Screening and identification of potential biomarkers and establishment of the diagnostic serum proteomic model for the Traditional Chinese Medicine Syndromes of tuberculosis

Abstract

Ethnopharmacological relevance: Chemotherapy is the mainstay of modern tuberculosis (TB) control. Traditional Chinese Medicine (TCM) can enhance the effect of anti-TB drug, promote the absorption of the foci in the lung and reduce drug toxicity. In TCM, the determination of treatment is based on ZHENG (also called TCM syndrome). To establish a diagnostic model by using proteomics technology in order to identify potential biomarkers for TCM syndromes of TB.

Materials and methods: The surface-enhanced laser desorption ionization time of flight mass spectrometer (SELDI-TOF MS) combined with weak cation exchange (WCX) magnetic beads was used to screen serum samples from 71 cases of deficiency of lung yin syndrome (DLYS), 64 cases of fire (yang) excess yin deficiency syndrome (FEYDS) and 45 cases of deficiency of both qi and yin syndrome (DQYS). A classification model was established by Biomarker Pattern Software (BPS). Candidate protein biomarkers were purified by reverse phase-high performance liquid chromatograph (RP-HPLC), identified by MALDI-TOF MS, LC-MS/MS and validated by ProteinChip Immunoassays.

Results: A total of 74 discriminating m/z peaks (P<0.001) among three TCM syndromes of TB were detected. A diagnostic model for the TCM syndrome of TB based on the five biomarkers (3961.7, 4679.7, 5646.4, 8891.2 and 9416.7 m/z) was established which could discriminate DLYS, FEYDS and DQYS patients with an accuracy of 74.0%, 72.5%, and 96.7%, respectively. The candidate biomarker with m/z of 9416.7 was identified as a fragment of apolipoprotein C-III (apoC-III) by MALDI-TOF-MS and LC-MS/MS.

Conclusion: The TCM syndrome diagnostic model of TB could successfully distinguish the three TCM syndromes of TB patients. This provided a biological basis for the determination of treatment based on different TCM syndromes of TB. ApoC-III was identified as a potential biomarker for TCM syndromes of TB and revealed the biochemical basis and pathogenesis of TCM syndromes in TB.

Keywords: Apolipoprotein C-III; Proteomics; TCM syndrome; Traditional Chinese Medicine; Tuberculosis.