Study On Serum Protein Expression Profile Of Asthma And ABPA

Research background: Asthma is a common chronic airway inflammation with variable and reversible airflow restrictions,of which allergic bronchial bronchial psycosis(ABPA)is one of the subtypes.The clinical manifestations of the two are relatively similar,possibly recurrent wheezing,shortness of breath,chest tightness or coughing,lung shadow and bronchial dilation.However,the cause of the former is complex and often unknown,while the latter is an immune allergy caused by smoking mold.In China,the incidence of asthma is 4.2%.Although ABPA patients account for only 2% of asthma patients,there are still a large number of new cases each year,based on China’s large population.Many patients are left out or misdiagnosed due to the lack of specific diagnostic and differential diagnostic criteria.However,since clinical diagnosis and treatment rely primarily on the personal knowledge and experience of physicians,they are looking for new,specific and highly sensitive biomarkers to assist in the early diagnosis of asthma and ABPA patients,especially those with asthma.It is of great significance of basic research and clinical application value.Liquid biopsies use blood and urine samples as test subjects for low-risk in vitro diagnostics of diseases,such as dynamic diagnosis,disease process monitoring,efficacy assessment and prognostic intervention.In recent years,because of its irreplaceable advantages,it has been paid more and more attention and promotion in clinical applications.In this study,the advantages of this technique were used to detect low-abundance proteins in three groups of normal control,asthma and ABPA serum samples,and to screen for a group of proteins and diseases that may be expressed differently in terms of incidence.Analysis to find new protein molecular markers with diagnostic and prognostic value for asthma and ABPA lesions.Methods: The study first collected three serum samples from normal control subjects,asthma and ABPA patients,and used the i TRAQ method of removing high-abundance structure and nonspecific proteins,respectively,to obtain the proteins between the two diseases and the normal control group,and the differences between the two disease groups.Subsequently,a variety of bioinformatics software and analytical platforms were used to try to uncover the possible functioning of this batch of disease-related candidate proteins.In addition,through the chip detection technology of serum protein,the candidate markers of diagnostic significance are excavated and verified in the known protein information from another detection angle.In turn,we used the ELISA method,which can be quantitatively,to verify the expression levels of 15 candidate proteins selected from the above-mentioned different bio-letter analysis.Furthermore,the analysis of pre-proteomics was once again validated by the use of multiple reaction monitoring(MRM)technology.Results:(1)Through SDS-PAGE glue testing,it is clearly visible that the sample of high abundance protein removed from serum was compared with the same untreated sample,showing a significant weakening of the band of high abundance protein.At the same time,there are many more low-abundance protein bands.It was shown that when the sample was treated,the sample could show more functional proteins,(2)618 confidence proteins were identified by quantitative detection and quality analysis of i TRAQ method,and then the samples were compared with the use of Omicsbean’s platform software: the asthma group compared with the normal control group,found23 different proteins,the ABPA group and the normal control group,found 35 differences in proteins,and ABPA group and asthma.And in the two groups of two different proteins obtained after the comparison,23 different proteins appeared repeatedly,indicating that they had common expression in asthma and ABPA diseases,and(3)after large-flow,inflammatory reaction-related commercial protein chip testing,a total of 440 known certain characteristics and function differences proteins were found.The model was compared with Ray Biotech’s platform software: 7differential proteins were found in the asthma group with the normal control group,15 differential proteins were found in the ABPA group with the normal control group,and 15 differential proteins were found in the ABPA group with asthma.(4)From the above two proteomic test results,the first 10 differential proteins with good correlation with disease lesions were selected,and ELISA was verified.Among them,the expression trend and quantitative analysis results of CSF1 differential proteins are consistent with the screening results of protein chips,indicating that CSF1 may have the potential to be an auxiliary diagnosis of ABPA and asthma,and(5)due to the lack of specific antibodies in some of the candidate proteins to be validated,the study used MRM technology to quantitatively verify the candidate proteins.First,three groups of whole protein banks were established and compared with candidate proteins of i TRAQ results that the protein chip failed to screen,and the asthma group or ABPA group were identified with the normal control group,and the 38 differential proteins in the ABPA group and asthma group were quantitatively validated with the results of the i TRAQ experimental analysis.The MRM results showed that there were significant differences between the new 8 target proteins in asthma patients and normal controls,the significant differencebetween between 4 target proteins in ABPA patients and normal controls,the difference between 8 target proteins in ABPA patients and asthma,and(6)the RESULTs of a series of HORIZONTAL mapping of these target proteins,which resulted in: a set of SINF2,ITIH3,HPANDA protein combination,It may be an ideal asthma diagnostic marker,while the combination of SERPINF1 and SERPINF2 may assist in the diagnosis of ABPA disease,and the combination of CNDP1 and 1 protein marker(i.e.,proteins consisting of SERPINF2,ITIH3,HP and SERP INC14)may have the potential to identify ABPA and asthma.These candidate proteins show high sensitivity and specificity,and it is worth digging deep into their biological and pathophysiological significance,and converting them to clinical applications as soon as possible.Conclusion: When serum samples are first treated with high abundance protein,i TRAQ detection method and co-protein chip for proteomic sie sieve and analysis,the advantages of high throughput,strong repeatability and stable results can be obtained.When further on the differential proteins associated with disease lesions,after multi-level comprehensive analysis and prediction of bioinformatics,more credible molecular markers than a single proteomic strain technique can be screened.Then,when e LISA and MRM are used to verify the candidate protein several times,the verification complements each other in the technical means,and the experimental results are more reliable.Significantly,this study found that a group of protein combinations may have the value of asthma diagnostic markers,another group of proteins that have been optimized to have the potential for ABPA diagnostic,and a group of proteins that may be markers of differential diagnosis for both ABPA and asthma.These new findings have led medical scientists to deepen their understanding of the occurrence,development and prognosis of asthma and ABPA diseases,and to open up new directions and paths for the precise diagnosis and treatment of both diseases,as well as the development of new programs and new technologies.