阅读说明：本技术 长链脂肪酸辅酶a连接酶1在诊断或预测子痫前期中的应用 (Application of long-chain fatty acid coenzyme A ligase 1 in diagnosis or prediction of preeclampsia ) 是由 蔡伟 徐忠伟 牛秀珑 陈少伯 李玉明 于 2019-11-26 设计创作，主要内容包括：本发明公开了长链脂肪酸辅酶A连接酶1的标记试剂在制备用于诊断孕妇是否患有子痫前期或者用于预测孕妇是否具有患子痫前期的风险的试剂盒中的应用,属于生物医学领域。本发明还公开了用于诊断孕妇是否患有子痫前期或者用于预测孕妇是否具有患子痫前期的风险的试剂盒,包括长链脂肪酸辅酶A连接酶1的标记试剂。本发明为妊娠期高血压疾病子痫前期的诊断和预警提供了一个新的生物标记物,具有重要的临床价值。(The invention discloses an application of a labeled reagent of long-chain fatty acid coenzyme A ligase 1 in preparation of a kit for diagnosing whether a pregnant woman has preeclampsia or predicting whether the pregnant woman has the risk of preeclampsia, and belongs to the field of biomedicine. The invention also discloses a kit for diagnosing whether the pregnant woman has preeclampsia or predicting whether the pregnant woman has the risk of preeclampsia, which comprises a labeling reagent of the long-chain fatty acid coenzyme A ligase 1. The invention provides a new biomarker for diagnosis and early warning of preeclampsia of hypertensive disorders in pregnancy and has important clinical value.)

1. Application of a labeled reagent of long-chain fatty acid coenzyme A ligase 1 in preparation of a kit for diagnosing whether a pregnant woman has preeclampsia or predicting whether the pregnant woman has risk of preeclampsia.

2. The use according to claim 1, wherein the labeling agent is an antibody.

3. A kit for diagnosing whether a pregnant woman has preeclampsia or for predicting whether a pregnant woman is at risk for preeclampsia, comprising a labeled reagent for long chain fatty acid coa ligase 1.

4. The kit of claim 3, wherein the labeling reagent is an antibody.

5. The kit of claim 3 or 4, wherein the kit is used to diagnose or predict a long chain fatty acid coenzyme A ligase 1 expression level in a sample from the pregnant woman who has or is at risk of developing preeclampsia if the long chain fatty acid coenzyme A ligase 1 expression level in the sample exceeds a threshold value.

6. The kit of claim 5, wherein the maternal sample is a serum sample and/or a placental sample.

7. The kit of claim 5, wherein the threshold is an upper limit of a 95% confidence interval for long chain fatty acid coenzyme A ligase 1 expression in a medically statistically significant number of normal maternal samples.

8. The kit of claim 5, wherein the threshold is a lower limit of 95% confidence interval for long chain fatty acid coenzyme A ligase 1 expression in a medically statistically significant number of samples of pregnant women with preeclampsia.

9. The kit of claim 5, further comprising a positive control and a negative control.

10. The kit of claim 9, wherein the positive control is a sample from a pregnant woman diagnosed with preeclampsia.

Technical Field

The invention belongs to the field of biomedicine, and particularly relates to application of long-chain fatty acid coenzyme A ligase 1 in diagnosis or prediction of preeclampsia.

Background

Preeclampsia (PE), which is an important subtype of hypertensive disorders in pregnancy, accounts for more than half of hypertensive disorders in pregnancy, and is one of the main causes of death of pregnant and lying-in women worldwide. The etiology and pathogenesis of PE have not been fully elucidated, and current research shows that PE is a placenta-derived disease with multiple factors and multiple mechanisms acting together, and has similar pathophysiological mechanisms with cardiovascular diseases (CVD), such as oxidative stress, vascular endothelial injury, insulin resistance, lipid metabolism disorder and the like. PE not only causes adverse events of pregnancy to threaten the life of the mother and the child, but also is a risk factor of CVD diseases of the mother and the child in the two generations of the future, and a plurality of researches and meta-analysis show that the risk of the PE female in the future suffering from the CVD diseases such as myocardial infarction, cerebral apoplexy and the like is up to several times higher, the CVD morbidity is positively correlated with the severity of the PE, and the risk of hypertension, CVD and metabolic diseases of the child is also obviously increased.

The PE diagnosis standard is that the systolic pressure is more than or equal to 140 mmHg and/or the diastolic pressure is more than or equal to 90 mmHg after 20 weeks of gestation, and is accompanied by any one of the following: urine protein is more than or equal to 0.3g/24h, or the ratio of urine protein to creatinine is more than or equal to 0.3, or random urine protein is more than or equal to (+) (the detection method when the urine protein can not be quantified); proteinuria-free but with any organ or system involvement: abnormal changes in the vital organs such as heart, lung, liver, kidney, etc., or in the blood system, digestive system, and nervous system, and involvement of placenta-fetus, etc. From the above diagnostic criteria it can be seen that a definitive diagnosis of PE requires 20 weeks after gestation and that the measures for treatment after diagnosis are effective. Hypertensive disorder during pregnancy: international guidelines for the study of hypertension in gestational period (2018) and chinese guidelines for diagnosis and treatment of hypertensive disorders in gestational period (2015) indicate that the main treatment of PE is to prevent convulsion, lower blood pressure with indications, induce diuresis, calm, closely monitor maternal and fetal conditions, prevent and treat serious complications, and terminate pregnancy at appropriate time.

At present, under the conditions that PE patients bring about mortality of pregnant and lying-in women all over the world and perinatal adverse events are high, the only effective treatment way for PE is to terminate pregnancy and deliver placenta, and premature termination of pregnancy can bring about adverse fate of premature infants and lying-in women, thus increasing the burden of families and society. Thus, if PE can be predicted in advance, and intervention is given early, it is possible to reduce the incidence of PE, reduce pregnancy failure outcomes, and reduce pregnancy CVD risk exposure. To this end, there is currently a great deal of PE prediction research and validation work, mainly to find new serum biomarkers and build prediction models for early prediction, however, until now, methods for reliably predicting all preeclampsia in the early and middle trimester of pregnancy are lacking. Even the few potential markers that appear (e.g., sFlt-1/PlGF ratio) are only valuable in extending the expected treatment late in pregnancy and cannot be used to predict PE development; some markers have insignificant or unstable predictive efficacy and cannot be clinically used. There are studies using maternal risk factors, blood pressure, PlGF and uterine artery Doppler combinations that might predict pre-term type preeclampsia (< 37 weeks), but with higher false positive or false negative results in terms of efficacy. Therefore, the search for sensitive preeclampsia biomarkers is very important for early warning and diagnosing preeclampsia caused by hypertensive disorders in pregnancy.

Disclosure of Invention

In order to solve the above technical problems, the present invention aims to provide a biomarker for preeclampsia. For this reason, the inventors have conducted extensive research and unexpectedly found that Long-chain fatty acid coenzyme a ligase 1 (Long-chain-fat-acid-CoA ligase 1, ACSL 1) is significantly associated with preeclampsia, and further confirmed, thereby completing the present invention.

The invention provides application of a labeled reagent of long-chain fatty acid coenzyme A ligase 1 in preparation of a kit for diagnosing whether a pregnant woman has preeclampsia or predicting whether the pregnant woman has the risk of preeclampsia.

ACSL1 is an isozyme of the family of long chain fatty acid coenzyme A ligases. Although substrate specificity, subcellular localization, and tissue distribution vary, all isozymes of this family convert free long-chain fatty acids to fatty acid acyl-coa esters, thereby playing a key role in lipid biosynthesis and fatty acid degradation.

Further, the labeling reagent is an antibody. In a specific embodiment of the invention, the antibody is a rabbit anti-ACSL 1 antibody.

In a second aspect, the invention provides a kit for diagnosing whether a pregnant woman has preeclampsia or predicting whether a pregnant woman is at risk of preeclampsia, comprising a labeled reagent for long chain fatty acid coenzyme A ligase 1.

Further, the labeling reagent is an antibody. In a specific embodiment of the invention, the antibody is a rabbit anti-ACSL 1 antibody.

In some embodiments of the invention, the kit performs diagnosis or prognosis by detecting whether the expression level of long chain fatty acid coa ligase 1 in a sample from the pregnant woman exceeds a threshold, and if the expression level of long chain fatty acid coa ligase 1 in the sample exceeds the threshold, the pregnant woman suffers from or is at risk of pre-eclampsia.

In some embodiments of the invention, the threshold is the upper limit of the 95% confidence interval for the expression of long chain fatty acid coenzyme a ligase 1 in a medically statistically significant number of normal maternal samples.

In other embodiments of the invention, the threshold is the lower limit of the 95% confidence interval for the expression of long chain fatty acid coenzyme A ligase 1 in a medically statistically significant number of samples of pregnant women with preeclampsia.

Further, the kit further comprises a positive control substance and a negative control substance.

In some embodiments of the invention, the positive control is a sample from a pregnant woman diagnosed with preeclampsia.

In the present invention, the maternal sample is a serum sample and/or a placental sample.

The early warning and diagnosis marker for the preeclampsia of the hypertensive disorder in the gestation period has the characteristics of good specificity, high sensitivity, good reproducibility, good patient compliance and the like. Experiments prove that the ACSL1 can be used as an early warning and diagnosis marker for the pregnancy hypertension disease preeclampsia.

The invention has the advantages of

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a new biomarker for diagnosis and early warning of preeclampsia of hypertensive disorders in pregnancy and has important clinical value.

The result of mass spectrometry is used for confirming that the ACSL1 expression level in the serum sample of the patient with the pregnancy hypertensive disease preeclampsia is obviously higher than that of the patient with the normal pregnancy group.

The relative level of ACLS1 in the placental tissues of patients with hypertensive disorders of pregnancy preeclampsia was significantly higher than the sexual-pair level of ACLS1 in the placental tissues of normal pregnancy (p < 0.0001), with a sensitivity of 91.13% and a specificity of 93.67%.

The relative level of ACLS1 in the serum of patients with hypertensive disorders of pregnancy pre-eclampsia was very significantly higher than the sex-pair level of ACLS1 in the serum of patients with normal pregnancy (p < 0.0001).

The ACSL1 can be used for early warning screening of patients with pregnancy-induced hypertension preeclampsia, and has the advantages of accurate result, simple operation and great popularization and application value.

Drawings

FIG. 1 shows the mass spectrum detection results in example 1.

Figure 2 shows the proteomics signal path detection results in example 1.

FIG. 3 shows the results of immunohistochemistry in example 2.

Fig. 4 shows the optical density value analysis result in example 2.

FIG. 5 shows the ACSL1 in example 3 as the receiver operating characteristic curve (ROC curve), 1 represents normal human, 2 pre-eclamptic maternal patient.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects solved by the present invention more apparent, the present invention is further described in detail below with reference to the following embodiments.