Application of miR-135a in preparation of drugs for treating myocardial fibrosis

A technology of 1.mir-135a and 2.mir-135a, applied in the field of biomedicine, can solve the problem of no research on the influence of fibrosis-related pathways

Inactive Publication Date: 2018-10-16
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many studies on the regulatory factors of TRPM7, but there is no report on the regulatory effect of microRNA135a (miR-135a) on cardiac fibrosis by means of fibrosis regulation
[0006] At present, the research on miR-135a is mainly focused on tumor growth and nervous system lesions. The research results show that miR-135a participates in the regulation of apoptosis pathway by regulating different target genes, thereby affecting the survival of tumor cells, but miR-135a has no effect on cardiac fiber. The regulation of fibrosis has not been reported, and the effect on fibrosis-related pathways has not been studied

Method used

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  • Application of miR-135a in preparation of drugs for treating myocardial fibrosis
  • Application of miR-135a in preparation of drugs for treating myocardial fibrosis
  • Application of miR-135a in preparation of drugs for treating myocardial fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Detection of microRNA in plasma of patients with atrial fibrillation

[0045] 10 cases of plasma from healthy and simple atrial fibrillation patients were obtained from Jiangsu Provincial People's Hospital. The sample age was 50±5 years old. The experiment was approved by the Ethics Committee of Jiangsu Provincial People's Hospital and China Pharmaceutical University.

[0046] Medicines and reagents: DEPC water (Biyuntian Bio), miRNeasy Serum / Plasma kit (QIAGEN, Germany), miRNA first strand cDNA synthesis (tailing method) kit (Biotech), Premix Ex Taq TM II (Dalian Biotech)

[0047] 1. Extraction of plasma microRNA

[0048] (1) Plasma sample processing

[0049] Medical anticoagulant tube (containing EDTA-K2 anticoagulant) to collect fresh blood samples, centrifuge at 3000rpm for 10min, take the upper plasma into a new RNase-free centrifuge tube, centrifuge at 12000rpm for 10min at high speed to remove cell debris, and take the upper plasma (don’t The cell debris suck...

Embodiment 2

[0075] Example 2: MicroRNA-135a targets TRPM7 channel to inhibit hypoxia-induced myocardial fibrosis

[0076] Experimental animals: 1 to 3 days old suckling rats, male or female, provided by the Center of Comparative Medicine of Yangzhou University, animal certificate number: SCXK (苏)2016-0001.

[0077] Medicines and reagents: FBS (Gibico, USA), low-sugar DMEM (HyClone, USA), penicillin, streptomycin, Trypsin, EGTA (Biosharp), HEPEs, DEPC water, Trizol, BeyoECL Plus developer, RIPA lysis solution (strong) , BCA protein quantification kit (enhanced) (Biyuntian Bio), Cs-methanesulfonate (cesium methanesulfonate), 2-APB, Carvacrol (carvacrol), DMSO (Sigma Aldrich, United States), type II collagenase ( Worthington, USA), miRNA first strand cDNA synthesis (tailing method) kit, 5×SDS-PAGE loading buffer (Biotech), PrimeScript TM RTMaster Mix, Premix Ex Taq TM II (Dalianbao Bio), rabbit anti-TRPM7 protein primary antibody (Abcam, UK), rabbit anti-GAPDH internal control primary antibody...

Embodiment 3

[0147] Example 3: MicroRNA-135a targets TRPM7 channel to inhibit isoprenaline-induced myocardial fibrosis

[0148] Experimental animals: SPF grade SD rats, male, weighing 200-220g, purchased from the Comparative Medicine Center of Yangzhou University, certificate number: SCXK (Su) 2015-0002. Breed in an environment of 20±2℃, keep 12h light / 12h dark conditions, and freely feed in water. 1-3 days old suckling rats, male or female, purchased from the Center of Comparative Medicine, Yangzhou University, animal certificate number: SCXK (苏)2016-0002.

[0149] Medicines and reagents: I Reduced Serum Medium (Gibco, USA), lipo 6000 transfection reagent (Biyuntian Biotechnology), Isoprenaline hydrochloride (Sigma, USA), bovine serum albumin (Biosharp), the sources of other reagents and medicines are the same as in Example 2.

[0150] 1. Solution and culture medium

[0151] TRPM7 current extracellular fluid (mM): 140NaCl, 5KCl, 2CaCl 2, 20HEPEs, 10Glucose, NaOH adjusted to pH 7.4, stored at 4°...

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Abstract

The invention discloses application of miR-135a in the treatment of myocardial fibrosis, and belongs to the technical field of biomedicine. The anti-cardiac fibrosis mechanism of the miR-135a is explored as the research entry point, atrial fibrosis is induced by using atrial fibrillation, a model of cardiac fibrosis is induced by isoproterenol, culturing experiments of suckling mouse fibroblasts and 3T3 cells are combined, miR135a-TRPM7-TGF[beta]1/SMAD is interfered, a miR135a-TRPM7-NF-[kappa]B/MMP axis is used for investigating the role of the miR-135a in the myocardial fibrosis, new functions of the miR-135a are found to be used as drug targets to screen drugs for the prevention and/or treatment of the myocardial fibrosis and prepare drugs for the prevention and/or treatment of the myocardial fibrosis, and an effective new way for the treatment of the myocardial fibrosis is provided.

Description

Technical field [0001] The invention belongs to the field of biomedical technology, and specifically relates to the application of miR-135a in the preparation of medicines for treating myocardial fibrosis. Background technique [0002] Myocardial fibrosis is a difficult point in clinical treatment. It is an independent risk factor for cardiovascular disease morbidity and mortality. After the heart is damaged, cardiac fibroblasts in a resting state experience various stimuli and are awakened, leading to myocardial fibrosis. Myocardial fibrosis is manifested as an imbalance between the synthesis and degradation of extracellular matrix, the increase of collagen deposition in the interstitium, the imbalance of various types of collagen, and the disordered arrangement. If myocardial fibrosis is further aggravated, it is bound to damage the contractile function of the myocardium and induce heart failure. In addition, the abnormal electrical conduction of the myocardial tissue can lead...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61K45/06A61P9/00
CPCA61K31/7105A61K45/06A61P9/00
Inventor 汤依群刘勇慧吴艳许昊男刘婷婷马云天
Owner CHINA PHARM UNIV
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