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Application of cmdl-1, kit for diagnosing heart disease and drug for treating heart disease

A CMDL-1 and kit technology, applied in the field of biomedical engineering, can solve problems such as improving disease prognosis

Active Publication Date: 2022-07-05
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When congestive heart failure occurs, the mortality rate is approximately 50% and there are currently no treatments available to improve the prognosis of the disease

Method used

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  • Application of cmdl-1, kit for diagnosing heart disease and drug for treating heart disease
  • Application of cmdl-1, kit for diagnosing heart disease and drug for treating heart disease
  • Application of cmdl-1, kit for diagnosing heart disease and drug for treating heart disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 Changes in the expression level of CMDL-1 under the stimulation of doxorubicin (Dox)

[0086] 1.H 9 C 2 Cardiomyocyte culture

[0087] H 9 C 2 Cells were derived from rat cardiomyocytes in DMEM containing 10% serum, 1% penicillin and streptomycin in 5% CO. 2 in a cell incubator at 37°C.

[0088] 2. Dox treatment of cardiomyocytes

[0089] After adding Dox to culture, the total RNA of cells was extracted, and the expression level of CMDL-1 was detected by real-time fluorescence quantitative PCR.

[0090] Specifically, total RNA was extracted with Trizol reagent, and after DNase I (Takara, Japan) treatment, RNA was reverse transcribed into cDNA by reverse transcriptase. Quantitative real-time PCR was performed on a real-time PCR detection system (CFX96, Bio-Rad). The qRT-PCR system included 12.5 μl of 2×SYBR Green (Takara, Japan), 1 μl of primers (R / F), 1 μl of cDNA, and water to a total volume of 25 μl. lncRNA expression levels were quantified accordin...

Embodiment 2

[0091] Example 2 CMDL-1 can inhibit doxorubicin (Dox)-induced mitochondrial fission at the cellular level

[0092] 1.H 9 C 2 Cardiomyocyte culture

[0093] H 9 C 2 Cells were derived from rat cardiomyocytes in DMEM containing 10% serum, 1% penicillin and streptomycin in 5% CO. 2 in a cell incubator at 37°C.

[0094] 2. To H 9 C 2 Cardiomyocytes were transfected with CMDL-1 overexpression vector.

[0095] Specifically: prepare a density of 3 to 5 × 10 in complete medium 4 H / ml 9 C 2 Cell suspension, take appropriate cells to inoculate into culture plates, and culture in a cell culture incubator with 5% carbon dioxide for 16-24 hours until cell confluence is 20-30%. According to the cell infection multiplicity and virus titer, add the corresponding virus amount, the calculation formula is: virus volume = (infection multiplicity × cell number) / virus titer, after culturing in a 5% carbon dioxide cell incubator for 12-16 hours, replace it with Conventional medium, con...

Embodiment 3

[0099] Example 3 CMDL-1 can inhibit doxorubicin (Dox)-induced apoptosis at the cellular level

[0100] 1.H 9 C 2 Cardiomyocyte culture

[0101] H 9 C 2 Cells were derived from rat cardiomyocytes in DMEM containing 10% serum, 1% penicillin and streptomycin in 5% CO. 2 in a cell incubator at 37°C.

[0102] 2. To H 9 C 2 Cardiomyocytes were transfected with CMDL-1 overexpression vector.

[0103] Specifically: prepare a density of 3 to 5 × 10 in complete medium 4 H / ml 9 C 2 Cell suspension, take appropriate cells to inoculate into culture plates, and culture in a cell culture incubator with 5% carbon dioxide for 16-24 hours until cell confluence is 20-30%. According to the cell infection multiplicity and virus titer, add the corresponding virus amount, the calculation formula is: virus volume = (infection multiplicity × cell number) / virus titer, after culturing in a 5% carbon dioxide cell incubator for 12-16 hours, replace it with Conventional medium, continue to cul...

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Abstract

The invention provides an application of CMDL-1, a kit for diagnosing heart disease and a medicine for treating heart disease, and relates to the technical field of biomedical engineering. The long-chain non-coding RNA CMDL-1 related to heart disease provided by the invention, Its cDNA sequence is shown in SEQ ID NO.1. The inventors found through experiments that the expression of CMDL-1 was significantly down-regulated in cardiomyocytes stimulated by doxorubicin, and the purpose of diagnosing cardiac diseases can be achieved by specific detection of CMDL-1. At the same time, the overexpression of CMDL-1 can effectively inhibit the mitochondrial fission of cardiomyocytes induced by doxorubicin, and effectively reduce the apoptosis of cardiomyocytes induced by doxorubicin. It is suggested that CMDL-1 is expected to provide meaningful biological indicators for the diagnosis and treatment of clinical heart diseases, and provide new targets and new ideas for the design of anti-cardiac diseases.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to an application of CMDL-1, a kit for diagnosing heart disease and a medicine for treating heart disease. Background technique [0002] The anthracycline anticancer drug doxorubicin is an effective and commonly used chemotherapy drug for various malignant tumors. Use is limited due to cardiotoxic side effects. Doxorubicin-induced cardiomyopathy is a fatal disease that can be acute, generally occurs within 2-3 days of administration, and occurs in approximately 11% of patients. Acute left ventricular (LV) failure is a rare manifestation of acute cardiotoxicity that is reversible with appropriate treatment. The incidence of chronic cardiotoxicity due to doxorubicin is much lower, about 1.7%, and it usually occurs around 30 days after dosing, but can also occur within 6-10 years of treatment. The incidence of doxorubicin-induced cardiomyopathy is primarily dose-relat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68A61K38/17A61P9/00A61P9/10A61P9/04
CPCC12Q1/6883G01N33/6893A61P9/00A61P9/10A61P9/04C12Q2600/158G01N2333/47G01N2800/324G01N2800/325A61K38/00
Inventor 李培峰曾焕玉陈夏添
Owner QINGDAO UNIV
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