Effects Of Alkaloid Sinomenine On Expression Of IL-2, P-ERK And Apoptosis Of CD4～+T Lymphocytes Model Of Human Peripheral Blood In Vitro

Objectives To evaluate the immunosuppressive efficacy and outcome of alkaloidsinomenine(SESI) on the CD4~+ T lymphocytes model of human peripheral blood. The chosen indicators are the expression of apoptosis of CD4~+ T cells, P-ERK and IL-2.The aims are to study its synergistic effect in combination with immunosuppressant Cyclosporin A(CsA) and provide theoretical foundation for clinical to apply the SIN for immunosuppressive therapy.Methods Establishment of human CD4~+ T lymphocyte models of peripheralblood:(1 ), 50ml elbow vein blood in three healthy adulted male volunteer was drawed, adding 500U heprin to protect the blood from coagulation;(2), PBMCs were isolated from the vein blood with lymphocytes separation medium;(3), CD4~+ T lymphocytes wereisolated from PBMC suspension with immunomagnetic beads and cultured(37℃,5%CO2) with 10%FCS RPMI-1640;(4), The CD4~+T lym- phocytes were proliferated by PHA(final concentration: 5ug/ml) and IL-2 (final concentration: 100 U/ml) for 7 days . After wash it three times by BPS,to treat as followingsanti-CD3 mAb soluton whose final concentration was 1ug/ml was added to cell culture media of above groups. 6 hours later,The proliferated CD4~+ T lympho- cytes suspension were divided averagely at random into 6 groups:(1), Group blank contrast:no medicine was added to cell culture medium;(2), Group CsA.CsA soluton whose final concentration was 50ng/ml was added to cell culture media;(3), Group low-concentration SIN: SIN soluton whose final concentration was 10 umol/L was added to cell culture media;(4) , Group middle-concentration SIN: SIN soluton whose final concentration was 200 umol/L was added to cell culture media;(5), Group high-concentration SIN :SIN soluton whose final...