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Application of miRNA in preparation of medicaments and diagnostic reagents for treating primary aldosteronism adenoid tumors

A diagnostic reagent, aldosterone technology, applied in the field of medicine and biology

Inactive Publication Date: 2014-01-22
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the expression of miRNA in the adrenal tissue of PA and its mechanism of action in the development of the disease have not been reported yet.

Method used

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  • Application of miRNA in preparation of medicaments and diagnostic reagents for treating primary aldosteronism adenoid tumors
  • Application of miRNA in preparation of medicaments and diagnostic reagents for treating primary aldosteronism adenoid tumors
  • Application of miRNA in preparation of medicaments and diagnostic reagents for treating primary aldosteronism adenoid tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 128 patients diagnosed with primary aldosteronism (PA) and treated with surgery

[0018] I. Screening object selection criteria:

[0019] (1) JNC (Joint National Committee on the Prevention, Evaluation, and Treetment of High BLood Pressure) hypertension grade 2 (BP>160-179 / 100-109mmHg), grade 3 (BP>180 / 110mmHg);

[0020] (2) Refractory hypertension (combined use of 3 antihypertensive drugs, which must include diuretics, and each antihypertensive drug has reached the conventional therapeutic dose, and the blood pressure is still greater than 140 / 90mmHg);

[0021] (3) Spontaneous or diuretic-induced hypokalemia;

[0022] (4) Adrenal accidental tumor;

[0023] (5) Family history of premature hypertension or premature (less than 40 years old) family history of cerebrovascular accident;

[0024] (6) There are first-degree relatives of hypertensive patients with primary aldehyde.

[0025] II. Diagnosis of Selected Subjects

[0026] Aldosterone / renin (ARR): All subjects sto...

Embodiment 2

[0042] Get the total RNA obtained in Example 1 and carry out microRNA chip analysis

[0043] (1) RNA quality control detection

[0044] The extracted primary aldosteronism (PA) adenoma, nodular hyperplasia tissue and normal adrenal cortex tissue were sent to LC Sciences for RNA quality testing. Qualified RNA was used for the next step of microarray gene expression detection.

[0045] (2) µParaflo TM microRNA microarray gene expression experiment

[0046] The samples sent for chip testing included RNA from 6 cases of adenoma tissue, 6 cases of RNA from nodular hyperplasia tissue and 3 cases of RNA from normal adrenal cortex tissue. Microarray gene expression detection experiments were completed by LC Sciences. The specific experimental procedure is as follows: 10 μg of total RNA sample is required for microarray experiments. Total RNA was separated by YM-100 (Millipore) microcentrifuge filter column to obtain small RNAs with a length less than 300nt. Poly(A) polymerase is...

Embodiment 3

[0053] According to the analysis of miRNA expression difference of the two kinds of microarray results in the embodiment, miR-375, miR-7 and miR-29a / b / c expression of adenoma, nodular hyperplasia tissue and normal adrenal cortex in primary aldosteronism The expanded sample verification of real-time PCR was carried out horizontally (the expanded samples included 6 cases of adenoma tissue, 6 cases of nodular hyperplasia tissue and 3 cases of normal adrenal cortex tissue that were originally sent for microarray detection). The result is as Figure 1-Figure 5 . The steps are:

[0054] I. Reverse transcription into cDNA

[0055]The operation of the reverse transcription process was carried out according to the instructions of the Exiqon Universal cDNA synthesis kit. This reverse transcription method can reverse transcribe all microRNA at one time for subsequent real-time PCR.

[0056] II. Real-time PCR

[0057] (1) Primer design and synthesis: both were completed by Exiqon. P...

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Abstract

The invention relates to the fields of medicaments and biotechnology, and in particular relates to a method for preparing and screening a medicament for treating primary aldosteronism adenoid tumors by using at least one of miR-375 or miR-7 according to a fact that different expression levels of miRNA in the primary aldosteronism adenoid tumors and hyperplastic tissues and the expression level of miR-375 are related to the tumor size of aldosteronoma, and a method for using a detection reagent of miR-375 or miR-7 to prepare a diagnostic reagent of the primary aldosteronism adenoid tumors.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to using miRNA to prepare and screen drugs and diagnostic reagents for treating primary aldosteronism adenoma. Background technique [0002] In April 1954, Jerome W. Conn, a professor of medicine at the University of Michigan, faced a female patient with high blood pressure, severe hypokalemia and obvious neuromuscular symptoms, who was diagnosed as "potassium-losing nephritis". In his pathophysiological research, he reported the world's first primary aldosteronism (primary aldosteronism), and then the aldosterone-secreting adrenal tumor disease was also named Conn's syndrome. In 1957, Professor Kuang Ankun and Professor Chen Jialun of Shanghai Ruijin Hospital diagnosed and reported the first case of primary aldehyde in China. In the past few decades, primary aldolism has been considered a rare adrenal disease, and the prevalence may be less than 1% to 2% in the hypertens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K48/00A61P35/00
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 宁光何娟王卫庆李小英曹亚南
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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