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Method for detecting activity of spermidine/spermine N1-acetyltransferase

A technology of acetyltransferase and activity, applied in the field of detection of spermidine/spermine N1-acetyltransferase activity, which can solve problems such as inappropriate, rapid heart rate and slow action

Inactive Publication Date: 2012-02-08
上海拜瑞曼克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0045] 4. Pharmacology and Toxicology
[0049] ①The curative effect is better for mild and younger patients, but poor for severe and elderly patients;
[0050] ② It has better curative effect on muscle stiffness and dyskinesia, but poor curative effect on muscle tremor symptoms, such as long-term medication and larger doses can still be effective on the latter;
[0051] ③The effect is slow, and it usually takes 2 to 3 weeks to improve the objective signs, and the maximum curative effect is obtained after 1 to 6 months, but the effect is long-lasting and increases with the prolongation of the medication time
[0084] (3) Adverse reactions
[0096] ③ Monoamine oxidase inhibitors can prevent the degradation of dopamine in the body and enhance its effect, but they can lead to rapid heart rate and hypertensive crisis, so they should not be used together with this product
[0097] ④ It should not be used together with reserpine and adrenomimetic drugs
[0142] In conclusion, the analysis of existing technical data such as literature search shows that so far, no reports have been found on SSAT activity, enzyme kinetics, and acetylation assay methods of SSAT spermidine and spermine substrates.

Method used

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  • Method for detecting activity of spermidine/spermine N1-acetyltransferase
  • Method for detecting activity of spermidine/spermine N1-acetyltransferase
  • Method for detecting activity of spermidine/spermine N1-acetyltransferase

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Experimental program
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Embodiment 1

[0363] The liver cells of 20 rats (Wistar rats, female, 200-300g) were used, and the cells were obtained by Celsis in vitro technology and stored in liquid nitrogen. The day before use, the frozen hepatocytes were thawed in a 37°C water bath, suspended with the culture medium, centrifuged at 100×g for 10 minutes, and resuspended with Krebs-Hensleite buffer (KHB). Use 24-well plates for in vitro culture at 37°C with 5% CO 2 , 95% air, each hole is filled with about 2.5×10 5 Live liver cell stock solution 0.49ml, incubated for 10min. Then pre-incubation for 10min. In the liver cell solution, add L-dopa at 200 μM, inhibitor quercetin at 480 μM, add L-dopa at 200 μM, inhibitor methotrexate at 50 μM, and add L-dopa at 200 μM, inhibitor acetamine Phenol 2144μM, each substrate, inhibitor mixed solution 5μl to the culture plate, continue to incubate for 0, 30 and 60min. At the end of the incubation, 0.5 ml of ice methanol was added to each well to stop the reaction. Each group wa...

Embodiment 2

[0367] The liver cells of 20 rats (Wistar rats, female, 200-300 g) were used, and the cells were obtained by Celsis in vitro technology and stored in liquid nitrogen. The day before use, the frozen hepatocytes were thawed in a 37°C water bath, suspended with the culture medium, centrifuged at 100×g for 10 minutes, and resuspended with Krebs-Hensleite buffer (KHB). Use 24-well plates for in vitro culture at 37°C with 5% CO 2 , 95% air, each hole is filled with about 2.5×10 5 Live liver cell stock solution 0.49ml, incubated for 10min. Then pre-incubation for 10min. In the liver cell solution, add L-dopa at 20 μM, inhibitor quercetin at 480 μM, add L-dopa at 200 μM, inhibitor methotrexate at 50 μM, and add L-dopa at 200 μM, inhibitor acetamine Phenol 2144μM, each substrate, inhibitor mixed solution 5μl to the culture plate, continue to incubate for 0, 30 and 60min. At the end of the incubation, 0.5 ml of ice methanol was added to each well to stop the reaction. Each group wa...

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Abstract

The invention, relating to the technical field of medicine, relates to a method for detecting the activity of spermidine / spermine N1-acetyltransferase, that is, a quantitative technique of analyzing SSAT activity through detecting acetylated metabolites of the SSAT substrate. According to the invention, the N-acetylation is regulated mainly through or only through SSAT in SSAT pathway and the specific substrate in the N-acetylation regulation is L-dopa, partial metabolism of the substrate can generate an acetylated metabolite N-acetyl-L-dopa thought the introduction of SSAT, the SSAT upregulation is positively correlated with cancer, accordingly, L-dopa can be used as a cancer mark regulated by the SSAT upregulation, and a novel detection method is provided for early detection, early diagnosis, early treatment and the like of tumors. The method has the advantages of safely, efficiency, strong practicality, convenient and fast operating method, simple usage, high positive incidence, and remarkable effect, and can be used as an auxiliary means for preventing, diagnosing, detecting, protecting, treating and studying various tumors.

Description

technical field [0001] The present invention relates to the field of medical technology, in particular to a medical detection technology, more specifically to a spermidine / spermine N 1 -A method for acetyltransferase (abbreviation: SSAT) activity, and more specifically relates to a method for analyzing SSAT activity by detecting acetylated metabolites of SSAT substrates, wherein: the SSAT substrate is levodopa, levodopa Dopa is partially metabolized by induction of SSAT to produce the acetylated metabolite N-acetyl-L-dopa. Background technique [0002] (1) Research overview of levodopa [0003] 1 Overview [0004] 3-Hydroxy-L-tyrosine / L-β-(3,4-dihydroxyphenyl)alanine / L-3-(3,4-dihydroxyphenyl)alanine / L-(3,4 -Dihydroxyphenyl)alanine (Levodopa), also known as L-dopa (abbreviation: L-Dopa, L-DOPA), is the hydroxylation of tyrosine, which is the intermediate product of L-tyrosine synthesis of catecholamines in the body . As a drug, the common name is levodopa, also known as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
Inventor 郑启明阿曼得・拉希德李铁军周辉
Owner 上海拜瑞曼克生物科技有限公司
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