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Application of hypoxia-inducible factor -2alpha as action target in prevention/treatment of Parkinson's disease

A hypoxia-inducible factor, Parkinson's disease technology, applied in the field of neurobiological cells, can solve the unreported regulation mechanism and specific target of iron metabolism in astrocytes, and the inability to determine the source of iron in astrocytes. Regulatory mechanism of iron metabolism in glial cells, lack of role of astrocytes, etc.

Pending Publication Date: 2020-02-11
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the research on the interaction between astrocytes and DA neurons mainly focuses on the protective effect of astrocytes on DA neurons, such as the neuroprotective effect of BDNF and GDNF, and the anti-oxidative stress of SOD However, there are few studies on the mechanism of iron metabolism in astrocytes. There is no report on the regulation mechanism and specific targets of iron metabolism in astrocytes, and there is no astrocytes as the support of neurons. The role of cells in iron deposition in PD neurons, the source of excess iron in neurons and the regulatory mechanism of iron metabolism in astrocytes cannot be determined

Method used

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  • Application of hypoxia-inducible factor -2alpha as action target in prevention/treatment of Parkinson's disease
  • Application of hypoxia-inducible factor -2alpha as action target in prevention/treatment of Parkinson's disease
  • Application of hypoxia-inducible factor -2alpha as action target in prevention/treatment of Parkinson's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Primary culture of astrocytes

[0039] 1.1 Experimental animals

[0040] Newborn 24h Wistar suckling mice (provided by the Animal Center of Qingdao Institute of Drug Control) were selected as experimental animals for the primary culture of astrocytes.

[0041] 1.2 Experimental reagents

[0042] DMEM / F-12(1:1): Gibco product;

[0043] Fetal bovine serum (FBS Gold): Gibco company product, EU grade;

[0044] Polylysine, boric acid, sodium tetraborate: products of Sigma;

[0045] Penicillin-streptomycin solution (100X), trypsin (Trypsin): Products of Jiangsu Haimen Beyontian Institute of Biotechnology.

[0046] 1.3 Experimental Instruments

[0047] Ultra-clean workbench: Suzhou Purification Equipment Co., Ltd.;

[0048] CO 2 Incubator: American Thermo Fisher Scientific products;

[0049] Ordinary centrifuge: Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.;

[0050] Thermostat and oven: Germany MEMMERF products;

[0051] -20°C and 4°C refrigerators...

Embodiment 2

[0073] Example 2 Primary Culture of Ventral Mesencephalon (VM) Neurons

[0074] 2.1 Experimental animals

[0075] As experimental animals, 14-day pregnant Wistar mice (provided by the Animal Center of Qingdao Institute of Drug Control) were selected, placed at a room temperature of 20° C., and lived under the conditions of 12:12 hours of day and night cycle light, free access to food and water.

[0076] 2.2 Experimental reagents

[0077] DMEM / F-12(1:1): Gibco product;

[0078] B27: Gibco products;

[0079] Fetal bovine serum (FBS Gold): Gibco company product, EU grade;

[0080] Polylysine, boric acid, sodium tetraborate: products of Sigma;

[0081] Penicillin-streptomycin solution (100X), trypsin (Trypsin): Products of Jiangsu Haimen Beyontian Institute of Biotechnology.

[0082] 2.3 Experimental Instruments

[0083] Ultra-clean workbench: Suzhou Purification Equipment Co., Ltd.;

[0084] CO 2 Incubator: American Thermo Fisher Scientific products;

[0085] Ordinary cent...

Embodiment 3

[0105] Example 3 Western blot detection of activation of HIF-1α and HIF-2α and protein expression of DMT1 and FPN1

[0106] 3.1 Experimental reagents

[0107] 6-hydroxydopamine (6-hydroxydopamine, 6-OHDA), ascorbic acid: products of Sigma Company.

[0108] HIF-2α translation inhibitor: Millipore product

[0109] Bay87-2243: Product of Selleck, HIF-1α-specific inhibitor

[0110] Acrylamide, phenylmethylsulfonamide (PMSF): product of Genview Company.

[0111] Ammonium persulfate (APS), Tween-20 (Tween 20), sodium dodecyl sulfate (SDS, electrophoresis grade), NonidetP40, pepstatin, aprotinin, leupeptin, deoxycholic acid Sodium (Nadeoxycholate), rabbit anti-β-actin antibody, HRP-labeled goatanti-rabbit IgG: products of Sigma.

[0112] N', N'-methylenebisacrylamide: product of Bebco Company.

[0113] Glycine (Glycine, Glycine, electrophoresis grade), trishydroxymethylaminomethane (Tris), Tris·Base, Tris·HCl, ethylenediaminetetraacetic acid (EDTA): products of Solarbio.

[011...

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Abstract

The invention, which belongs to the field of neurobiological cells, provides application of hypoxia-inducible factor -2alpha as an action target in prevention / treatment of the Parkinson's disease. According to the invention, the action of an astrocyte hypoxia inducible factor HIF-2alpha in intracerebral iron metabolism regulation and a regulation mechanism are studied for the first time and different iron metabolism modes of astrocytes and neurons are clarified, so that a source of nerve cell high-iron is found for PD iron deposition and the PD iron deposition is caused by acceleration of astrocyte iron transport specifically. However, the core element for promoting the iron transport acceleration of the astrocyte is HIF-2alpha. Therefore, a new action target HIF-2alpha is provided for preventing / treating PD iron deposition and novel research thought and experimental evidences are provided for the iron transport mechanism of high-iron-derived astrocytes of nerve cells. The applicationof hypoxia-inducible factor -2alpha as an action target in prevention / treatment of the Parkinson's disease has the potential immeasurable social and economic values for research and development of drugs for preventing / treating PD.

Description

technical field [0001] The invention belongs to the field of neurobiological cells, and in particular relates to the application of hypoxia-inducible factor-2α as an action target in the prevention / treatment of Parkinson's disease. Background technique [0002] Parkinson's disease (PD) is a degenerative disease of the central nervous system that mainly occurs in middle-aged and elderly people. Loss of dopamine (DA) neurons in the substantia nigra compacta. At present, the research on iron deposition in PD mainly focuses on DA neurons, but in fact, a variety of glial cells in the brain also play an important role in the regulation of iron homeostasis, especially astrocytes, which account for about 10% of the central nervous system (central nervous system). Nervous system, CNS) cells account for 90% of the total number of cells, which can absorb, store and release iron, precisely regulate brain iron transport and brain iron content, and play an important role in the regulatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/6875G01N33/56966G01N2500/00G01N2333/70567G01N2800/2835A61K31/137A61P25/16C12N5/0622C12N2509/00C12N2502/081A61K35/30
Inventor 谢俊霞王俊许曼曼
Owner QINGDAO UNIV
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