Expression of PYCARD gene transcript variant mRNA in peripheral blood mononuclear cells of primary gout patients with different Chinese medicine syndromes.

ABSTRACT

OBJECTIVE: To study the expression level and role of apoptosis-associated speck-like protein containing a caspase recruitment domain (PYCARD) gene transcript variant mRNA in peripheral blood mononuclear cells (PBMCs) of primary gout (PG) patients with different Chinese medicine (CM) syndromes. METHODS: The expressions of PYCARD gene transcript variant mRNA and interleukin-1ß (IL-1ß) mRNA in PBMCs were investigated in 96 PG patients with acute phase (APPG, 44 cases) and non-acute phase (NAPPG, 52 cases) and 30 healthy controls (HCs) by reverse transcription-polymerase chain reaction (PCR) and/or realtime quantitative PCR. PYCARD and nuclear factor-κB (p50) [NF-κB (p50)] protein was detected by Western blot in PBMCs respectively. IL-1ß, IL-4 and IL-10 protein levels in plasma of HCs and PG patients were measured by enzyme-linked immuno sorbent assay. RESULTS: The main CM syndromes in APPG patients were obstruction of dampness and heat syndrome (ODHS, 36.36%) and intermingled phlegm-blood stasis syndrome (IPBSS, 27.27%), while in NAPPG patients were Pi (Spleen)-deficiency induced dampness syndrome (PDIDS, 40.38%) and qi-blood deficiency syndrome (QBDS, 26.92%). It showed statistical significances of the expressions of PYCARD gene and its transcript variant mRNA, the protein of PYCARD and NF-κB (p50) and the plasma IL-1ß, IL-4 and IL-10 in APPG, NAPPG, ODHS, IPBSS, PDIDS and QBDS groups, compared with the HC group respectively (P<0.05 or P<0.01). There were also significant differences of mRNA expressions of PYCARD-1 and PYCARD-2 as well as protein expressions of IL-1ß, IL-4 and IL-10 among the 4 CM syndromes groups (P<0.05 or P<0.01). Correlation analysis showed positive correlation between the mRNA expressions of PYCARD-1 gene transcript variant and IL-1ß in APPG patients (r=0.3088, P=0.0183). CONCLUSION: PYCARD gene and its transcript variant may play a critical and regulative role in the inflflammatory response of PG patients with different phases and CM syndromes.