Study On The Effect Factors About C57BL/6 Mice Modeling The Experimental Autoimmune Encephalomyelitis And Therapy By Rapamycin

Part One The study on the effect factors about C57BL/6 mice modeling theexperimental autoimmune encephalomyelitis(EAE)Objective To investigate the influence of different effect factors such as the concentration difference of myelin oligodendrocyte glycoprotein35-55(MOG35-55)or Pertussis toxin(PTX)on the success rate and mortality rate of C57BL/6 mice modeling the experimental autoimmune encephalomyelitis(EAE).Methods 1.Experimental materials:30 female C57BL/6 mice,aged 6weeks,weight 16-18 g,bought from the Animal Reach Center of Gansu University of Chinese Medicine(Gansu,Lanzhou).The mice were feeded in Animal Experiment Apartment of PLA Lanzhou General Hospital's SPF Animal Lab,keeping the indoor temperature between 22-26? and 12 h circulation between dark and light,supplying enough food and clean water.All the animal processes were allowed by the Animal Experimental Ethics Committee of the Second Hospital of Lanzhou University.2.EAE model induce and effect factors control:Divide the 50 mice into groups include blank group,control group,MOG group and PTX group randomly.Mix and emulsify the MOG35-55 and Freund's adjuvant complete(CFA)then inject different concentration into four positions on both sides of the spine subcutaneously.Meanwhile on day 1 and day 3 during MOG35-55 immunization,PTX was injected intraperitoneally as plan.At least 2 investigators weighed and evaluated the animals for clinical score in a blinded way every day.The obtained tissue was presented to the pathological pathology department of the Second Hospital of Lanzhou University to do the HE-staining.Result 1.The mice of blank team survived well for their weight increased and keep around20-21g;the mice of the control team firstly showed weight decreased in the 7th day;the first morbidity happened in the 7th day and increased to the peak in the 17 th day,whose morbidity rate is 100% while the death rate is 20%.The mice of the MOG team firstly showed weight decreased in the 5th day;the first morbidity happened in the 6th day and increased to the peak in the 17 th day,whose morbidity rate is 100% while the death rate is 0.The mice of the PTX team firstly showed weight decreased in the 10 th day;the first morbidity happened in the 10 th day and increased to the peak in the 17 th day,whose morbidity rate is 100% while the death rate is 0.2.As the results of statistical analysis shows: the weight changes and clinical scores among the control team,MOG team and PTX team is significant(P<0.01),but nonsignificant between every two teams.3.The brain and spinal cord tissue were extensively infiltrated of inflammatory cells showing in the HE-staining.Part Two The comparative study between rapamycin and methylprednisolone used totreat C57BL/6 experimental autoimmune encephalomyelitis(EAE)model miceObjective To observer the different effect between rapamycin and methylprednisolone sodium succinate used to treat C57BL/6 experimental autoimmune encephalomyelitis(EAE)model mice.Methods Obtain 15 modeled EAE mice whose clinic score between 2.5-4.0 from our lab,then divided them into control group,rapamycin group and methylprednisolone group injected the normal saline,rapamycin and methylprednisolone sodium succinate for 7 days.At least 2investigators weighed and evaluated the animals for clinical score in a blinded way every day.The obtained tissue was presented to the pathological pathology department of the Second Hospital of Lanzhou University to do the HE-staining.Results 1.Without infecting drugs,the weight of EAE team's mice showed slightly decreased but keep up and down around 16 g while whose clinical scores increased meaning the state of the illness became worse so 2 mice died during this process.After infecting MP,the weight of these mice increased since 2nd day while whose clinical scores became lower and lower meaning the state of the illness changed better but 1 mouse still died.After infecting rapamycin,the weight of these mice did not change any more but their clinical scores suddenly increased in the 4th day meaning the state of the illness became worse and 1 mouse died.2.As the results of statistical analysis shows: the weight changes and clinical scores among the EAE team,MP team and RAPA team is nonsignificant(P>0.01),the clinical scores between EAE team is significant.3.With the disease moving on,the EAE group and RAPA group show worse damage in nerve cell but better change,and RAPA can't alleviate the infiltrated of glial cells showing in the HE-staining.