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Application of circUTRN in preparation of medicine for treating heart failure, recombinant vector and medicine for treating heart failure

A recombinant carrier and heart failure technology, applied in the field of biomedicine, can solve the problems of bleeding at the puncture point, achieve good therapeutic effect, reduce the infarct size, and inhibit the effect of cardiomyocyte apoptosis

Pending Publication Date: 2022-05-06
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the treatment process often causes various complications, such as bleeding, air embolism, bleeding at the puncture site, infection, etc.

Method used

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  • Application of circUTRN in preparation of medicine for treating heart failure, recombinant vector and medicine for treating heart failure
  • Application of circUTRN in preparation of medicine for treating heart failure, recombinant vector and medicine for treating heart failure
  • Application of circUTRN in preparation of medicine for treating heart failure, recombinant vector and medicine for treating heart failure

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A recombinant vector expressing circUTRN is constructed by the following steps:

[0036] 1) Reverse transcribe the total RNA of the mouse heart tissue, perform PCR amplification sequentially, double enzyme digestion, and recover the cDNA sequence (as shown in SEQ ID NO.1) to obtain circUTRN; the nucleoside of the upstream primer of the PCR amplification The acid sequence is as shown in SEQ ID NO.2, specifically as follows: 5'-CGGAATTCTGAAATATGCTATCTTACAGTGGATCTCTTAGAGCTGAATACGACG-3', and the nucleotide sequence of the downstream primer for PCR amplification is as shown in SEQ ID NO.3, specifically as follows:

[0037] 5'-GGAATTCCATATGTCAAGAAAAAATATATTCACCGGTATGCAGTATTCTACCATCGG-3';

[0038] Each 50 μL PCR amplification reaction system consists of the following components: 1 μL of upstream primer (10 μM) (final concentration is 0.2 μM), 1 μL of downstream primer (10 μM) (final concentration is 0.2 μM), 2×TransStart FastPfuPCR SuperMix (full format Gold Biotechnology, Ca...

Embodiment 2

[0048] A recombinant vector expressing circUTRN similar to Example 1, the difference is:

[0049] 1. Replace the downstream primer in step 1) with a downstream primer whose nucleotide sequence is shown in SEQ ID NO.4, specifically as follows:

[0050] 5'-CGGGATCCAGTTGTTCTTACCGGTATGCAGTATTCTACCATCGG-3'; 2, replace NdeI (star selection enzyme) in step 1) with BamHI-HF (star selection enzyme);

[0051] 3. Replace the circular vector in step 2) with the AAV overexpression vector pK5ssAAV-ciR (purchased from Guangzhou Jisai Biotechnology Co., Ltd., catalog number GS0109).

Embodiment 3

[0053] A kind of medicine for treating heart failure, the preparation method of described medicine is as follows:

[0054] Use 293T cells to add the packaging system at an appropriate cell density (80%); add 1 mL of the packaging system to each 10 cm dish: 10 μg AAV2 / 9 (purchased from Addgene Plasmid, catalog number #112865), 10 μg pAdDeltaF6 plasmid (purchased from Addgene Plasmid, Cat. No. #112867), 10 μg of the recombinant vector constructed in Example 2, add DMEM medium (no FBS and no double antibody) to 910 μl, add 90 μl PEI transfection reagent (total volume: 1 mL); collect the supernatant and its Cells; use iodixanol to concentrate and purify the virus by ultracentrifugation (after collecting the supernatant, use a concentration column (Merck UFC905096) at 4000rpm 4°C to concentrate all the supernatant by centrifugation to 10-15mL), and the obtained virus suspension is as described Drug; the titer of the virus suspension is 1×10 13 vg / mL.

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Abstract

The invention relates to the technical field of biomedicine, in particular to application of circUTRN in preparation of a medicine for treating heart failure, a recombinant vector and the medicine for treating heart failure. Experiments prove that the medicine for treating heart failure prepared from circUTRN can treat or improve heart failure, and especially has a good treatment effect on heart failure caused by ischemia reperfusion injury; specifically, the circUTRN has the effect of improving cardiac systolic insufficiency caused by myocardial ischemia reperfusion injury for 3 weeks; in addition, the circUTRN can also reduce the infarction area of the acute myocardial ischemia reperfusion injury; overexpression of the circUTRN can inhibit myocardial cell apoptosis induced by oxygen-glucose deprivation / recovery.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to the application of circUTRN in the preparation of medicines for treating heart failure, recombinant vectors and medicines for treating heart failure. Background technique [0002] Heart disease is extremely harmful. Almost all cardiovascular diseases will eventually lead to heart failure. Myocardial damage caused by any cause such as myocardial infarction, cardiomyopathy, hemodynamic overload, and inflammation can cause myocardial structure and Changes in function eventually lead to poor ventricular pumping and / or filling function, which seriously threatens human life. Among them, ischemic cardiomyopathy is one of the major diseases that endanger the national health of our country. Timely reperfusion is a necessary step to save the ischemic heart, but this process is accompanied by severe myocardial ischemia / reperfusion (I / R) injury , there is still a lack of effective inter...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61P9/04C12N15/867C12N15/864
CPCA61K31/7088A61P9/04C12N15/86C12N2740/15043C12N2750/14143C12N2800/107Y02A50/30A61K31/713C12N2740/16043A61P9/00C12N15/111C12N2750/14122C12N2830/008
Inventor 肖俊杰王丽君胡玉雪孙江鹏陈宏健
Owner SHANGHAI UNIV
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