Method for detecting location of resveratrol in target cells

A technology of resveratrol and target cells, applied in the field of detection, can solve the problems of human radiation hazards, long detection cycle, high labeling costs, etc.

Active Publication Date: 2012-09-19
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Gester et al. used radionuclide (18F) or fluorescent probe-labeled resveratrol to detect its transport characteristics in the intestinal epithelium (Gester S, Wuest F, Pawelke B, Bergmann R , Pietzsch J. Synthesis and biodistribution of an 18F-labelled resveratrol derivative for small animal positron emission tomography. Amino Acids. 2005 Dec; 29(4):415-28), these methods can be used to detect The absorption of resveratrol in the intestinal tract and the characteristics of transmembrane transport, but the detection period is long, the labeling cost is high, there are human radiation hazards, and it is not easy to carry out in conventional laboratories

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  • Method for detecting location of resveratrol in target cells
  • Method for detecting location of resveratrol in target cells
  • Method for detecting location of resveratrol in target cells

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Example 1. Localization of Resveratrol in Human Umbilical Vein Endothelial Cells Detected by Laser Confocal Microscopy

[0033] Take human umbilical vein endothelial cells into a live cell culture dish, add resveratrol to the medium until the final concentration of resveratrol is 20 μM, and then incubate at a temperature of 37°C, 100% saturated humidity, and a volume fraction of 5%. CO 2 Treat in an incubator for 1 hour to obtain resveratrol-treated human umbilical vein endothelial cells; add the cell membrane red fluorescent probe (1,1'-dioctadecyl-3,3,3 ',3'-tetramethylindocarbocyanine perchlorate, Dil) (purchased from Beyond Biotechnology Co., Ltd.), dissolved in DMSO to a final concentration of 10 μM, and treated at 25°C for 5 minutes; the excitation wavelength was 322nm, and the reception wavelength To detect the fluorescent site of resveratrol under the condition of 397nm, the result is as follows image 3 shown. At the same time, the luminescent position of ...

Embodiment 2

[0037] Example 2. Detection of resveratrol localization in liver cells by laser confocal microscopy

[0038] Take the hepatic cells HepG2 and culture them in live cell culture dishes, add resveratrol to the medium until the final concentration of resveratrol is 1 μM, and then in the temperature of 37 ° C, 100% saturated humidity, volume fraction of 5% CO 2 Treat in an incubator for 1.5 hours to obtain resveratrol-treated hepatocytes; add cell membrane red fluorescent probe Dil (purchased from Beyond Biotechnology Co., Ltd.) to the obtained resveratrol-treated hepatocytes, and dissolve them with DMSO until The final concentration was 5 μM, and the temperature was 25°C for 20 minutes; the excitation wavelength was 322nm, and the reception wavelength was 350nm to detect the luminescence site of resveratrol, and the results were as follows: Figure 7 shown; at the same time, the luminescent position of Dil is detected under the condition that the excitation wavelength is 549nm ...

Embodiment 3

[0042] Example 3. Localization of resveratrol in breast cancer cells detected by confocal laser microscopy

[0043] Take the breast cancer cell MDA-MB-231 and culture it in a living cell culture dish, add resveratrol to the medium until the final concentration of resveratrol is 40 μM, and then in the temperature of 37 ° C, 100% saturated humidity, volume Fraction 5% CO 2 Treat in the incubator for 2 hours to obtain resveratrol-treated tumor cells; add the cell membrane red fluorescent probe Dil (purchased from Beyond Biotechnology Co., Ltd.) to the obtained resveratrol-treated breast cancer cells, and dissolve them with DMSO to The final concentration was 8 μM, and the temperature was 25°C for 10 minutes; the excitation wavelength was 322nm, and the reception wavelength was 500nm to detect the luminescence site of resveratrol, and the results were as follows: Figure 11 Shown; At the same time, when the excitation wavelength is 549nm and the emission wavelength is 565nm, th...

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Abstract

The invention discloses a method for detecting the location of resveratrol in target cells. Specifically, the method comprises the following steps of: firstly pretreating the target cells with resveratrol based on the autofluorescence property of resveratrol; then treating the target cells by using a specifically fluorescently-labeled subcellular organelle probe; and finally determining the distribution of resveratrol in the subcellular organelle of the target cells according to the overlap state of the autofluorescence of resveratrol and the fluorescence of the subcellular organelle probe in the cells. The method disclosed by the invention can be used for detecting the distribution of resveratrol in target cells, and has wide application prospect.

Description

[0001] technical field [0002] The invention relates to the field of detection, in particular to a method for detecting the localization of resveratrol in target cells. Background technique [0003] Resveratrol is a phytochemical widely present in many natural foods and medicines such as grapes, peanuts, Polygonum cuspidatum, Polygonum multiflorum, etc. Liver action. A variety of health foods or dietary supplements with resveratrol as the main ingredient have been launched in the United States, such as Longevity TM Anti-aging health food. The anti-tumor, anti-cardiovascular disease, and anti-senile dementia drugs developed with resveratrol as the lead compound have also been successfully screened, and some drugs have entered clinical trials and have good application prospects. However, the molecular mechanism of resveratrol's multiple biological effects has not been fully elucidated, and the intracellular targets are not clear. Proteins, etc.) are closely related to the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 易龙糜漫天陈明亮金鑫
Owner ARMY MEDICAL UNIV
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