Application of aromatic hydrocarbon receptor in preparation of reagent or kit for early screening of traumatic sepsis

Abstract

The invention belongs to the technical field of biological medicine, and particularly discloses application of an aromatic hydrocarbon receptor as a biomarker in early risk screening of traumatic sepsis. The aromatic hydrocarbon receptor can be used as a new biomarker for predicting the occurrence of the traumatic sepsis, and the expression level of the aromatic hydrocarbon receptor is positively correlated with the occurrence of the post-traumatic sepsis, so that the aromatic hydrocarbon receptor can be used as a screening index for the occurrence risk of the traumatic sepsis. The invention provides a new technical means for early screening of traumatic sepsis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and specifically discloses the application of aryl hydrocarbon receptors as biomarkers in early risk screening of traumatic sepsis. Background Technology

[0002] Trauma is a global public health problem, causing approximately 5 million deaths annually, accounting for 10% of the global disease burden. While advancements in pre-hospital emergency care and improved intensive care systems have significantly reduced the mortality rate directly caused by early-stage trauma, complications such as sepsis and multiple organ dysfunction syndrome (MODS) remain a serious threat to patients' lives. In its latest definition, sepsis is defined as life-threatening organ dysfunction caused by a dysregulation of the host's immune response to infection. It is a common and serious complication and leading cause of death in patients with burns / trauma, infections, shock, and other acute and critical conditions. Further progression can lead to septic shock and MODS, which have high mortality rates and high treatment costs, posing a serious threat to human health. Despite significant progress in anti-infective therapy and organ support technologies in recent years, there is still no effective treatment for sepsis, and its mortality rate remains as high as 30%-70%. Therefore, addressing the development of sepsis has gradually become one of the major issues worldwide.

[0003] The progression from inflammatory response to sepsis following trauma involves numerous changes in cell function and cytokine expression, exhibiting a "cascade-like" characteristic. This process is not only difficult to predict but also progresses rapidly. Coupled with the lack of specific treatments, the diagnosis and treatment of traumatic sepsis are particularly challenging, resulting in a poor prognosis. Numerous studies have confirmed that early, precise treatment can significantly reduce mortality and improve prognosis in patients with traumatic sepsis. Therefore, early prediction of traumatic sepsis is crucial. Current research on early prediction methods for traumatic sepsis can be broadly categorized into several aspects, including trauma severity scoring systems, critical illness scoring systems, clinical signs, blood biochemical indicators, pathogenic indicators, inflammatory mediators, and metabolic changes. Clinically used biomarkers mainly include C-reactive protein and procalcitonin, but these indicators only reflect the degree of systemic inflammation at the time of testing, and the results vary greatly depending on the testing time window, resulting in low specificity. Inflammatory cytokines such as IL-6 and TNFα have short retention times in the blood, making it difficult to accurately determine the appropriate testing time. Furthermore, scoring systems based on anatomical and pathophysiological conditions, such as AIS, ISS, SOFA, and APACHE II, cannot reflect the disease progression and prognosis of trauma patients. Therefore, these indicators can only serve as auxiliary indicators for the diagnosis and monitoring of trauma sepsis. To date, no specific biomarkers for the early prediction of trauma sepsis have been discovered. Therefore, discovering new biomarkers with both high sensitivity and specificity for the early prediction of trauma sepsis is crucial for its early detection, diagnosis, and treatment. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the present invention aims to provide an application of aromatic hydrocarbon receptors as biomarkers in the early screening of trauma sepsis, thereby resolving the problems in existing technologies.

[0005] This invention first provides the use of aromatic hydrocarbon receptors in the preparation of reagents or kits for early screening of trauma sepsis.

[0006] In one embodiment of the invention, the reagent or kit detects the content of aromatic hydrocarbon receptors in a sample by enzyme-linked immunosorbent assay (ELISA).

[0007] In one embodiment of the invention, the sample is plasma.

[0008] In one embodiment of the present invention, the ELISA is a double-antibody sandwich method.

[0009] This invention also provides the use of aromatic hydrocarbon receptors as biomarkers for early screening of trauma sepsis.

[0010] In one embodiment of the invention, it includes detecting the content of aromatic hydrocarbon receptors in plasma.

[0011] In one embodiment of the invention, it is used for early screening of traumatic sepsis.

[0012] Use of reagents or kits for detecting aromatic hydrocarbon receptor content in the preparation of compositions for early screening of trauma sepsis.

[0013] Furthermore, the reagent or kit contains an antibody targeting the aromatic hydrocarbon receptor; preferably, the reagent or kit contains an antibody and a composition thereof for detecting the content of aromatic hydrocarbon receptors in a sample by enzyme-linked immunosorbent assay (ELISA).

[0014] The beneficial effects of the above-described technical solution of the present invention are as follows: The beneficial effects of this invention are as follows: This invention discovers that aryl hydrocarbon receptors can be used as a screening indicator for trauma sepsis. The expression level of aryl hydrocarbon receptors in the plasma of trauma patients with sepsis is significantly higher than that in the group without sepsis. Furthermore, by detecting the expression level of aryl hydrocarbon receptor protein in plasma, the risk of sepsis can be screened for in the early stage. The area under the curve for aryl hydrocarbon receptors is 0.804, indicating that aryl hydrocarbon receptors have good value for early screening of sepsis. Attached Figure Description

[0015] Figure 1 a is a graph showing the detection results of aromatic hydrocarbon receptor content in the plasma of healthy volunteers and trauma patients in Example 1 of the present invention; Figure 1 Figure b shows the results of aryl hydrocarbon receptor content detection in the plasma of patients with and without sepsis.

[0016] Figure 2 This diagram illustrates the early screening effect of plasma aryl hydrocarbon receptors in the risk of traumatic sepsis, based on receiver operating characteristic (ROC) analysis in Example 2 of this invention. Detailed Implementation

[0017] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0018] Example 1: Detection of plasma aryl hydrocarbon receptor levels in trauma patients and correlation analysis with the occurrence of sepsis.

[0019] Blood was collected from 243 patients with severe multiple injuries treated at the PLA Army Medical Center for Special Purposes of Medical Research and Treatment from August 2013 to June 2018, and from 13 healthy volunteers during the same period. Within 2 hours of collection, blood was centrifuged at 500g for 10 minutes at 4°C, and the plasma was separated and stored at -80°C for later use. All participants were approved by the Ethics Committee of the PLA Army Medical Center for Special Purposes of Medical Research and Treatment. Plasma aryl hydrocarbon receptor (ARH) levels were detected using an ELISA kit (FineTest, catalog number EH1522, sensitivity 0.188 ng / ml, detection range 0.313-20 ng / ml). This kit employed a double-antibody sandwich ELISA method. Anti-human ARH receptor antibodies were coated onto the ELISA plate. During the experiment, human ARH receptors in the sample or standard bound to the coated antibody, and the free components were washed away. Biotinylated anti-human ARH receptor antibody and horseradish peroxidase-labeled avidin were added sequentially. Anti-human aryl hydrocarbon receptor (ARH) antibodies bind to ARH receptors bound to the coated antibody, and biotin specifically binds to avidin to form immune complexes, while free components are washed away. A chromogenic substrate (TMB) is added; TMB turns blue under the catalysis of horseradish peroxidase, turning yellow upon the addition of a stop solution. The OD value is measured at 450 nm using a microplate reader. The concentration of ARH receptors is directly proportional to the OD450 value; the concentration of ARH receptors in the sample is calculated by plotting a standard curve.

[0020] Based on the latest diagnostic criteria for sepsis (Sepsis 3.0) published in 2016, trauma patients were divided into a sepsis group (67 cases) and a non-sepsis group (176 cases). Analysis of differences between the groups revealed ( Figure 1 a) There was no significant difference in plasma aryl hydrocarbon receptor levels between trauma patients and healthy volunteers (p>0.05), while plasma aryl hydrocarbon receptor levels in trauma patients who developed sepsis were significantly higher than those in the non-sepsis group (p<0.001). Figure 1 b). Association analysis revealed a strong correlation between plasma AHR levels and the occurrence of trauma sepsis (OR=1.21, 95%CI=1.13-1.31, p<0.001). These results indicate that peripheral plasma aryl hydrocarbon receptor levels in trauma patients are associated with the risk of trauma sepsis.

[0021] Example 2: ROC curve analysis of the early predictive value of plasma aryl hydrocarbon receptors for the risk of traumatic sepsis.

[0022] ROC curves were plotted based on the aromatic hydrocarbon receptor content values ​​detected in the plasma of trauma patients in Example 1.

[0023] Figure 2It shows a schematic diagram of the early screening effect of plasma aryl hydrocarbon receptor in the risk of sepsis by receiver operating characteristic (ROC) analysis. The results show that the area under the curve (AUC) of the aryl hydrocarbon receptor is 0.806. When the cutoff value is 1.79 ng / ml, the sensitivity is 84.2% and the specificity is 63.4%, indicating that the aryl hydrocarbon receptor has good value for early screening of sepsis.

[0024] AUC reflects the predictive efficacy (AUC = 0.5, no predictive efficacy; 0.5 < AUC < 0.7, small predictive value; 0.7 < AUC < 0.9, fairly accurate predictive value; 0.9 < AUC < 1, very accurate predictive value).

[0025] In summary, the present invention finds that the expression level of aryl hydrocarbon receptor in patients with sepsis after trauma is significantly higher than that in patients without sepsis. Through ROC curve analysis, it is found that the aryl hydrocarbon receptor can be used as a new biomarker for predicting the occurrence of sepsis.

[0026] The above is the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle described in the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Use of an aryl hydrocarbon receptor in the manufacture of a reagent or kit for early screening of trauma sepsis.

2. Use according to claim 1, characterized in that, The reagent or kit detects the content of aryl hydrocarbon receptor in a sample by enzyme-linked immunoassay (ELISA).

3. Use according to claim 2, characterized in that, The sample is plasma.

4. The use according to claim 2, characterized in that, The ELISA is a double-antibody sandwich method.

5. Use of an aryl hydrocarbon receptor as a biomarker for early screening of trauma sepsis.

6. Use according to claim 5, characterized in that, It comprises detecting the content of aryl hydrocarbon receptor in plasma.

7. Use of a reagent or kit for detecting the content of aryl hydrocarbon receptor in the manufacture of a composition for early screening of trauma sepsis.

8. The use of claim 7, wherein, The reagent or kit comprises an antibody targeting aryl hydrocarbon receptor.

9. Use according to claim 8, characterized in that, The reagent or kit comprises an antibody for detecting the content of aryl hydrocarbon receptor in a sample by enzyme-linked immunoassay (ELISA) and compositions thereof.

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