active Publication Date: 2018-11-23
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the etiology and pathological mechanism of preeclampsia have not been fully elucidated, and no ideal method to completely cure the disease has been found clinically. The most thorough treatment method at present is to terminate pregnancy early, which causes great harm to mother and child.
Therefore, the exploration of the etiology and pathological mechanism of preeclampsia has always been an important topic of obstetrics research, and its pathogenesis is multifactorial, and it is not yet very clear

Method used

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  • Application of MBOAT1 gene in preeclampsia
  • Application of MBOAT1 gene in preeclampsia
  • Application of MBOAT1 gene in preeclampsia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening for Abnormally Expressed Genes in the Placental Tissue of Patients with Preeclampsia

[0054] 1. Sample collection

[0055] The placenta tissues of 43 cases of preeclampsia pregnant women and normal pregnant women were collected respectively, rinsed with normal saline twice, after removing water, they were divided into cryopreservation tubes, and stored at -80°C for later use.

[0056] Placental tissue excluded multiple pregnancy, infectious diseases, chemical drug dependence, maternal smoking, fetal congenital malformations and other pregnancy complications and complications, and all included subjects signed an informed consent form before collecting specimens. All the above specimens were obtained with the consent of the organizational ethics committee. Six samples were taken from each group for detection and analysis of gene expression profiles, screening of differentially expressed genes, and verification experiments were carried out in all 43 s...

Embodiment 2

[0071] Example 2 QPCR sequencing to verify the differential expression of the MBOAT1 gene

[0072] 1. Large-sample QPCR verification of differential expression of MBOAT1 gene.

[0073] 2. RNA extraction

[0074] Use QIAGEN tissue RNA extraction kit to extract total RNA in embryonic tissue, please refer to the manual for specific steps.

[0075] 3. Reverse transcription:

[0076] 1) Add dNTP mixture 1μl, Oligo dT primer 1μl, total RNA 2μg, add RNase FreeddH 2 O Make the total volume to 10 μl, carry out denaturation and annealing reaction on the PCR instrument, 65°C, 5min, place at 4°C after the reaction is completed.

[0077] 2) Construct a 20 μl reaction system, continue to add 4 μl of 5×Primer Script Buffer, 0.5 μl of RNase Inhibitor, 0.5 μl of Prime Script RTase, RNase Free dd H 2 O 5.0 μl, carry out the reverse transcription reaction on the PCR instrument according to the following conditions: 42°C for 15-30 minutes, 95°C for 5 minutes, after the reaction is completed, ...

Embodiment 3

[0095] Example 3 Western blot detection of the expression level of MBOAT1 protein

[0096] 1. Protein sample preparation

[0097] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0098] 2. Electrophoresis

[0099] β-actin was used as internal reference. 50 μg of total protein was separated by SDS-PAGE, electrotransferred to PVDF membrane, and blocked with 1× TBST containing 5% skimmed milk powder at room temperature for 1 hour; added MBOAT1 monoclonal antibody and β-actin monoclonal antibody respectively, and overnight at 4°C; 1× Wash the membrane 4 times with TBST, add secondary antibody, and incubate at room temperature for 1 h; wash the membrane 4 times with 1×TBST, react in Super Signal chemiluminescent reagent for 2 min, expose the X-ray film in a dark room, and develop and fix it by conventional methods.

[0100] 3. Statistical processing

[0101] The gray value of the protein band was analyzed using Image J so...

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Abstract

The invention discloses application of a molecular marker in preparing a drug for diagnosing and treating preeclampsia, and relates to the application of an MBOAT1 gene in preeclampsia. Content of theMBOAT1 gene and an expression product thereof in a placenta tissue of a subject can be detected to judge whether the subject has the risk of suffering from the preeclampsia or not or to diagnose whether the subject has the risk of suffering from the preeclampsia or not. Besides, proliferation indexes of in-vitro cultured placental trophoblast cells are researched to prove that the MBOAT1 gene canbe used as a drug target for treating preeclampsia.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of a molecular marker in the preparation of drugs for diagnosis and treatment of preeclampsia, more specifically to the application of MBOAT1 gene in preeclampsia. Background technique [0002] Hypertensive disorder complicating pregnancy (HDCP) is one of the common complications during pregnancy, which refers to transient hypertension (systolic blood pressure ≥ 140mm Hg (18.7k Pa) and / or Diastolic blood pressure ≥ 90mm Hg (12.0k Pa)), edema, proteinuria and other clinical symptoms, which disappear after delivery. Mother and child cause harm, and even lead to the death of mother and child. According to Williams Obstetrics, HDCP can be subdivided into subtypes such as pregnancy-induced hypertension, preeclampsia (mild and severe), eclampsia, chronic hypertension combined with preeclampsia, and pregnancy combined with chronic hypertension. Preeclampsia (PE) is the most com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883G01N33/6893G01N2800/368
Inventor 孙耀兰张冬梅
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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