| Background:Multiple sclerosis is an autoimmune disease which matters demyelinating lesions as the main pathological features in the central nervous system.Generally,the average oneset age of MS is between 20 and 40,most of them are females.The clinical manifestations included as follows:Neurological deficits and sensory dysfunction such as Visual impairment,rotating type nystagmus,weakness of one or more body,abnormal sensation as pain,numbness which incidents facial or other parts of the body,different degrees of ataxia and so on;Autonomic nerve dysfunction such as frequent urination,urinary incontinence,constipation,diarrhea,sweating;psychiatric symptoms include depression and irritable;Memory loss,poor judgment which belong to cognition impairment.Multiple symptoms and signs of space and time appear repeatedly in the course of the multiple is the main clinical characteristics of MS.Most patients presented with recurrent nerve dysfunction,the disease develops more and more serious,cause significant impact on the quality of life of the individual,and to bring huge economic burden to the family.At present,the treatment of multiple sclerosis and more concentrated in the inhibition of the body's immune system,can reduce the brain's inflammatory injury.But with the disease continues to increase,the gradual increase in drug dose,these drugs inhibit the body's immune system also brings significant side effects to the human body.The application of traditional Chinese medicine can effectively improve such a deficiency,through the combination of traditional Chinese medicine to a certain extent,can reduce the dose of immunosuppressive agents,reduce side effects.At the same time,traditional Chinese medicine can also reduce the clinical symptoms of multiple sclerosis,reduce the number of recurrence.Purpose:After different drugs act on EAE rats,observing changes in rats nerve function score,weight change,conventional HE staining the EAE rats multiple levels of the brain,spinal cord pathology,the determination of different groups in the rat serum after intervention of CD4~+T cell surface expression of CD28,CTLA4 quantity.For exploring traditional Chinese medicine in the acute phase of intervention possible mechanism of the influence on multiple sclerosis,and providing a certain basis that combining traditional Chinese and western medicine in the treatment of multiple sclerosis has more advantages than pure western medicine or Chinese medicine therapy.Methods:(1)Experimental animal grouping and administration:set up traditional Chinese medicine(Bu Shen Jie Du Tong Luo)group,set the hormone group and blank group,EAE group and combining traditional Chinese and western medicine group.(2)EAE model:guinea pig spinal cord homogenate production of antigen,immune-induced SD rats acute EAE model,rats were observed for clinical signs,record daily weight and neurological score.(3)Administration:From the day of the immune,the Chinese medicine group was 5g/kg dose for traditional Chinese medicine(TCM)1 ml lavage;Dose hormone group 1mg/kg per day to give the prednisone acetate dragon suspension mixed liquid of 1.25mg/ml,1 ml lavage;Hormone combined with Chinese traditional medicine group were given Chinese medicine every day 1 ml and 1 ml of 1.25 mg/ml prednisone acetate lavage,dragon suspension mixed liquid lavage 17 days in a row.(4)Sample collection,observed the pathological and measurement indicators:When it is 18s day after immune,Tail cutting method is used to collect the rat peripheral blood,Then using flow cytometry to detect the quantity of the expression of CD28 and CTLA 4 which are on CD4~+T cell surface in peripheral blood.Saline infusion and ice heart,take off the brain tissue and Lumbar intumescence of EAE rats,after a fixed 4%paraformaldehyde solution,paraffin embedding,Histological section,With the method of HE staining to observe the pathological changes.Results:(1)Rat incidence and body weight change:the blank group without the disease,the incidences of traditional Chinese medicine group,the hormone group and combining traditional Chinese and western medicine group rats compared with the model group had statistical significance(P<0.05),the difference of weigh in different groups compared were meaningless(P>0.05)before the rats had been immune.After accepting the immune the rats appeared not to diet,to restrict movement phenomenon,since then the weight of rats increased briefly,prior to the onset of fall again,blank group rats weight was rising all the way,the weight of EAE group and medicinal group rats increased briefly,prior to the onset of fall again,Blank group rats weight is rise all the way,the weight of Model group,Chinese medicine,Chinese and western medicine group were significantly lower than the blank group(P<0.01).The difference of weigh between traditional Chinese medicine group and traditional Chinese and western medicine group(P>0.05)were meaningless.The difference of weigh between western Chinese medicine group and EAE group(P>0.05)were meaningless.(2)The comparison of incubation period and nerve function score:EAE group attacked after 7 days when rats were immune,traditional Chinese medicine group attacked after8 days when rats were immune,western medicine group attacked after 7 days when rats were immune,Chinese and western medicine group after 9 days when rats were immune.The incubation period of Traditional Chinese medicine with Chinese and western extender than the model group(P<0.05).The western medicine group compared with model group had a lengthen the incubation period,but no significant difference(P>0.05).nerve function score:the nerve function scores of traditional Chinese medicine group,the hormone group and combining traditional Chinese and western medicine group rats compared with the model group had statistical significance(P<0.05)after immuned 7days,There are not significant differences among traditional Chinese medicine,western medicine group,Chinese and western medicine group in terms of statistical(P>0.05).(3)Morbidity peak inflammatory lesions:the blank group without the inflammatory lesions,compared with model group,traditional Chinese medicine,Chinese medicine hormone group,hormone group were significantly reduced(P<0.01),but traditional Chinese medicine group had more inflammatory lesions than hormone group Chinese medicine hormone group(P<0.05),no significant differences between traditional Chinese medicine group and hormone group(P>0.05).(4)CD28,CTLA-4 expression quantity which are on the surface of CD4~+T cell in peripheral blood during the peak period of incidence:CD28:model group compared with normal group was obviously increased(P<0.01),the expression of traditional Chinese medicine,Chinese medicine hormone group,hormone group lower than the it expressed on the model group(P<0.05).No significant differences between traditional Chinese medicine hormone group and hormone group(P>0.05).CTLA-4:model group compared with normal group was obviously less(P<0.01),he expression of traditional Chinese medicine,Chinese medicine hormone group,hormone group higher than the it expressed on the model group(P<0.05).No significant differences between traditional Chinese medicine hormone group and hormone group(P>0.05).Conclusion:part of the mechanism of Bushen Jiedu Tongluo Fang for the role of MS through reducing the expression of CD28 and increasing the expression of CTLA-4 which both exist on the surface of CD4~+T to inhibit excessive immune response.To adjust the EAE rats immune imbalance,relieve clinical symptoms. |
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