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The Effect of Recombinant Hirudin on the Expression of S1P1Receptor in the Brain Tissue of EAE Mice Model

Author: TanQiang
Tutor: YuJuMing
School: North Sichuan Medical College
Course: Neurology
Keywords: Multiple sclerosis Experimental autoimmuneencephalomyelitis Recombinant hirudin S1P1
CLC: R744.51
Type: Master's thesis
Year: 2014
Downloads: 1
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Abstract


Objective: To investigate the effect of recombinant hirudin on theexpression of S1P1(sphingosine1-phosphate-1) receptor in the brain tissue ofmice with experimental autoimmune encephalomyelitis.Methods:64healthy female C57BL/6mice were randomly divided into3groups, respectively,22mice in the normal control group,22in EAE modelgroup and20in recombinant hirudin treated group.With water in oilemulsion antigen made of complete Freund’s adjuvant and MOG35-55,EAEmodel was established by pertussis toxin. On the fourteenth day HE stainingand LFB staining were used to assess EAE model. On the eighteenth day ofthe intervention group was given recombinant hirudin hypodermic (2.0mg/kg)injection, the normal group and the EAE model group were given the samedose of normal saline by hypodermic injection. Their appetite, mental stateand behavior change were observed daily; meanwhile their weight and neuralfunction defect were scored. Mice were sacrificed at the thirty-second day.The changes of S1P1receptor were observed with immunohistochemistrymethod; detection of S1P1mRNA expression level in brain tissue of mice wasdetected by RT-qPCR.Results:1.The normal group wasn’t found any onset of disease.The recombinanthirudin intervention group was significantly lower than the EAE groupof in nerve function defect score (P<0.05). The neural function defect score ofthe normal group and the recombinant hirudin group showed no statisticalsignificance (P>0.05). 2. Immunohistochemical staining showed expression of S1P1in braincells,in all groups.In normal group and the model group the expression ofS1P1receptor mainly located in the cell membrane, but in recombinanthirudin group in cytoplasm. Between the normal group and the EAE modelgroup, the average optical density difference showed no statisticalsignificance (P>0.05). Comparing with the average optical density either inthe normal group or in the EAE model group, the average optical density inthe recombinant hirudin group was significantly lower (P<0.05).3. RT-qPCR detecting showed the expression of S1P1mRNA in braintissue of mice in the recombinant hirudin group was significantly lower thanthat in the EAE group (P <0.05). S1P1mRNA expression in brain tissues ofmice in EAE group was slightly lower than that in the normal group (P>0.05)Conclusion:1. Recombinant hirudin ameliorates EAE mice.2. One of the therapeutic effects mechanisms of thrombin inhibitorhirudin may lies in inhibiting the expression S1P1receptor by intervening thecoagulation pathway.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Spinal cord disease > Demyelinating disease > Multiple sclerosis
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