Title:
[Analysis of differential gene expression profile in peripheral blood of patients with chronic hepatitis B and syndromes of dual deficiency of liver and kidney yin and accumulation of dampness heat].

Abstract:
To investigate the differential gene expression profile in two typical traditional Chinese medicine (TCM) syndromes of patients with chronic hepatitis B (CHB), and to find the relationship between TCM syndromes and gene expressions.
Patients with CHB were collected from Department of Liver Diseases of Longhua Hospital in Shanghai and diagnosed as dual deficiency of liver and kidney yin syndrome (n=3) or accumulation of dampness heat syndrome (n=3). Three healthy volunteers were used as a control. Blood samples were collected before treatment. After total RNA was isolated from leukocytes, the gene expression profiles were detected by microarray. For confirming the gene expressions, a further 10 patients with dual deficiency of liver and kidney yin syndrome or accumulation of dampness heat syndrome and 10 healthy volunteers were enrolled. Their peripheral blood was used to test the gene expression by real-time reverse transcription-polymerase chain reaction (RT-PCR) which was differently expressed by microarray.
Microarray analysis identified that there were significant differences in gene expression between patients with dual deficiency of liver and kidney yin syndrome and patients with accumulation of dampness heat syndrome, as well as between CHB patients and healthy volunteers. Patients with dual deficiency of liver and kidney yin syndrome and accumulation of dampness heat syndrome were fully distinguished by clustering of 403 differentially expressed genes (P<0.05). A total of 239 genes were significantly differentially expressed (absolute value of foldchange≥2, P<0.05), among which 142 were up-regulated and 97 were down-regulated. Genes specially regulated by the dual deficiency of liver and kidney yin syndrome were mainly related to peroxide activity and stem cell maintenance; genes specially regulated by the accumulation of dampness heat syndrome were mainly related to cytokines, immune and inflammatory responses. ATP-binding cassette, sub-family C (CFTR/MRP), member 3, peroxisome proliferator-activated receptor α and phosphatidic acid phosphatase type 2A mRNA expressions detected by RT-PCR were similar to the results of microarray.
Differential gene expressions are observed in CHB patients with dual deficiency of liver and kidney yin syndrome and accumulation of dampness heat syndrome, indicating that there is molecular foundation in the classification of TCM syndrome. The preliminary findings of this study suggest that TCM syndrome types are related to gene expression.