| Objective:This study aims to establish a lentivirus-mediated gene overexpression Fgl2ratmodel of autoimmune myocarditis,which will reveal the effect on heart function andrelated inflammatory cytokines in autoimmune myocarditis rat; To clarify itstherapeutic effect, looking for effective autoimmune injury therapeutic targets.Methods:Fgl2lentiviral overexpression autoimmune myocarditis group (n=8), Fgl2overexpression lentivirus vector5x107IU/n, diluted to1ml saline bolus tail vein.②Empty vector GFP autoimmune myocarditis group (n=8), GFP lentiviral vector5x107IU/n, diluted to1ml saline bolus tail vein.③Autoimmune myocarditis group(n=8).④Normal group(n=8). Natural light, normal diet, free access to water.In60days,testing LVEF, FS, and HR by ultrasonic,then take out the blood, detected theBNP、IL-17、IL-6and TNF–alpha. Removing the heart and clearing it by NS.Cut offatrial and ventricular in horizontal incision, fixing the atrial in formalin, biopsy,andobserved the athological by HE. ventricular part was frozen sections,then watchingthe fluorescent virus ransfection in fluorescence microscope. The expression of Fgl2was tested by werstern blot.Results:(1)In autoimmune myocarditis group,the activity and the amount of drinkingwater and food were reduced.(2)Fluorescence detection:in EAM-Fgl2group,the green fluorescence can beobserved in myocardial tissue.(3)Analysis the mRNA expression of Fgl2by quantitative PCR(4)Western blot detected the target portein: compared with the normal group,the target protein expression of Fgl2in EAM-FGL2group was increased,while thereis no significant difference in EAM and GFP EAM. compared with the EAM group,the related protein expression of Fgl2in EAM-FGL2group was increased. (5)HE staining of myocardial:normal group, myocardial cells arranged in neat,nucleus are the same size, cytoplasmic staining is uniform,myocardial fibersarranged in neat.EAM group, there are small focal necrosis in subendocardial. Fgl2overexpression EAM group,it can be seen that a large area of necrosis appear in leftand right ventricular epicardial, room mediastinal has extensive myocardial necrosis,Fibroblast proliferation and observed a large number of neutrophils, macrophagesacidic cell.(6)Compared with the normal group, the LVEF and FS of the EAM-Fgl2groupis significantly lower.The difference between two groups has statisticalsignificance(P<0.05). Compared with autoimmune myocarditis group, the LVEF andFS of the EAM-Fgl2group are significantly lower.The difference between twogroups has statistical significance(P<0.05). Compared with EAM group, the LVEFand FS of the EAM-GFP are the same,and the difference between two groups has nostatistical significance((P>0.05).Conclusion:(1)Fgl2overexpression vector can significantly increase the expression of Fgl2gene in EAM group.(2)The expression of Fgl2gene can deteriorate the cardiac function of EAM rats;(3) The overexpression of Fgl2can upregulated the level of related inflammatorycytokines which lead to that the immune response towards the progress of immuneinjury. |
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