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A method for evaluating proliferation toxicity of cigarette smoke based on bromodeoxyuridine incorporation

A technology of bromodeoxyuridine and cigarette smoke, which is applied in the field of determination of cytotoxicity outside of cigarette smoke, can solve the problems of less research and achieve the effects of quick and easy operation, improved detection sensitivity, and high sensitivity

Active Publication Date: 2015-11-18
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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Problems solved by technology

[0004] At present, the methods used in the evaluation of the cytotoxicity of cigarette smoke are mostly based on the principle of cell survival rate measurement, and there are few studies on the effect of cigarette smoke exposure on cell proliferation and toxicity. Cell proliferation toxicity evaluation method, quantitative and accurate evaluation of cigarette smoke particulate matter and cytotoxicity

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  • A method for evaluating proliferation toxicity of cigarette smoke based on bromodeoxyuridine incorporation
  • A method for evaluating proliferation toxicity of cigarette smoke based on bromodeoxyuridine incorporation
  • A method for evaluating proliferation toxicity of cigarette smoke based on bromodeoxyuridine incorporation

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Embodiment Construction

[0050] The present invention will be further described below in conjunction with the examples, but the present invention is not limited.

[0051] In order to investigate the cell proliferation toxicity of cigarette sample 2 smoke particulate matter on A549 cells, the following experiments were carried out:

[0052] 1. Solution preparation

[0053] (1) Cigarette smoke exposure solution:

[0054] Cigarette samples were smoked using a rotary 20H smoking machine (Germany, Borgwaldt Company). The smoking mode was in accordance with ISO requirements, and the particulate phase components of cigarettes were captured by Cambridge filters. A total of 182 mg of smoke particulate matter was captured by smoking 20 cigarettes. After taking out the Cambridge filter, add 18.2 mL of dimethyl sulfoxide (DMSO) and shake for half an hour to obtain a smoke with a final concentration of 10 mg / mL. Particulate phase extract. Dilute with cell culture medium to prepare a series of smoke exposure sol...

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Abstract

The invention relates to a cigarette smoke cell proliferation toxicity evaluation method based on bromodeoxyuridine (BrdU) doping. The method is characterized by comprising the following steps: (1) preparing a solution; (2) carrying out cell inoculation; (3) contaminating cigarette smoke and doping BrdU; (4) assaying the doped amount of BrdU; (5) resulting and analyzing. The method has the following characteristics that through preparing a smoke granule-phase matter extracting solution and a smoke gas-phase matter absorbing solution, the cell toxicity of granule-phase and gas-phase matters of smoke can be inspected respectively; by a cell applicability validation step, the method can be applicable to a variety of adherent culture cells and can be used for inspecting the cell proliferation toxicity to different cell lines caused by cigarette smoke; the sensitivity of detection is improved through optimizing the doping time of the BrdU and the concentration of cell membrane permeation liquid; the best contaminating concentration is determined through optimizing the contaminating concentration of the cigarette smoke, so as to obtain the optimal dose-effect curve. In addition, the method further has the advantages of quickness, simplicity and convenience in operation, high sensitivity and stable results.

Description

technical field [0001] The invention relates to the measurement of extragas cytotoxicity of cigarette smoke, in particular to a quantitative measurement method for analyzing cell proliferation toxicity of cigarette smoke particulate matter or gas phase matter after exposure based on the principle of bromodeoxyuridine (BrdU) incorporation. Background technique [0002] Carrying out in vitro toxicology experiments is an important method to evaluate the health hazards of smoking. Among them, cytotoxicity plays an important role in many pathological processes including cancer and inflammation, and cytotoxicity test is an important way to understand the mechanism of interaction between compounds and cells and tissues. How to scientifically and accurately evaluate the harmfulness of cigarette products and tobacco additives, currently there is no uniform method and procedure in the world. Detection of cytotoxicity can be performed by various biological endpoints, such as counting ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
CPCG01N21/31G01N33/53
Inventor 陈欢刘彤韩书磊吴帅宾付立伟张小涛石龙凯侯宏卫胡清源
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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