CN114894910B - New application of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine detection reagent - Google Patents
New application of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine detection reagent Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention relates to the field of in vitro diagnostic reagents, and relates to a novel application of a 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine detection reagent. The invention discovers for the first time that the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in the peripheral blood plasma of patients suffering from depression is significantly higher than that of people without depression. The reagent for detecting the 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is used for preparing the depression diagnosis kit, so that the effective diagnosis of depression can be realized.
Description
Technical Field
The invention relates to the field of in vitro diagnostic reagents, in particular to application of a novel application of a 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine detection reagent.
Background
Depression is now the most common psychological disorder, with continuous long-term mood depression as the major clinical feature, the most important type of modern human mental disorder. Clinically, the mood is low and the reality is too careless, the mood is low and subsides for a long time, from the beginning of smoldering to the last of sadness, spewing, pain, pessimistic and aversion, the feeling of living is hopefully afflicting itself every day, negatively, evading, and finally, the feeling of suicidal tendency and behavior is even more. At the same time, patients also suffer from somatization symptoms such as chest distress and shortness of breath.
For the diagnosis of depression, the internationally common diagnostic criteria are typically ICD-10 and DSM-IV. ICD-10 is mainly adopted in China. In addition, diagnosis of depression by a biomarker is also a common method, for example, "application of CN201811553503.6D-serine as a marker in preparation of a depression diagnosis kit and depression diagnosis kit" propose diagnosis of depression by using D-serine as a marker; "CN201911236135.7 is used for distinguishing biomarker of depression and non-depression and diagnostic kit comprising the same", and the specific sequence polypeptide is used as a marker for diagnosing depression. However, to improve the accuracy of diagnosis, and to meet the needs of various complex clinical situations, it is desirable to be able to further develop more diagnostic agents for depression.
The structural formula of the 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is as follows:
1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine contains polyunsaturated fatty acids and plays a role in normal growth and development. The compound is easily oxidized, resulting in various fragmented and non-fragmented products. These products have the functions of regulating intracellular signal transduction, inducing apoptosis, stimulating ROS production, promoting or inhibiting inflammation, etc.
However, the connection between 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine and depression has not been reported at present.
Disclosure of Invention
The invention aims to provide a novel application of a 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine detection reagent and an application of the detection reagent of the compound in preparation of a depression diagnosis kit.
The technical scheme of the invention comprises the following steps:
use of a reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in the preparation of a depression diagnostic kit.
Preferably, the reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is a reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma.
Preferably, the reagent is a liquid chromatography-mass spectrometry detection reagent.
Preferably, the liquid chromatography-mass spectrometry combined detection reagent comprises 1-heptadecanoyl-glycerol-3-phosphorylcholine, glycocholic acid- (glycine-1-13C) -hydrate, methanol and acetonitrile.
The invention also provides a depression diagnosis kit which comprises a reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine.
Preferably, the reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is a reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma.
Preferably, the reagent is a liquid chromatography-mass spectrometry detection reagent.
Preferably, the liquid chromatography-mass spectrometry combined detection reagent comprises dissolved 1-heptadecanoyl-glycerol-3-phosphorylcholine, glycocholic acid- (glycine-1-13C) -hydrate, methanol and acetonitrile.
The invention discloses a 1-heptadecanoyl-glycerol-3-phosphorylcholine which is used as an internal reference compound for liquid chromatography-mass spectrometry combination detection, and the English name of the internal reference compound is that
1-heptadecanyl-2-hydroxy-sn-glycero-3-phosphonine (LPC 17:0) having the following structural formula:
the Glycocholic acid- (glycine-1-13C) -hydrate is used as an internal reference compound for liquid chromatography-mass spectrometry combination detection, and has the following structural formula:
the key point of the invention is that the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma is determined to be obviously related to the risk of suffering from depression, so that the risk of suffering from depression can be judged by detecting the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma, and as means for specifically detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma, various means disclosed in the prior art can be adopted.
The invention provides a novel depression diagnosis marker and a novel depression diagnosis kit, which can realize effective diagnosis of depression; and Zhou Xiejiang outside the human body can be used as a detection sample, so that the damage to the patient is low. The invention has good application prospect.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to the following embodiments. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
Figure 1 is a graph of the working curve (ROC) of subjects with respect to the concentration of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine associated with depression in example 1.
Detailed Description
In the following examples, reagents and materials not specifically described are commercially available.
EXAMPLE 1 relation of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine to depression in plasma
The liquid chromatography mass spectrometry used in this experiment analyzed the outer Zhou Xiejiang-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine concentration.
The experimental method comprises the following steps: the liquid chromatography mass spectrometry system UPLC H class & Quadranipole TQ-XS (Waters, milford, mass., USA), massLynx V4.2 software was used for instrument control, data acquisition and analysis.
The experimental steps are as follows:
1. 40. Mu.L of external Zhou Xiejiang was taken, 10. Mu.L of internal standard (1-heptadecanoyl-glycerol-3-phosphorylcholine: 1. Mu.g/mL; glycocholic acid- (glycine-1-13C) -hydrate: 50. Mu.g/mL) was added, and 150. Mu.L of methanol-acetonitrile solution (V/V=1:1) was added;
2. vortex for 30s and stand at-20 ℃ for 20min. Centrifuging at 14000rpm at 4deg.C for 10min;
3. 150 μl of the supernatant was loaded into an LC-MS sample vial with liner for analysis
4. Chromatographic conditions: accQ-Tag Ultra chromatography column (2.1 mm. Times.100 mm,1.7 μm) from Waters, the column temperature was maintained at 45℃and the sample volume was 3. Mu.L.
5. Mass spectrometer operating parameters: ionspray voltage, (+ -.) 4500V; source temperature,550 ℃; curtaingas, 20; CAD gas,8; nebulizer gas (GS 1), 50; auxiliary gas (GS 2), 50; EP,10; CXP,10. Electrospray ion source is in positive and negative double ion source mode, MRM condition and standard curve detection are shown in the following table.
The test results of healthy persons (n=49) and depressed patients (n=42) were compared, and the average concentration of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in the peripheral blood of healthy persons was 9125.048571ng/mL, and the average concentration of Zhou Xiejiang-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in depressed patients was 12235.7888ng/mL. Experimental results prove that the concentration of Zhou Xiejiang-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is obviously increased outside a patient suffering from depression, and a subject work (ROC) curve is shown in figure 1, and the result shows that the area under the curve reaches 0.9519, which indicates that the compound has the identification capability on depression and has high diagnostic value on depression.
Example 2A kit for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in plasma
The kit of this example contains the following reagents: 1-heptadecanoyl-glycerol-3-phosphorylcholine, glycocholic acid- (glycine-1-13C) -hydrate, methanol and acetonitrile.
Wherein, the 1-heptadecanoyl-glycerol-3-phosphorylcholine and glycocholic acid- (glycine-1-13C) -hydrate can be packaged independently or prepared into solution package (the concentration is 1-heptadecanoyl-glycerol-3-phosphorylcholine: 1 mug/mL; glycocholic acid- (glycine-1-13C) -hydrate: 50 mug/mL); the methanol and the acetonitrile can be packaged independently or can be mixed solution with volume ratio of 1:1.
The method of using the kit of this example is as described in example 1.
The kit can diagnose the risk of depression of the people to be detected by detecting the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in the peripheral blood plasma of the people to be detected: the risk of depression is high if the level of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is high (relative to healthy people), and low if the level of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine is low (relative to healthy people). Can be used for diagnosing clinical depression, provides effective basis for patients to take relevant therapeutic measures or decisions, and has good clinical application prospect.
Claims (6)
1. Use of a reagent for detecting 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in the preparation of a depression diagnosis kit, characterized in that the reagent is a reagent for detecting the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma;
the depression diagnosis kit diagnoses depression according to the following method: the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma is obviously positively correlated with the risk of depression, and the risk of depression is judged by detecting the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma.
2. The use according to claim 1, wherein the reagent is a liquid chromatography-mass spectrometry detection reagent.
3. The use according to claim 2, wherein the liquid chromatography-mass spectrometry detection reagent comprises 1-heptadecanoyl-glycerol-3-phosphorylcholine, glycocholic acid- (glycine-1-13C) -hydrate, methanol and acetonitrile.
4. A depression diagnostic kit comprising reagents for detecting the level of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma;
the depression diagnosis kit diagnoses depression according to the following method: the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma is obviously positively correlated with the risk of depression, and the risk of depression is judged by detecting the content of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine in human peripheral blood plasma.
5. The kit of claim 4, wherein the reagent is a liquid chromatography-mass spectrometry combination reagent.
6. The kit of claim 5, wherein the liquid chromatography-mass spectrometry reagents comprise 1-heptadecanoyl-glycerol-3-phosphorylcholine, glycocholic acid- (glycine-1-13C) -hydrate, methanol, and acetonitrile.
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| CN102978204A (en) * | 2011-11-11 | 2013-03-20 | 张康 | Detection kit and detection method for CFHR1 genotypes and CFHR1 on high-density lipoprotein and oxidation phosphorylcholine |
| CN109414446A (en) * | 2016-06-29 | 2019-03-01 | 麦克哈弗特制药有限责任公司 | Phospholipid composite |
| CN112630311A (en) * | 2020-07-09 | 2021-04-09 | 深圳云合医药科技合伙企业(有限合伙) | Metabolic markers and kits for detecting affective disorders and methods of use |
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| WO2008125940A2 (en) * | 2007-04-17 | 2008-10-23 | Pfizer Products Inc. | Nanoparticles comprising non-crystalline drug |
| WO2016115097A2 (en) * | 2015-01-12 | 2016-07-21 | Children's Medical Center Corporation | Pro-inflammatory and adjuvant functions of toll-like receptor 4 antagonists |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978204A (en) * | 2011-11-11 | 2013-03-20 | 张康 | Detection kit and detection method for CFHR1 genotypes and CFHR1 on high-density lipoprotein and oxidation phosphorylcholine |
| CN109414446A (en) * | 2016-06-29 | 2019-03-01 | 麦克哈弗特制药有限责任公司 | Phospholipid composite |
| CN112630311A (en) * | 2020-07-09 | 2021-04-09 | 深圳云合医药科技合伙企业(有限合伙) | Metabolic markers and kits for detecting affective disorders and methods of use |
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| Oxidized phospholipids inhibit endothelial cell proliferation via induction of apoptosis;Miller, C. A.等;《Journal of Investigative Medicine》;第50卷(第1期);第88A页 * |
| 抑郁症生物标志物研究进展;苑杰;严辞;刘昊;王海涛;杨美荣;王静;;国际精神病学杂志(02);第103-107页 * |
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