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Effects of Red Clover Extract on Murine T Lymphocyte Behaviors and Macrophage Behaviors and Skin Allograft

Author: YangZhi
Tutor: ZengYaoYing
School: Jinan University
Course: Immunology
Keywords: RCE T lymphocytes activation proliferateion Phagocytosis NO skin allograft
CLC: R285
Type: Master's thesis
Year: 2009
Downloads: 31
Quote: 0
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Abstract


Objective:To investigate the effects of red clover extract（RCE） on activation and proliferation of murine T lymphocyte and NO secretion and phagocytosis of macrophage and skin allograft,elucidate its underlying mechanisms with immunocell,in order to develop its potential effect in immune suppression.Methods:The cytotoxic effect of RCE on cell suspension was estimated by MTT test.With the stimulation of Con A or PDB,multi-fluorescence staining plus flow cytometrics were used to detect the activation of murine T lymphocyte in vitro;With the stimulation of Con A,CFDA-SE staining plus flow cytometrics to analyze T lymphocyte proliferation in vitro;Fluo-4/AM staining plus flow cytometrics were used to detect[Ca2+]i of lymphocytes of mouse in vitro;The effect of RCE on NO secrete of murine macrophage and RAW264.7 cell line was detected by Griess reagent system in vitro;1μm & 2μm fluorescence beads plus flow cytometrics to analyze phagocytosis of murine macrophage and RAW264.7 cell line in vitro; Murine skin allograft model was established,the effect of RCE on skin allograft was evaluated,mixed lymphocyte reaction was measured.Results:1、RCE have little side-effect on murine lynlphocyte,murine macrophage and RAW264.7 cell line in vitro. 2、With the stimulation of Con A,RCE significantly downregulated expression level of CD69,CD25,CD71 of murine CD3+T lymphocyte in a dose-dependent manner（P＜0.01）;With the stimulation of PDB,RCE inhibited the expression of CD69 and CD25 of murine CD3+T lymphocyte notely in a dose-dependent manner （P＜0.01）;With the stimulation of Con A,RCE inhibited the proliferation of murine T lymphocyte in a dose-dependent manner（P＜0.01）;RCE can promote the Ca2+ inflow of lymphocytes of mouse（P＜0.01） in vitro.3、With the stimulation of LPS,RCE inhibited the NO secretion and phagocytosis of murine macrophage and RAW264.7 cell line notely in a dose-dependent manner （P＜0.01） in vitro.4、The immunosuppressive properties of RCE effectively promoted allograft skin survival time in the murine skin allograft model.Besides,RCE could cooperate with CsA.Conclusion:The immunosuppressive properties of RCE on murine T lymphocyte mainly display on the inhibition of CD69,CD25,CD71 expression and proliferation in vitro. Combine with the inhibition of RCE on NO secretion and phagocytosis of murine macrophage and RAW264.7 cell line,we found that RCE had potential effect on antiinflammatory.These immunosuppressive properties of RCE could promote allograft skin survival state,and the immunosuppressive effect of RCE could also associated with CsA,these data provide useful information in which RCE could develop a new immunointervention reagent.

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