Anti-inflammatory Effects Of Rapamycin On Experimental Autoimmune Encephalomyelitis In Mice

Objective: In this experiment,EAE mice were used as animal models,and Rapamycin(Rapamycin,Rapa)was used to intervene.Weaver’s 15-point clinical symptom score was used to score the neural function of EAE mice to assess the incidence of mice in each group.HE staining was used to study the effect of Rapa on the infiltration of inflammatory cells in the spinal cord tissue of EAE mice.At the same time,RT-PCR technology was used to detect the m RNA expression of cytokines: IFN-γ and IL-17 in the spinal cord of mice from the transcription level to analyze the inhibitory effect of Rapa on EAE mice.Through the above experiment to investigate the protective effect of Rapa on EAE mice,so as to guide the dinical use of multiple sclerosis to provide basic experimental basis.Methodology:C57BL/6,Mouse 8-10 weeks,18-20 gconditions in an environment free of special pathogens(SPF).The dark/bright cycle is 12 hours,and regularly supplied mice with rat food and dean drinking water.randomly divided into Rapa,EAE and control groups.EAE and Rapa groups used MOG35-55 as antigens,The animal model was established by complete Freund’s adjuvant(CFA),tuberculin and pertussis toxin,which were injected subcutaneously EAE both sides of the spine.EAE and Rapa set up animal models,control group is not immune.Rapa group started on the day of immunization,Rapa group was given daily doses(1.0 mg/kg)at 8:00 and 16:00 fixed time.EAE and control mice were given the same amount of placebo.On the first day of immunization,In the morning and evening(8:00 and 16:00),the mice were evaluated by two observers.And perform the peak nerve function score,Until death.After 21 days of immunization,the mice were killed and harvested,Take the spinal cord of each group,HE staining;The RT-PCR technique was used to detect the expression of IFN-γ.IL-17 m RNA transcription.Results:1.incidence of mice in each group:according to the incidence of nerve behavior in mice,we used the peak period as a marker,and the difference between the Rapa group and the EAE group was statistically significant(P<0.05).2.HE staining:at 21 days,EAE group and Control group showed that the number of inflamm atory cell infiltration in spinal cord tissue of mice in the EAE group increased significantly,when compared with the EAE group in Rapa group,the number of inflammatory cell infiltration in the spinal cord tissue of mice in the group was significantly reduced.4.RT-PCR:expression of cytokine IFN-γ.IL-17 m RNA in the spinal cord of mice in each group:compared with the EAE group and the Control group,The expression rate of cytokine IFN-γ,IL-17 m RNA in EAE mice increased significantly.And there was a statistical difference(P<0.05),Rapa group compared to EAE group,The proportion IFN-γ,IL-17 m RNA proinflammatory cytokines in the spinal cord of Rapa mice decreased significantly.and statistically significant(P<0.05).Conclusion:1.Rapa reduce the neurological function score of dlinical symptoms.2.Rapa attenuates inflammatory cell infiltration in spinal cord tissues of EAE mice3.Rapa significantly decreased the proportion IFN-γ,IL-17 m RNA proinflammatory cytokines in the spinal cord of mice,and the difference was statistically significant(P<0.05)...