The Study On The Verification Of Differential Proteomic Markers Between Syndrome Of Spleen-Kidney Yang Deficiency And Heat-toxin Stagnation In Asymptomatic Period Of AIDS

BackgroundThe asymptomatic period of AIDS is an important period of the combination of Chinese and Western treatment intervention.It is of great significance to explore the syndrome principle and characteristics of TCM in this period to improve the level and curative effect of TCM syndrome differentiation and treatment.Due to the complex pathogenesis of AIDS and the variety of pathological products,there has not been a recognized and unified syndrome classification,which has affected the progress of the application of traditional TCM in AIDS prevention and treatment.The application of biomarkers in disease screening,diagnosis and evaluation of curative effect is of great significance.In this study group,iTRAQ technology was used to screen out the significant differences in HIV/AIDS symptoms and spleen-kidney yang deficiency patients.Among them,the hot toxic accumulation test group screened 184 proteins with significant differences(the difference was greater than 1.2),of which 94 were upgraded and 90 were downgraded.It was found to be mainly involved in the regulation of defense responses,acute inflammatory responses,and normal regulation of immune effects,regulation of leukocyte immune-mediated,response to external stimuli,etc.;spleen-kidney-yang deficiency group screened 174 significant difference proteins(difference greater than 1.2)Of these,79 were raised and 95 were adjusted downward.The functions involved mainly include oxidoreductase activity,regulation of endothelial cell proliferation,peroxidase activity,regulation of intracellular protein kinase levels,antioxidant activity,regulation of signal transduction,and catalytic activity.Based on the pre-screening results of the research group,we further verify the protein which has a high correlation with the disease syndrome,and provide the basis for revealing the connotation of TCM syndrome scientifically and objectively.ObjectiveWestern Blod technique was used to further verify the protein of high specificity protein in the pre-screening results,and the expression of the target protein was compared with the screening results and was to obtain the characteristic protein spectrum of heat toxin accumulation syndrome and spleen and kidney yang deficiency syndrome in asymptomatic period of AIDS.In order to provide scientific objective basis for the diagnosis of TCM syndromes of AIDS,the biological basis and scientific connotation of the two syndromes are excavated more deeply.MethodsThe high specific protein was selected in the pre-screening of the study group,among them,four target proteins with significant differences were expressed,and the target proteins of heat toxin accumulation syndrome were:apoD、FN1、Gpx1;the target protein of spleen and kidney yang deficiency syndrome was: ApoE.The venous blood was collected from three groups of patients,and the protein of the ApoD、GPX1、FN1 and apoE of serum samples were determined by Western blotting technique,and its gray value was analyzed by image J software then compare the gray value of the target protein with the corresponding albumin gray value in each swimming lane to obtain the relative gray value and imported into GraphPad Prism 5 software respectively.Fisher accurate probability method was used to count the data.Mean± standard deviation((?)±s)is used to express normal distribution data.P < 0.05 was used as the difference standard.Results1.The results of clinical verification in the group of heat toxin accumulation syndrome in the asymptomatic period of AIDS were as follows: the expression of FN1 was down-regulated in the syndrome of heat toxin accumulation in the asymptomatic period and the expression of Gpx1 was down-regulated in the asymptomatic period of AIDS,all of which were consistent with the pre-screening results.The expression of apoD was up-regulated in the asymptomatic period of AIDS with heat toxin accumulation syndrome(P < 0.05),which was not consistent with the pre-screening results.2.Observation of asymptomatic period of AIDS spleen and kidney Yang deficiency syndrome group the results of verification results showed that the expression of small cell ApoE was down-regulated(P < 0.05),which was consistent with the pre-screening results.Conclusions1.This study verifies FN1 and Gpx1 which have a certain correlation with the syndrome of heat toxin accumulation in the asymptomatic period of AIDS,which may be a potential biomarker of heat toxin accumulation syndrome in the asymptomatic period.It may involve stress response,humoral immune response,complement activation and innate immunity.2.This study concluded that ApoE may be a potential biomarker for the syndrome of spleen and kidney yang deficiency in asymptomatic period of AIDS,and the results of validation are consistent with the results of iTRAQ screening.It is suggested that ApoE may be the characteristic protein of spleen and kidney yang deficiency syndrome in asymptomatic period of AIDS,which is involved in human immunity and inflammation.3.In this study,we obtained the differentially expressed proteins in the asymptomatic phase of the hot toxic accumulation syndrome and asymptomatic spleen-kidney yang deficiency,and proved the objective biological basis of AIDS syndromes from the perspective of proteomics.