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Detection method of heavy metal copper cytotoxicity based on nitric oxide synthase nos

A technology of nitric oxide and cytotoxicity, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, and can solve the problems of incompletely clear molecular mechanisms

Active Publication Date: 2018-01-30
SHANGHAI JIAO TONG UNIV
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  • Application Information

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Problems solved by technology

However, the specific toxicity mechanism of heavy metal copper, especially the molecular mechanism related to signal transduction, is still not fully understood.

Method used

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  • Detection method of heavy metal copper cytotoxicity based on nitric oxide synthase nos
  • Detection method of heavy metal copper cytotoxicity based on nitric oxide synthase nos
  • Detection method of heavy metal copper cytotoxicity based on nitric oxide synthase nos

Examples

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Embodiment 1

[0036] The test object of this embodiment: copper chloride.

[0037] Preparation method: Dissolve the standard product in sterile water to prepare a 0.1mol / L mother solution and store it at -20°C. Dilute it with culture medium to the required concentration before operation.

[0038] Model organism: human breast cancer MCF‐7 cells.

[0039] Reagents: RNA Rapid Extraction Kit (Spin-column was purchased from Bioteke; N-(6-methoxy-8-quinolyl) p-toluenesulfonamide (TSQ) was purchased from AAT Bioquest; N-acetyl-L-cysteine Amino acid (LNAC), L‐arginine (L‐Arg), dihydrorhodamine 123 (DHR), eNOS antibodies were purchased from Sigma; nNOS,, iNOS, β‐actin antibodies were purchased from Abcam; p‐elF2α Antibodies were purchased from Invitrogen; Western luminescence kits were purchased from Pierce; Fluo-3 / AM and nitric oxide detection kits were purchased from Beyond Biotechnology Research Institute; reverse transcription kits and RT-PCR kits were purchased from Bao Bioengineering Dalian Co...

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Abstract

A method for detecting cytotoxicity of heavy metal copper based on nitric oxide synthase NOS in the technical field of heavy metal toxicity, using heavy metal copper to depolymerize the NOS dimer by replacing zinc ions and calcium ions in the NOS dimer, NOS dimer The amount of synthesis decreased, and the expression of NOS mRNA was up-regulated in a feedback manner; on the other hand, the depolymerization of NOS dimers was accompanied by the increase of the number of NOS monomers, the increase of active oxygen content in cells, the phosphorylation of cell translation initiation factors, and the protein Synthesis stops, and eventually leads to the toxic mechanism of apoptosis. Nitric oxide synthase NOS is used for the detection of heavy metal copper cytotoxicity, which provides a new solution for the biosafety assessment of heavy metal pollution and the treatment of heavy metal poisoning and the prevention and treatment of related diseases. Program.

Description

technical field [0001] The invention relates to a method in the technical field of heavy metal toxicity, in particular to a method for detecting heavy metal copper cytotoxicity mediated by nitric oxide synthase NOS signaling pathway. Background technique [0002] Heavy metal copper is an essential trace element in living organisms, a cofactor of various enzymes, and participates in electron transfer in biological systems. At the same time, studies have shown that after exposure to copper ions, a large amount of reactive oxygen species are produced, leading to apoptosis of breast cancer epithelial cells. In neurons, copper exposure-induced apoptosis is associated with increased intracellular NO synthesis. Nitric oxide synthase (Nitric Oxide Synthase, NOS) is a kind of isoenzyme, which is the only kind of specificity in organisms that uses L‐Arginine (L‐Arginine) as a substrate and uses oxygen to generate NO Nitrogen (NO) enzymes. NO is a soluble gas. As a signal molecule, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851C12Q1/26C12Q1/02
Inventor 周培陆伟钟玲盈
Owner SHANGHAI JIAO TONG UNIV
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