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The Effect of FTY720on the Expression of MMP-9in the Brain Tissue Experimental Autoimmune Encephalomyelitis Mice

Author: TanShuWei
Tutor: YuJuMing
School: North Sichuan Medical College
Course: Neurology
Keywords: experimental autoimmune encephalomyelitis (EAE) multiple sclerosis MMP-9 FTY720
CLC: R744.3
Type: Master's thesis
Year: 2014
Downloads: 1
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Abstract


Objective: To observe effect of FTY720on the expression of MMP-9inthe brain tissue of EAE, and compare the expression changes with differentdoses of FTY720.Methods: The animal model was established in female C57BL/6miceby immunizing mice with myelin oligodendrocyte glycoprotein35-55(MOG35-55),complete freund’s adguvant (CFA) and pertussis toxin (PTX).One hundred twenty C57BL/6mice were randomly divided into six groupsequally: normal control group,CFA group,EAE group,different doses FTY720groups (10mg/kg intervention group,3mg/kg intervention group,1mg/kgintervention group).Mice were fed with FTY720in different doses FTY720groups,starting from the onset of the disease to the day mice were sacrificed.Clinical signs of mice were assessed twice a day by two investigators. Micewere examined daily for clinical signs and scored on a scale0-5according tothe signs and symptoms. Hematoxylin-eosin staining and Luxol Fast Bluemyelin staining were performed on the sections of mice’brain.The number ofinflammation focus in the brain tissue was counted under the opticalmicroscope. The expressions of MMP-9in brain tissue were detected byimmunohistochemistry technique, while using real-time PCR technology toobserve the effect of drug intervention on mRNA expression of MMP-9inmice. Experimental results were analyzed with SPSS17.0.Results:1.No disease phenotype was observed in normal control group and CFAgroup.Comparing to EAE group,1mg/kg intervention group had degraded in neurological deficitscores,weight1oss and eclipse period(P<0.05),but had nodifference inmorbidity;while10mg/kg intervention group and3mg/kgintervention group’s mice have decreased neurological deficit scores,weight1oss,eclipse period and morbidity (P<0.05).Comparing to1mg/kgintervention group,10mg/kg intervention group and3mg/kg interventiongroup had degraded inneurological deficit scores,weight1oss and eclipseperiod (P<0.05).2.The brain tissue’s hematoxylin-eosin (HE) staining histopathologyresults of EAE group’s mice indicated that brain tissue already hadinflammation while no change in normal control group and CFAgroup.Comparing to EAE group,the extent and amount of the inflammationfocus decreased in the10mg/kg intervention group,3mg/kg interventiongroup and1mg/kg intervention group(P<0.05).Comparing to1mg/kgintervention group,the extent and amount of the inflammation focus decreasedin the10mg/kg intervention group and3mg/kg intervention group(P<0.05).Luxol fast bule-cresyl fast violet myelin staining indicated nodemyelination lesions in normal control group and CFA group.In EAE groupthe demyelination lesions were the most serious.Dispersive demyelinationlesions could be seen casually in the10mg/kg intervention group, and3mg/kgintervention group and1mg/kg intervention group.3.Immunohistochemistry and image analysis results of each group miceindicated that the brain tissue expression level of MMP-9had increased in theEAE group comparing with normal control group and CFAgroup.(p<0.05).Comparing to EAE group,10mg/kg intervention group,3mg/kg intervention group and1mg/kg intervention group’s mice had lowerlevel of MMP-9(P<0.05).Comparing to1mg/kg intervention group,10mg/kg intervention group, and3mg/kg intervention group’s expression level ofMMP-9have decreased (p<0.05).4.Quantitative PCR detection results of each group mice indicated thatthe brain tissue expression level of MMP-9mRNA had increased in the EAEgroup comparing with normal control group and CFA group.(p<0.05).Comparing to EAE group,10mg/kg intervention group,3mg/kg interventiongroup and1mg/kg intervention group’s mice had lower level of MMP-9mRNA (P<0.05).Comparing to1mg/kg intervention group,10mg/kgintervention group, and3mg/kg intervention group’s expression level ofMMP-9mRNA have decreased (p<0.05).5. FTY720intervention dosage and neurological score was negativelycorrelated(r=-0.779, p=0.000);FTY720intervention dosage and MMP-9protein expression was negatively correlated (r=-0.759, p=0.000),FTY720intervention dosage and MMP-9mRNA expression was negatively correlated(r=-0.801, p=0.000)。All were statistically significant.Conclusion:1.FTY720ameliorates EAE mice and the effects are probablly doseagedependence.2.One of the mechanisms related to FTY720’s amelioration on EAE is toinhibit the expression level of MMP-9protein and mRNA in the braintissue,which help to maintain the integrity of blood brain barrier.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Spinal cord disease > Myelitis
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