CN104048961A - Detection method for mitochondria damages of nasopharynx mucosae cells - Google Patents
Detection method for mitochondria damages of nasopharynx mucosae cells Download PDFInfo
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- CN104048961A CN104048961A CN201410329563.5A CN201410329563A CN104048961A CN 104048961 A CN104048961 A CN 104048961A CN 201410329563 A CN201410329563 A CN 201410329563A CN 104048961 A CN104048961 A CN 104048961A
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Abstract
The invention provides a detection method for mitochondria damages of nasopharynx mucosae cells. The detection method comprises the following detection steps: taking nasopharynx seepage; putting the nasopharynx seepage into a sampling bottle which is prefilled with a buffering solution and uniformly mixing; adding 1ml of a reagent A and 1ml of a reagent B into the sample bottle in sequence; uniformly mixing and observing the color change within two minutes; and comparing a detection result with a colorimetric plate. The detection method is simple and rapid and can be used for detecting the result that the concentration of the lowest free ferrous protoporphyrin can be up to 5ng/ml; the sensitivity is very high.
Description
Technical field
The present invention relates to a kind of detection method, be specifically related to a kind of detection method of pharynx nasalis mucomembranous cell injury of mitochondria.
Background technology
Mitochondria is most important organelle in human body cell, is " the power factory " of cell.To impaired pharynx nasalis mucomembranous cell, mitochondrial detection comprises detection, 2mtDNA damage check, intracellular calcium concentration detection of detection, the film potential of detection to Mitochondrial permeability transition pore (mitochon2dria permeability transition pore, MPTP), membrane phospholipid etc.It is very complicated that this type of detects operation, needs accurate instrument and equipment, and common laboratory has been difficult to, and pharynx nasalis mucomembranous cell injury of mitochondria detected and be extremely restricted.Before as above highly difficult detection, use a kind of method simple to operate, with low cost, whether pharynx nasalis mucomembranous cell mitochondria is damaged and carry out preliminary examination, can whether damage the enlightenment effect of playing to pharynx nasalis mucomembranous cell mitochondria, play indicative function to whether needing further to carry out as above highly difficult detection.Therefore, a kind of pharynx nasalis mucomembranous cell injury of mitochondria detection technique tool of research invention has very important significance.
Summary of the invention
For the problem that solves complicated operation in the method that detects pharynx nasalis mucomembranous cell injury of mitochondria in prior art, apparatus expensive, is difficult to carry out, the invention provides and a kind ofly detect pharynx nasalis sepage cell free ferroprotoporphyrin and then understand the detection method that pharynx nasalis mucomembranous cell mitochondria whether damages, method provided by the invention is simple, with low cost, examination pharynx nasalis mucomembranous cell mitochondria whether damages rapidly.
The detection method of pharynx nasalis mucomembranous cell injury of mitochondria provided by the invention, comprises following detecting step:
1) get pharynx nasalis sepage 0.5-1 ml;
2) pharynx nasalis sepage is placed in to the sampling jar that is preinstalled with damping fluid, mixes; In specimen bottle, be preinstalled with acetate buffer solution, the amount of damping fluid is 2ml, mixes, and obtains sample liquid; Preferably, acetate buffer solution pH value is 4.5;
3) in specimen bottle, order adds reagent A 1ml, reagent B 1ml; Mix, in 2 minutes, observe change color, with color board Determination result;
Described reagent A is two (4. dimethylamino phenyl) methane, after dissolving, is mixed with solution with methyl alcohol; Preferably, two (4. dimethylamino phenyl) methane solution concentration is 0.002mol/L;
Described reagent B is superoxol; Preferably, superoxol is 1.5%.
Aobvious blueness or mazarine, can judge in sample and contain free ferroprotoporphyrin, mitochondria has damage.
Under the effect of some factor,, there is energetic supersession reprogrammed in the imbalance of pharynx nasalis mucomembranous cell stable state.There is Wa Bogeshi effect in the metabolism of pharynx nasalis mucomembranous cell, cause hypoxemia and reduction-state microenvironment in cell, excited oxygen receptor and Anoxia-Signaling Pathway path, impel anoxic idiosyncratic transcription factor-hypoxia inducible factor and second messenger-active chalcogen activity to increase, the polarity value that changes the cell proteins (HP) such as cytochrome, the cell free ferroprotoporphyrin (Cells Free Ferrous Protoporphyrin) that makes to occupy in albumen hydrophobic core is separated out.Cell free ferroprotoporphyrin expedites the emergence of multiple free radical, causes the oxidative damage of Cell membrane lipids, lipoprotein, cytoskeleton, DNA etc., can make mitochondria occur mitochondria unit price electronics seepage (univalent leak); Ca2+ overload in born of the same parents, triggers mitochondria picked-up Ca2+ and makes Ca2+ in mitochondria inner accumulated; Suppress the function of dehydrogenasa in mitochondria; More than change and can make mitochondrial swelling, ridge fracture disintegration, calcium deposition, outer membrane rupture and matrix excessive.The cell free ferroprotoporphyrin amount of separating out is proportionate with pharynx nasalis mucomembranous cell injury of mitochondria degree.Cell free ferroprotoporphyrin after separating out, can in pharynx nasalis sepage, survey and.As examination means, measure cell free ferroprotoporphyrin content in pharynx nasalis sepage, can point out pharynx nasalis mucomembranous cell mitochondria whether to damage.
Detection method provided by the invention, simple and practical, with low cost, examination fast go out in pharynx nasalis mucomembranous cell containing the sample of free ferroprotoporphyrin, containing free ferroprotoporphyrin sample and may contain free ferroprotoporphyrin sample, and then understand pharynx nasalis mucomembranous cell mitochondria and whether damage, for further checking and prepare; The concentration of the detectable minimum free ferroprotoporphyrin of the present invention can reach 5ng/ml, highly sensitive.
Embodiment
Below in conjunction with embodiment, the present invention is described further.
Embodiment 1
Detect by the following method sample:
1) get pharynx nasalis sepage 0.5-1 ml;
2) pharynx nasalis sepage is placed in to the sampling jar that is preinstalled with damping fluid, mixes; In specimen bottle, being preinstalled with pH value is 4.5 acetate buffer solutions, and the amount of damping fluid is 2ml, mixes, and obtains sample liquid;
3) in specimen bottle, order adds reagent A 1ml, reagent B 1ml; Mix, in 2 minutes, observe change color, with color board Determination result;
Described reagent A is that concentration is 0.002mol/L bis-(4. dimethylamino phenyl) methane, after dissolving, is mixed with solution with methyl alcohol;
Described reagent B is 1.5% superoxol.
[criterion]
In specimen bottle, liquid does not develop the color or micro-blueness () feminine gender
Blue (+) positive
Mazarine (++) strong positive
experimental data:
Get respectively and detect as not developing the color, shows blueness and each three examples of navy blue sample, carry out the detection of cell free ferroprotoporphyrin with mass spectrometer, testing result is as follows: not survey and cell free ferroprotoporphyrin do not develop the color all; Blueness is 5 ng/ml, 22ng/ml, 131ng/ml; Mazarine is 207ng/ml, 0.12 μ g/ml, 0.37 μ g/ml.
Can find out by above data, sample to cell free ferroprotoporphyrin content more than 5ng/ml, this screening method just can be preliminary judgement, content this detection method of sepage more than 20ng/L just can be defined as containing cell free ferroprotoporphyrin in the sepage of pharynx nasalis cell, prompting pharynx nasalis mucomembranous cell injury of mitochondria, so this screening method is practical.
Claims (4)
1. a detection method for pharynx nasalis mucomembranous cell injury of mitochondria, comprises following detecting step:
1) get pharynx nasalis sepage 0.5-1 ml;
2) pharynx nasalis sepage is placed in to the sampling jar that is preinstalled with damping fluid, mixes; The damping fluid of prepackage is acetate buffer solution, and the amount of damping fluid is 2ml, mixes, and obtains sample liquid;
3) in specimen bottle, order adds reagent A 1ml, reagent B 1ml; Mix, in 2 minutes, observe change color, with color board Determination result;
Described reagent A is two (4. dimethylamino phenyl) methane, after dissolving, is mixed with solution with methyl alcohol;
Described reagent B is superoxol.
2. detection method according to claim 1, is characterized in that, described acetate buffer solution pH value is 4.5.
3. detection method according to claim 1, is characterized in that described two (4. dimethylamino phenyl) methane solution concentration is 0.002mol/L.
4. detection method according to claim 1, is characterized in that, the simmer down to 1.5% of described superoxol.
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Cited By (3)
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| RU2619873C1 (en) * | 2016-03-22 | 2017-05-18 | Федеральное бюджетное учреждение науки "Федеральный научный центр медико-профилактических технологий управления рисками здоровью населения" (ФБУН "ФНЦ медико-профилактических технологий управления рисками здоровью населения") | Method for diagnostics of nasopharynx chronic inflammatory diseases associated with inhalation exposure to benzene and formaldehydein for children of 4-12 years |
| CN107132217A (en) * | 2017-05-11 | 2017-09-05 | 山东天倪生物技术有限公司 | Cervical cell warburg effect simple detection reagent and preparation method and application thereof |
| RU2763821C1 (en) * | 2021-03-23 | 2022-01-11 | Дмитрий Валерьевич Трусов | Method for early diagnosis of chronic diseases of the paranasal sinuses and palatine tonsils |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2619873C1 (en) * | 2016-03-22 | 2017-05-18 | Федеральное бюджетное учреждение науки "Федеральный научный центр медико-профилактических технологий управления рисками здоровью населения" (ФБУН "ФНЦ медико-профилактических технологий управления рисками здоровью населения") | Method for diagnostics of nasopharynx chronic inflammatory diseases associated with inhalation exposure to benzene and formaldehydein for children of 4-12 years |
| CN107132217A (en) * | 2017-05-11 | 2017-09-05 | 山东天倪生物技术有限公司 | Cervical cell warburg effect simple detection reagent and preparation method and application thereof |
| CN107132217B (en) * | 2017-05-11 | 2019-11-08 | 山东天倪生物技术有限公司 | Cervical cell warburg effect simple detection reagent and preparation method and application thereof |
| RU2763821C1 (en) * | 2021-03-23 | 2022-01-11 | Дмитрий Валерьевич Трусов | Method for early diagnosis of chronic diseases of the paranasal sinuses and palatine tonsils |
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Application publication date: 20140917 |