Cases Review And Experimental Research On Interference With Muscle Cell Damage Of Jianpi-qushi Decotion On Myasthenia Gravis

Purposes:Myasthenia gravis is a synapse dysfunction disease between acetylcholine motorneuron and skeletal muscles.Its pathogenesy is due to acetylcholine antibodies which lead to the transmission block of neuromuscular junction. But nowaday some researches also discovered that there was muscle cell damage in myasthenia gravis,like paramorphia in mitochondria,missing of respiratory comples 1 et cetera,which could not simply be explained by the transmission block of neuromuscular junction.On the other side,my tutor-professor Liu Youzhang,applied Jianpi-qushi(JQ) decotion to treat myasthenia gravis and achieved satisfactory curative effect.Therefore this project is to carry out clinical retrospective investigation,to summarize the current situation of diagnoses and treatment of myasthenia gravis,to observe the effect of strengthening spleen and benefiting qi method on myasthenia gravis,and to research the pathology and pathogenesy of muscle cell damage in myasthenia gravis,filling up the pathogenesy of myasthenia gravis,to observe the effect of JQ decotion and discuss its curative effect on muscle cell damage of myasthenia gravis,to provide theory guidance for the combined treatment of Chinese medicine and western medicine.Methods:This project includes documentary research and experimental research.In documentary research,documents of myasthenia gravis were sorted and summarized in detail at four aspects,like knowledge of myasthenia gravis in Chinese medicine,researches on myasthenia gravis in mordern medicine,the relationship between myasthenia and muscle cell damage,researches on the models of myasthenia gravis.Experimental research consist of two parts.The first part is cases review on myasthenia gravis.The cases of myasthenia gravis in the near 10 years from the medical records department of the first affiliated hospital of Guangzhou University of Chinese medicine were to refered to.To analyse the causation,pathogenesis,symtoms,inducement,complication and treatment of myasthenia gravis.Then statistic analyses were to carry out. The curative effect of drugs of strengthening spleen and benefiting qi were to observed.The current situation of diagnoses and treatment of myasthenia gravis was to summarized.The second part was animal experimental research. 45 specific pathogen-free Lewis rats were chosen.6 of them were randomly selected into normal group(group A),which were raised normally.The rest of the rats were used to produce models of experimental autoimmune myasthenia gravis(EAMG) via subcutaneous injections of man-made synthetical peptide of acetylcholine receptorαsubunit 97～116.6 weeks later,36rats left.32 EAMG models were confirmed via electromyography,which were randomly divided into model control group(group B),prednisone group(group C),JQ decoction high dose group(group D),JQ decoction low dose group(group E).Each group contained 8 EAMG rats.2 weeks after EAMG was confirmed,drugs interference via intragastric administration began.Group C was given prednisone 5.4mg/kg body weight(BW)·d.Group D and E was given Jianpi-qingre-qushi decoction respective in the dose of 37.6mg/kg BW·d,18.8mg/kg BW·d.Group A and B were given the same amount of physiological saline.The amount of intragastric administration is 10ml/Kg BW,which was given once per day for 4 weeks.At the end of the experiment,general conditions,Lennon grade,ultrastructure of the muscle mitochondria and neuromuscular junction(NMJ) of the rats were observed. The DNA glycosylase expression,the amount of ATP,and the active and content of cytochrome oxidase(CCO) of the muscle mitochondria were detected.Results:1.Review on clinical study:1.1 In in-patient of myasthenia gravis,the male female ratio was 1:1.23.The morbidity peak lied between age 31 to 50.72.73%of the patients' course were below 5 years.Most of them belonged to midrange generalized type.1.2 Active infection prevention and control,sufficient rest,avoidance of exertion,regular drug administration,never change drug dose personally,healthy diet,and agreeable emotion played a very important role in the control and recover of the patients' condition.1.3 Most of the tongue and pulse presentations of myasthenia gravis were light tongue,white fur,and thready pulse,which reflected the main pathogenesis of myasthenia gravis that is spleen and kidney deficiency with turbid damp inside.1.4 The analyse of the frequency of Chinese drugs used in the treatment of myasthenia gravis showed:the most commonly used drugs was drugs that can invigorating qi and strengthening spleen.Other drugs included drugs invigorating kidney,drugs eliminating dampness,drugs activating blood circulation etc.This result accorded with the analyse of the main pathogenesis of myasthenis gravis.2.Animal experimental research:2.1 JQ decoction could improve the general conditions of EAMG rats,including symtoms,color and luster of their fur.The difference of BW before and after the administration was not significant between group B and C(P＞0.05).The BW of group D and E increased significantly after administration(P＜0.05),but the difference between group D and E was not significant(P＞0.05).2.2 Electron microscope observation of muscle mitochondria and neuromuscular junction:In EAMG rats,there were sparse mussy muscle fibre,decreasing mitochondria. And part of the winkle in NMJ were mussy,diffluent,disappeared,and uesicae in presynaptic membrane decreased,vacuolization in organelles like mitochondira existed.After administration of JQ decoction,the pathology changes mentioned above were improved,which showed JQ decoction could improve the MCD of EAMG.2.3 The expression of OGG1,NTH1,MPG of muscle mitochondria in group A was the lowest,and those in group B is very high.The difference between group A and B was significant(P＜0.01).The expression in group C decreased significantly than group B(P＜0.01),but still higher than group A(P＜0.01).The expression in group D and E were significantly higher than the other three groups(P＜0.01),but there was no significant difference between group D and E.2.4 The content of ATP of muscle mitochondria in group B decreased, significantly different from that in group A(P＜0.01).After the administration, the content in group C,D and E all increased,group D and E were higher than group C(P＜0.05),and group D were higher than group E(P＜0.01).2.5 The activity and content of CCO of muscle mitochondria in group B decreased, significantly different from that in group A(P＜0.01).After the administration, the activity and content in group C,D and E all increased.Group D and E were higher than group C(P＜0.05),and group D were higher than group E(P＜0.01).Conclusion:1.EAMG rat models could be established via subcutaneous injections of man-made synthetical peptide of acetylcholine receptorαsubunit 97～116.2.JQ decoction could improve the general Conditions of EAMG rats,including symtoms,color and luster of their fur,and their BW.3.JQ decoction could improve the pathology changes in muscle mitochondria and NMJ of the EAMG rats.4.JQ decoction could significantly improve the expression of OGG1,NTH1,MPG of muscle mitochondria in EAMG rats.5.JQ decoction could significantly improve the amount of ATP in muscle mitochondria of the EAMG rats.6.JQ decoction could significantly improve the activity and amount of CCO in muscle mitochondria of the EAMG rats.