Immunoregulation of Chloroform Component Extracted from Erteng Mixture on T Lymphocytes in Rheumatoid Arthritis / 广州中医药大学学报

ABSTRACT

[Objective] To investigate the effect of Erteng Mixture (EM) on activated T-cell CD69 expression and Jurkat cell cycle in rheumatoid arthritis (RA) and to explore its possible cellular immunoregulation mechanism. [Methods] Jurkat cells were cultured with different kinds of components extracted from EM. After 72 hours, proliferation rate of Jurkat cell were detected with thiazolyl blue colorimetry (MTT) to screen the active component. Whole blood from RA patients were incubated in RPMI-1640 culture with various concentrations of chlorofonn component extracted from EM or with Genistein. Thirty minutes later, phytahematoagglutinin (PHA) were added successively into the culture. After 24 hours, CD69 expression rate of T lymphocytes activated by PHA was analyzed by flow cytometry. Jurkat cells were cultured with various concentrations of chloroform component extracted from EM and with methotrexate ( MIX) for 48 hours. After then, Jurkat cell cycle was analyzed by flow cytometry. [ Results ] Chloroform component extracted from EM had the strongest inhibitive effect on proliferation of Jurkat cells. Chloroform component (10 mg/L and 20 mg/L) could markedly inhibit CD69 expression on PHA-activated T lymphocytes. Chloroform component 10 mg/L blocked Jurkat cell cycle at G1 phase, which differed from MTX blocking Jurkat cell cycle at S phase. [Conclusion] Chloroform component extracted from EM could significantly inhibit the early activation of CD4 T lymphocytes in RA and inhibit the energy and material synthesis in T-cell, thus hindering DNA synthesis and decreasing the cellular G2 phase transformation. This study will provide an experimental basis for clinical application of EM in treating RA.