Effects Of Alkaloid Sinomenine On The Activity Of ERK1/2 And Proliferation Of CD4～+T Lymphocytes Of Human Peripheral Blood In Vitro

Object: To observe the Effects of alkaloid sinomenine on the activity of ERK1/2 of Ras-MAPK signal transduction pathway and proliferation of CD4~+T lymphocytes of human peripheral blood in vitro for study of the efficacy and molecular biological mechanism of SIN immunosuppressive human CD4~+T lymphocytes. This research aims at providing theory foundation to clinical make use of SIN in the field of transplantation, and providing a safe, effective and inexpensive administration scheme that can be offered to immunosuppressive drug therapy for recipients.Methods: Establishment of human CD4~+T lymphocyte models of peripheral blood:1,50ml elbow vein blood in three healthy adult male volunteer was draw, adding 500U heprin to anticoagulation; PBMC were isolated from the vein blood with lymphocytes separation medium;CD4~+T lymphocytes wereisolated from PBMC suspension with immunomagnetic beads and cultured(37℃, 5%C02) with 10%FCS RPMI-1640;The CD4~+T lymphocytes were proliferated by PHA(final concentration: 5ug/ml) and IL-2 (final concentration:100 U/ml) for 7 days. The proliferated CD4~+ T lymphocytes suspension of 3 persons were treated as followings:1) SIN of different concentration effected on cell culture medium : (1)Group blank contrast: no medicine was added to cell culture medium; (2)Group low-concentration SIN: SIN solutrope whose final concentration was 10 umol/L was added to cell culture media;(3)Group middle-concentration SIN:SIN solutrope whose final concentration was 200 umol/L was added to cell culture media; (4), Group high-concentration SIN :SIN solutrope whose final concentration was 1000 umol/L was added to cell culture media;(5)Group CsA:CsA soluton whose...