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Preparation method of oxidized low-density lipoprotein and application technology thereof

A low-density lipoprotein, oxidized technology, applied in the preparation of test samples, material inspection products, peptide sources, etc.

Inactive Publication Date: 2010-07-28
XUZHOU CARDIOVASCULAR INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cucina A et al. found that oxidized low-density lipoprotein (ox-LDL) can stimulate smooth muscle cells to secrete bFGF and endothelial cells to secrete TGF-β1, suggesting that ox-LDL may play an important role in the process of myocardial fibrosis, especially in the re-recognized Intensive lipid-lowering therapy and blocking the signaling pathway of ox-LDL can be used as one of the measures to prevent and treat myocardial fibrosis, which has practical significance for clinical treatment and is currently in the research stage at home and abroad

Method used

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  • Preparation method of oxidized low-density lipoprotein and application technology thereof
  • Preparation method of oxidized low-density lipoprotein and application technology thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Extraction and identification of oxidized low-density lipoprotein:

[0011] Normal human serum was extracted and centrifuged by a high-speed centrifuge to remove the upper layer of chylomicrons to adjust the density. The ultracentrifuge was centrifuged twice, and the upper layer of LDL was taken, identified by agarose gel electrophoresis, and then ox-LDL was obtained by catalytic oxidation with copper sulfate. The oxidation value was measured by agarose gel electrophoresis and MDA kit, and the protein concentration was determined by BCA method.

Embodiment 2

[0013] Study on the promotion of myocardial fibrosis by ox-LDL in vitro:

[0014] 1. Liquid preparation

[0015] ①1mM KH solution containing calcium (mM·L -1 ): NaCl 119, KCl 4.7, MgSO 4 ·7H 2 O 0.94, KH 2 PO 4 1.2, NaHCO 3 25, Glucose H 2 O 11.5, CaCl 2 1. Pass 5% CO 2 +95%O 2 Mixed gas, pH 7.4. ② Calcium-free solution (mM·L -1 ): NaCl 120, KCl 5.4, MgSO 4 ·7H 2 O 5, Sodium pyruvate 5, Glucose H 2 O 20, Taurine 20, HEPES 10. ③Add 5mM NTA and 12μM-14μM CaCl to the calcium-free solution before separation 2 It is low calcium solution, adjust pH=6.95, fill with 100% O 2 . ④Enzyme solution, add 1mg·mL on the basis of calcium-free solution -1 Collagenase II and 50 μM CaCl 2 , after dissolving, adjust pH=7.4, fill with 100% O 2 . ⑤ M199 culture medium, add the following additives to the M199 medium (mM·L -1 ): Ascorbic Acid 100, Creatine 5, Taurine 5, Carnitine 2, Insuline 0.0001 and 0.2% BSA. Routinely add 100UPenicillin-100mg Streptomycin.

[0016] 2. Isola...

Embodiment 3

[0035] ox-LDL promotes myocardial fibrosis in vivo:

[0036] 1. Extraction and identification of ox-LDL

[0037] Normal human serum was extracted, centrifuged in a high-speed centrifuge to remove the upper layer of chylomicrons and then adjusted for density, centrifuged twice in an ultracentrifuge, and the upper layer of LDL was collected, identified by agarose gel electrophoresis, and then ox-LDL was obtained by copper sulfate catalytic oxidation. The oxidation value was measured by agarose gel electrophoresis and MDA kit, and the protein concentration was determined by BCA method.

[0038] 2. Experimental grouping Experimental grouping Blank control group, LDL control group and experimental group (8 rats in each group).

[0039] 3. Method of administration Tail vein injection.

[0040] PBS buffer, LDL and ox-LDL were injected into the blood vessels of the rats through the tail vein, once a day. The blank control group was injected with the corresponding volume of PBS buff...

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Abstract

The invention relates to a preparation method of low-density lipoprotein, in particular to a preparation method of oxidized low-density lipoprotein and application thereof, belonging to the technical field of the medical test. Through extraction and identification of the oxidized low-density lipoprotein and research on promoting myocardial fibrosis of the oxidized low-density lipoprotein in vitro, the method utilizes the effect of the extracted oxidized low-density lipoprotein on the myocardial fibrosis to investigate the occurrence mechanism of the myocardial fibrosis and can promote the research and development of the drug against the myocardial fibrosis through blocking the signal pathway.

Description

technical field [0001] The invention relates to a preparation method of low-density lipoprotein, in particular to a preparation method and application of oxidized low-density lipoprotein, and belongs to the technical field of medical detection. Background technique [0002] Myocardial fibrosis refers to the excessive accumulation of collagen fibers in the myocardial extracellular matrix (ECM), a significant increase in collagen concentration, or changes in collagen composition. It is the result of the imbalance of collagen synthesis and degradation metabolism. This pathological change exists in many cardiovascular diseases such as hypertrophic cardiomyopathy and heart failure. It is now believed to be closely related to arrhythmia, cardiac dysfunction and even sudden cardiac death. . Myocardial fibrosis is a complex pathological process, the main causes of which are long-term pressure overload, inflammatory response, apoptosis, and oxidative stress, involving neuroendocrine...

Claims

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Application Information

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IPC IPC(8): C07K14/47G01N33/53G01N1/28G01N1/30G01N1/42
Inventor 宫海滨徐锁平苏景力王洁庞慧李春梅
Owner XUZHOU CARDIOVASCULAR INST
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