Effects Of Alkaloid Sinomenine On Expression Of CD69 And Apoptosis Of CD4～+ T Lymphocytes Model Of Human Peripheral Blood In Vitro

Objectives: To evaluate the immunosuppressive efficacy and outcome of alkaloid sinomenine(SIN) on the CD4+ T lymphocytes model of human peripheral blood.The chosen indicators are the expression of CD69 and apoptosis of CD4+T cells proliferation model of human peripheral blood.The aim of this study is to provide theoretical foundation for clinical to apply the SIN for immunosuppressive therapy.Methods:Establishment of the CD4+T lymphocytes model of human peripheral blood: 1) Withdrawaled 50ml vein blood of one healthy adult male volunteer,adding 500U heprin to protect the blood from coagulation;2)Isolated mononuclear cells from the vein blood cells by lymphocytes separation medium;3) Isolated CD4+T lymphocytes from the mononuclear cells suspension by immunological magnetic beards;4) The isolated CD4+ T lymphocytes were cultured with RPMI-1640/10%FCS complete culture medium;3)The CD4+T lymphocytes were proliferated with Phytohemagglutinin(PHA,lastconcentration:10ug/ml)and IL-2 (lastconcentration:100 U/ml) for 7 days .The proliferated CD4+ T lymphocytes suspension were divided averagely into 5 groups at random whose cell concentration was 5×105 and volume was 10 ml: 1)Group contRast:No medicine was added to cell culture medium; 2) Group genisein:Genistein solution whose last concentration was 50 umol/L was added to cell culture medium; 3) Group low-concentration SIN: SIN solution whose last concentration was 10 umol/L was added to cell culture media;4) Group middle-concentratione SIN: SIN soluton whose last concentration was 200 umol/L was added to cell culture media;5) Group high-concentrationed SIN : SIN soluton whose last concentration was 1000 umol/L was added to cell culture media. 6 hours later, anti-CD3 mAb solution whose final concentration was 1ug/ml was added to cell culture media of...