Measurement Of Related Cytokines In Low Density Lipoprotein Receptor-related Protein 4 Induced Myasthenia Gravis In Mice

Objective:To establish an experimental autoimmune myasthenia gravis(EAMG)model in mice to study the pathogenicity of low density lipoprotein receptor related protein 4(LRP4)in myasthenia gravis(MG)and its relationship with related cytokines.Methods:10 female C57bl/6 mice of 8 weeks old were randomly divided into experimental group(n=5)and control group(n=5),which weighted from 18-25g.The mouse EAMG models were made by using the method of active immunization,subcutaneous injection of LRP4,and gastrocnemius frozen sections prepared,The sections were treated with tetramethylrhodamine-alpha-bungarotoxin(TR-BTX)to localize acetylcholine receptor(AChR,red),and FITC conjugated goat anti-mouse IgG antibody to label anti-LRP4 antibody(green),and observed under laser confocal scanning microscope(LCSM).The concentrations of cytokine IL-12,IFN-y and IL-17 in sera of two groups of mice were measured by ELISA kit.Results:In the experimental group,some mice showed slow motion.The ability of biting and grasping was weakened and the weights slightly decreased.One mouse had hair removal around the left eye and could not open the eye.The surface of the gastrocnemius showed green fluorescence and red fluorescence in the same location,indicating that the LRP4 antibody had bounded to LRP4 antigen in neuromuscular junction(NMJ).The concentration of cytokine IL-12,IFN-y and IL-17 increased in the experimental group,but there was no significant difference(p>0.05)by the spss17.0 statistical analysis.Conclusion:It is preliminarily proved that LRP4 can induce EAMG in C57BL/6 mice,and LRP4 antibody is one of the pathogenic antibodies of EAMG.It can not directly explain the correlation between IL-12,IFN-y,IL-17 and LRP4-MG.