Classification of Traditional Chinese Medicine Syndromes in Patients with Chronic Hepatitis B by SELDI-Based ProteinChip Analysis

Abstract

Traditional Chinese medicine (TCM) syndrome, also called ZHENG, is the basis concept of TCM theory. It plays an important role in TCM practice. There are excess and deficiency syndromes in TCM syndrome. They are the common syndromes in chronic hepatitis B (CHB) patients. Here we aim to explore serum protein profiles and potential biomarkers for classification of TCM syndromes in CHB patients. 24 healthy controls and two cohorts of CHB patients of excess syndrome (n = 25) or deficiency syndrome (n = 19) were involved in this study. Protein profiles were obtained by surface-enhanced laser desorption ionization time-flight mass spectrometry (SELDI-TOF/MS) and multiple analyses were performed. Based on SELDI ProteinChip data, healthy controls and CHB patients or excess and deficiency syndromes in CHB patients were obviously differentiated by orthogonal partial least square (OPLS) analysis. Two significant serum proteins (m/z 4187 and m/z 5032) for classifying excess and deficiency syndromes were found. Moreover, the area under the receiver operating characteristic (ROC) curve was 0.887 for classifying excess and nonexcess syndrome, and 0.700 for classifying deficiency and nondeficiency syndrome, respectively. Therefore, the present study provided the possibility of TCM syndrome classification in CHB patients using a universally acceptable scientific approach.

Figure 1

Representative protein profiles of serum samples of healthy controls and patients with CHB of excess symptom and deficiency syndrome. Protein peak spectrum of serum was analyzed by the SELDI-TOF/MS system, and representative protein peaks within m/z 0–1,5000 of three groups are shown (a). Statistically significantly different peaks between excess syndrome and deficiency syndrome are shown in the enlarged view, m/z 3168 on the left and m/z 4187 on the right (b).

Figure 2

PCA score plot and OPLS score plots of 25 CHB patients of excess syndrome (▪), 19 CHB patients of deficiency syndrome (∆), and 24 healthy controls (∗) based on the serum protein profiling detected from SELDI-TOF/MS or the clinicopathological data of each individuals. (a) PCA score plot among the control group and CHB groups of excess syndrome and deficiency syndrome; OPLS score plots (b) among the control group and CHB groups of excess syndrome and deficiency syndrome and (c) between excess syndrome group and deficiency syndrome group. (a)–(c) Models of score plots were constructed by the data from SELDI-TOF/MS. (d) Another OPLS score plot among the three groups using clinical parameters.

Figure 3

Box-plots for protein peak comparison between TCM syndrome groups. Proteins m/z 1216 (a), m/z 3168 (b), and m/z 4187 (c) were in lower abundance in excess syndrome group than those in deficiency syndrome one, while protein m/z 5032 (d) was in higher abundance.

Figure 4

Diagnostic potential of the two marker proteins (m/z 4187 and m/z 5032) using binary logistic regression method with the data from different TCM syndromes in CHB patients. 88% of excess syndrome patients and 73.7% of deficiency syndrome patients were correctly discriminated (cutoff value: 0.5).

Figure 5

ROC curve for classification of two different TCM syndromes in CHB patients. It was generated combining the peak values of m/z 4187 and m/z 5032. (a) ROC curve for classification of excess syndrome and non-excess syndrome. AUC (area under the curve) = 0.887. (b) ROC curve for classification of deficiency syndrome and nondeficiency syndrome. AUC = 0.700.