Effects On The Prescription For Invigorating Spleen And Kidney On The Serum IL-4 And TNF-? Of Rats With Experimental Autoimmune Myasthenia Gravis(EAMG)

Objective: Establish EAMG mice models,and by observing the method of spleen kidney to the influence of EAMG mice's weight,Clinical symptoms,AChR-Ab,IL-4 and TNF-?,investigate the mechanism of action of the method of spleen kidney in the treatment of EAMG.Method:Healthy clean female C57 BL / 6 mice,weeks(8-10 weeks),weighing about 18-20 grams,a total of 60,according to the random number table were randomly divided into control group 10 and the experimental model group 50.Fully blend the Mouse Acetylcholine receptor ? subunit advance 97-116 peptide and PBS or CFA according to the 1: 1 volume ratio to prepare a package of anti-oil emulsion mixture.Then in the experimental model group,each mouse shoulders and back foot double foot bottom section(four sites)were given the synthesis of immunogen 50?L respectively,a total of 200?L(murine peptides containing 50?g).Primary immune application contained complete found adjuvant synthesis of immunogenic to carry on the mice immune.On the thirtieth,fiftieth days give contains incomplete found adjuvant synthesis of immunogenic of strengthening immune(configuration method,measuring,injection site with former);blank control group with the same measurement,place,time to give injections of saline solution.Identify the model after 70 days of the first time of immue.During the period of modeling: measure and record the change of the body weight in mice every two days;reference Lennon reclaimed the classification method of clinical symptom rating evaluation test mice;Using enzyme-linked immunoassay detection AChR-Ab drops serum level in mice;Lines of neostigmine injection is administered after fatigue test to observe the mice muscle weakness symptom scores change indicators such as judge EAMG mice model of success;At last,according to the level of only 25 mice can succeed,random divided into normal saline group and 9(0.2 ml),hormone group,prednisone acetate group of 8(9 mg/kg.d),Chinese traditional medicine group,spleen and kidney party group of 8(4.5 g/kg·d),blank control group 10(0.2 ml).By lavage daily dosage or physiological saline respectively,2 weeks of treatment.During symptom scores: record the changes of the groups of mice weight and muscle weakness,before and after treatment were detected by enzyme-linked immunosorbent method in mice serum IL-4,TNF alpha drop degree level.Consequence: According to Inclusion criteria Model,there are finally 25 standard EAMG successful model mice.EAMG mice show significant skeletal muscle fatigue and symptoms of myasthenia gravis,and they eat less,their weight gain slowly and even they lose their weight;the control group of mice have a normal diet,act flexible,without symptoms of myasthenia gravis,and their weight grow steadily.In the aspect of myasthenia gravis clinical symptom scores and serum acetylcholine receptor antibody(AChR-Ab),compared with the control group,the model group show statistically significant differences(P<0.01);after Neostigmine experiments,in the aspect of improving muscle inability,there is a statistically significant difference Before and after administration(P<0.01)in the model mice.After 2 weeks of treatment,compared with the saline group,Jianpibushenfang group and prednisone group show a significantly reduced clinical score,a substantial increase in body weight,decrease in serum AChR-Ab titers,interleukin IL-4 and TNF-?.Compared with the saline group,Jianpibushenfang group and prednisone group show statistically significant difference(P<0.01)changes in body weight and clinical score context.The treatment effect of Jianpibushenfang group was slightly lower than prednisone group,and there was significant difference(P<0.01)between the two groups;compared with the saline group,there is a statistically significant difference(P<0.01)of Jianpibushenfang group and prednisone group in IL-4,TNF-? levels change.In the aspect of IL-4,TNF-? level changes in titer,Jianpibushenfang group is lower than prednisone group.There is a statistical difference(P<0.01)between the two groups.Conclusion:1.This experiment adopts the synthetic immune to the original source of the rat acetylcholine receptor a was the 97-116 peptide segments immune preparation EAMG female susceptible C57BL/6 mice animal models and humans in clinical symptoms,the respect such as AChR-Ab change are similar,this method can be successfully identified as the preparation of EAMG mice.For later research on the pathogenesis of myasthenia gravis(MG)and spleen and kidney of pharmacodynamics provides a preliminary experimental basis.2.Spleen and kidney in the treatment of myasthenia gravis,explore its mechanism might be: By reducing the excessive anomalies in the peripheral blood of IL-4,TNF-? level,and then adjust the abnormal AChR specificity immune response,inhibit AChR-Ab,alleviate nerve muscle membrane damage after a prominent,thus to improve the effect of EAMG laboratory mice symptoms of muscle weakness.