The Immunotoxicity Effect Of Aluminum Chloride On Splenic Lymphocyte Of Chichen In Vitro

Aluminum trichloride(AlCl₃) of different concentration was added to culture solution of lymphocyte from chicken splen.The identical volume culture solution was added to control group. The cytoactive,cell proliferation,the cytokine,the anti-oxidations,the calcium homeostasis and cell apoptosis of lymphocyte cultured in vitro were detected by MTT colorimetric, radioimmunoassay,RT-PCR and flow cytometry to reveal the immune cytotoxicity effect of AlCl₃ on chicken and provide basal data for the relief of aluminum toxicity.The results showed as follows:1 The IC₅₀ of AlCl₃ to chicken splenic lymphocytes detected by trypan blue and MTT was 0.832±0.01 mg/ml cultured for 24 hours in vitro.2 The low dose aluminum inhibited the proliferation of T cell,but did not inhibit that of B cell.The high dose aluminum inhibited both T and B cell proliferation.3 Aluminum increased the leakage of lactate dehydrogenase and decreased cytoactive of lymphocyte.The results indicated that aluminum changed the membrane permeability of lymphocyte.4 The contents of IL-2 and TNF-αof lymphocyte culture fluid in the trial groups were significantly lower than those of in the control group(p＜0.01;p＜0.05),and there were negative correlation between aluminum dose and their contents.The results indicated that the decreased contents of IL-2 and TNF-αwere one of the immunotoxicity mechanisms of aluminum.5 The activity of nitricoxide synthase(NOS),the contents of nitrogen monoxidum(NO) and malonaldehyde(MDA) of lymphocytes were higher than and the activity of hepatocuprein(SOD) and glutathione peroxidase(GSH-Px) were lower in the trial groups than those of in the control group(p＜0.01;p＜0.05) and there were dose-effect correlation with AlCl₃.The results indicated that aluminum induced oxadation stress to splenic lymphocytes of chicken and resulted in the immunotoxicity effect.6 The splenic lymphocytes apoptosis were observed by flow cytometry and AO/EB double fluorescent staining.Typical apoptosis characters were observed by fluorescence microscope.The cell number of G₀/G₁ was more and cell number of S and G₂/M were fewer,the apoptosis rate and apoptosis index were higher in the trial groups than those of in the control group (p＜0.01;p＜0.05) and there were dose-effect correlation with AlCl₃.The results indicated that aluminum caused immunotoxicity effects by inducing chicken immunocyte apoptosis.7[Ca²⁺]i was significantly higher and the level of CaMmRNA was significantly lower in lymphocyte of trial groups than those of in the control group(p＜0.01;p＜0.05).The results indicated that AlCl₃ caused the disequilibrium of calcium homeostasis.