Lymphocyte Apoptosis In Vitro Methotrexate And Alkylating Agents In Combination In Patients With Rheumatoid Arthritis

Objectives: To observe the apoptosis effect of combination therapy with chlormethine and methotrexate on lymphocyte cells taken from rheumatoid arthritis patiences in vitro. Methods: We observed morphological changes by immuno- fluorescence microscope. Apoptosis of lymphocyte was analyzed by FCM (Annexin-FI TC/PI stained). Results: 1. Apoptosis ratio of lymphocyte show a little increasing trend with the increased concentration of methotrexate within limited range (6-60 μmol/l). 2. Apoptosis ratio of lymphocyte show a linearly increasing trend with the increased concentration of chlormethine. 3. When phytahematoagglutinin-actived lymphocytes were incubated with a certain concentration drugs for 72 hours, apoptosis ratio of lymphocyte in Methotrexate group (concentration 0.6μmol/l) and in chlormethine group (concentration10μmol/l) are no significant differences from the control group (P>0.05). The Apoptosis ratio of combination group is more significant increased than the other groups (P<0.05). While necrosis ratio of the four groups are no significant differences among them (P>0.05). Conclusion:Combination therapy with chlormethine and Methotrexate has cooperation effect on lymphocyte in apoptosis in vitro.