Screening And Identification Of Potenial Biomarkers For Multidrug-resistant Tuberculosis And Its Traditional Chinese Medicine Syndromes

At present,tuberculosis is still the most deady infectious disease in the world,especially multidrug-resistant tuberculosis?MDR-TB?is a key problem in global tuberculosis control Lack of effective early diagnosis methods,as well as long treatment cycles,large side effects,and patient intolerance,make the MDR-TB epidemic unable to be effectively controlled Traditional Chinese Medicine?TCM?treatment of MDR-TB has a good attenuating and synergistic effect.However,the lack of objective biological evidence for the classification and diagnosis of MDR-TB TCM syndromes has resulted in low efficacy of TCM treatment for MDR-TB.In first part study,we investigated long non-coding RNAs?IncRNAs?in plasma of MDR-TB patients,DS-TB?drug-sensitive tuberculosis?patients,and HC?healthy control?individuals and the potential diagnostic value for MDR-TB,using microarray,bioinformatics analysis and real-time quantitative polymerase chain reaction?RT-qPCR?methods.In second part study,isobaric tags for relative and absolute quantification-coupled twodimensional liquids chromatography-tandem mass spectrometry?iTRAQ-2DLC-MS/MS?combined with bioinformatics analysis method was applied to identify and screen the differentially expressed serum proteins?DEPs?,followed by the biological functional analysis.Additionally,DEPs among three different TCM syndromes of MDR-TB were verified and analyzed by enzyme-linked immune sorbent assay?ELISA?.Finally,a receiver operating characteristic?ROC?curve was performed to estimate the diagnostic ability of DEPs.The first part study found,compared with the DS-TB group,that a total of 543 IncRNAs?195 mRNAs?differentially expressed in the MDR-TB group,of which 181?63 mRNAs?were up-regulated and 362 were down-regulated?132 mRNAs?;Compared with the HC group,a total of 699 IncRNAs?231 mRNAs?were found to be differentially expressed in the MDR-TB group,including 345?123 mRNAs?up-regulated IncRNAs and 354 down-regulated IncRNAs?108 mRNAs?.Gene ontology?GO?,Kyoto Encyclopedia of Genes and Genomes?KEGG?and coding-noncoding co-expression?CNC?analyses showed that functions of differentially expressed IncRNAs were mainly enriched in the metabolism and regulation of substances,such as the fatty acid degradation pathway and the lysine degradation pathway.RT-qPCR confirmed that three IncRNAs were up-regulated in the MDR-TB group,including NR₁10750?P=0.036?,uc.212-?P=0.011?,and NR₁31237?P=0.008?.The second part study found,A total of 71 proteins were differentially expressed in the three different TCM syndrome groups of MDR-TB:pulmonary Yin deficiency?PYD?syndromegroup,Hyperactivity of Fire due to Yin deficiency?HFYD?syndrome group and deficiency of Qi and Yin?DQY?syndrome group.The expression level of transforming growth factor-beta-induced protein ig-h3?TGFBI?waslower in PYD syndrome group?P=0.002?.Pro-protein convertase subtilisin/kexin type 9?PCSK9?overexpressed in HFYD syndrome group?P<0.0001?.C-C motif chemokine 14?CCL14?expression level was reduced inDQY syndromegroup?P=0.004?.Therefore,this study demonstrated that serum TGFBI,PCSK9 and CCL14 are new potential biomarkers of PYD syndrome,HFYD syndrome and DQY syndrome for MDR-TB respectively.The first part study reveals the potential value of IncRNAs as biomarkers for early diagnosis of MDR-TB and the underlying mechanisms of these aberrantly expressed IncRNAs in the pathogenesis of MDR-TB.The first part study,this study provids biological basis for MDR-TB TCM syndrome classification and of great significance to take different treatment options for MDR-TB different TCM syndrome candidates.