Mechanism Of Moderate Ethanol Drinking-inhibited The Systemic Lupus Erythematosus Progression

Background and Aims:Systemic lupus erythematosus(SLE)is a chronic autoimmune disease characterized by high level of autoantibodies and multi-organ tissue damage.It has been indicated that moderate ethanol drinking has a protective effect against the development of SLE by meta-analysis,but there are still no such cases to prove the point through experimental methods so far.Our study is aim to use the experiment methods to prove moderate ethanol drinking delays the SLE disease progression and to explore its functional mechanism.Methods:1,Lupus-prone mice MRL/lpr were treated with 10%(v/v)ethanol or pure water,and we used histopathological H&E stain to observe the severity of skin,joint;used the reagent strips to measure the proteinuria in these two group mice and observed the mortality of the two groups.2,To investigate whether ethanol affects the function of SLE activated T cells,we used anti-CD3 plus CD28 antibodies to conduct normal C57BL/6(B6)mouse T cells to establish the model of activated T cells.We used light microscope and CCK-8 kit to observe T cell aggregation and proliferation,used flow cytometry assay to observe T cell adhesion molecules and used ELISA method to investigate the effect of ethanol on the T cell secreted cytokine;To explore the effect of ethanol on T cell lipid rafts,we used immunofluorescent method to observe the T cell lipid raft clustering after ethanol treating;To investigate whether ethanol affects the function of T cell by regulating lipid rafts,we used lipid rafts scavenger M?CD to conduct T cells.3,In order to investigate whether ethanol relieves lupus serum-induced skin inflammation by regulating lipid rafts,B6 mice were intradermally injected with lupus serum after drinking 10%ethanol(systematic experiment)or B6 mice were injected with ethanol plus lupus serum(local experiment),then we used H&E stain to evaluate the severity of skin in each group.We used light microscope and flow cytometry method to observe whether ethanol or M?CD inhibits lupus serum-induced monocyte differentiation into dendritic cells(DCs).4,To investigate whether ethanol we used has side effect on mice organ tissue and cells,we used ALT kit,H&E stain and CCK-8 kit to observe the effects of ethanol on mice organ tissue and cell viability.Results:1,In the preventive experiment,moderate ethanol drinking relieved the severity of skin and joint in MRL/lpr mice,reduced the level of proteinuria and mortality;in the therapeutic experiment,moderate ethanol drinking reduced the mortality of MRL/lpr mice,but it had no obvious effect on the severity and incidence of skin or the level of proteinuria.2,Ethanol treatment in vitro inhibited the aggregation and proliferation of activated T cells,down-regulated the expression of T cell CD44 and the level of IFN-?,but up-regulated the expression of L-selectin CD62L;Ethanol treatment reduced the percentage of T cell lipid raft clustering and the MFI of lipid raft marker GM1;Lipid raft scavenger M(3CD affected the aggregation,the proliferation of T cells and reduced the percentage of T cell lipid raft clustering.3,In the systemic experiment,drinking ethanol for 2 weeks had no significant alleviative effect on lupus serum-induced skin inflammation,but in the local experiment,ethanol relieved the lupus serum-induced skin inflammation obviously;Ethanol treatment in vitro inhibited the process of lupus serum-induced monocyte differentiation into DCs and reduced the TNFa level which produced in this induction process;Lipid raft scavenger M?CD inhibited lupus serum-induced monocyte differentiation into DCs.4,Ethanol treatment in vivo had no significant effect on the liver ALT and organ tissue,the concentration of ethanol treatment in vitro didn't influence cell viability.Conclusions:1,Moderate ethanol drinking delays the disease progression of SLE;2,Moderate ethanol affects the function of activated T cells by inhibiting lipid raft clustering;3,Moderate ethanol inhibits lupus serum induced monocyte differentiation into DCs and then affects lupus serum induced skin inflammation by regulating lipid rafts.