| ObjectiveThis study includes clinical research and animal experimental research.From the retrospective study on the clinical cases of myasthenia gravis we want to summarize the correlation between TCM Syndrome Distribution and myasthenia gravis with thymoma and providebasisfor clinical treatment.On the other hand,we selected syndrome of deficiency of spleen qi and stomach qi in patients with myasthenia gravis as the research object,through comparing the difference of PGC-1α mRNA expression in the peripheral blood between myasthenia gravis patients and healthy volunteers to investigate the changes of the expression of PGC-1 alpha mRNA in peripheral blood of MG patients,and explore the role in the pathogenesis of myasthenia gravis.The animal experimental study was to observe the changes of the ultrastructure of skeletal muscle,mitochondria and neuromuscular junction,skeletal muscle PGC-1,NRF-1 and Tfam mRNA expression,and the effect of the Qiang ji jian li formulae through the replication of EAMG rats.We could explore the correlation between experimental indicators,further from the regulation of mitochondrial biosynthesis related pathways to supplement the pathogenesis of myasthenia gravis.We want to clarify the possible mechanism of improving the muscle cell damage and mitochondrial structure and function of EAMG rats,so that the method can provide scientific experimental basis for the treatment of myasthenia gravis with the method of invigorating the spleen and supplementing qi.Methods1.Retrospective case studyThe cases of myasthenia gravis electronic medical records from September 2011 to January 2016 in the first affiliated hospital of Guangzhou University of Chinese medicine.All patients had to meet the diagnostic criteria,inclusion criteria and exclusion criteria.Record the general data of age and sex,clinical type(type I,type II,type II,type III,type IV).According to patients hospitalized for the first time the signs and symptoms,tongue and pulse condition on the dialectical typing,patients could be classified into:deficiency of spleen qi and stomach qi,spleen deficiency and dampness heat,spleen and kidney deficiency.Observation of patients with TCM and Western medicine type associated with thymoma.To investigate the relationship among the TCM syndrome,Western sub-type and thymoma.2.Clinical experimental studyPatients of experimental study are from November 2015 to January 2016 in Digestive department in the First Affiliated Hospital of Guangzhou University of Chinese Medicine.We have collected 8 spleen and stomach qi deficiency syndrome patients with myasthenia gravis,and 8 volunteers in school.Subjects were asked about the general information of subject and taken 4 mL Fasting peripheral venous blood.The mRNA expression of PGC-1 was detected by polymerase chain reaction PCR method.After ststistic processing,compared with group difference.3.Animal experimental studyFemale Lewis rats were randomly divided into 2 groups:Normal control group and EAMG.The EAMG rats were anesthetized and immunized subcutaneously with R97-116 Peptides for 6 weeks.The normal control group rats received the same emulsion except PBS was use instead of the peptide.At the end of the experiment 54 rats were left in the model group.The number of successfully established experimental model rats is 43 with EMG detection.The successfully established EAMG rats were randomly divided into model group,QJ decoction high dose group,QJ decoction middle dose group,QJ decoction low dose group by using a random number table method.Each group contained 8 rats.Two weeks after EAMG was confirmed,Chinese medical group was fed with Chinese herbs,while model control group and blank control group were given normal saline.The treatment group were given Qiangjijianli decoction respective in the dose of 23.4 mg/kg,15.6mg/kg BW· d and 7.8mg/kg BW · d.The amount of intragastric administration is 10ml/Kg BW,which was given once per day for 4 weeks.At the end of the experiment,observed general conditions,weight and the changes of PGC-1,NRF-1 and Tfam mRNA expression in skeletal muscle of EAMG rats.Results1.Clinical study1.1 This study included 159 patients,the average age of onset was 40.39±15.48 years old,male:femalewas 1:1.65,female patients accounted for 62.26%,moderate systemic 49.06%.1.2 In patients enrolled,67 cases of pale red tongue,pale and enlarged in 16 cases,accounted for 52.2%,58 white tongue moss cases,35 white greasy tongue moss cases,29 cases of thin and whitish tongue moss,accounted for 76.73%.Among all cases,thready pulse is the most common,a total of 111 patients accounted for 69.81%.Deficiency of spleen qi and stomach qiwith the highest proportion of 52.72%,spleen and kidney deficiencyaccounted for 23.27%,spleen deficiency and dampness heatsyndrome accounted for 22.01%.1.3 In 159 patients,the number of whom with thymoma is 76,accounting for 47.8%.The deficiency of spleen and kidney with thymoma in 24 cases,accounting for 64.86%.78 cases of patients with type Ⅱb,a total of 54 cases with thymoma which accounted for 69.23%.1.4 The PGC-1α mRNA expression in peripheral blood of MG patients with syndrome of deficiency of spleen qi and stomach qi group was significantly lower than that in normal control group(P<0.05)。2.Animal experimental study2.1 After the injection,fur of the model rats was not shiny and there was a significant difference between normal group and model group in body weight(P<0.05).Chinese medicine could improve clor and luseter of rats’ fur and caused the treatment group body weight gain,but it still lower than normal group,(P<0.05),There was a significant difference in body weight between the low dose group and the middle dose group((P<0.05),no significant difference between the high dose group and the middle dose group(P>0.05).2.2 Electron microscope observation of skeletal muscle,mitochondria and neuromuscular junction:The control group rats’ muscle fiber Z lines were arranged in order,the number of mitochondria was normal,the structure was complete,and the synaptic folds in the NMJ and muscle were not obviously abnormal.In the model group,there were sparse muscle fibers,muscle membrane hacklily shrank with decreased mitochondria,and sparse,diffluent,or absent NMJ folds.After interventionwith Qiangji jianli Decoction,the above pathology changes wre improved:muscle fiber Z lines were clear,the number of mitochondria increased and the structure was complete.The NMJ structure was close to normal.2.3 The mRNA expression of PGC-1α in skeletal muscle of EAMG rats was significantly lowerer than that in normal rats.(P<0.05).There was no significant difference among three treatment groups(P>0.05).2.4 The mRNA expression of NRF-1、Tfam PGC-1α in skeletal muscle of treatment groups rats were increased,but there was no significant difference between treatment groups and model group(P>0.05).Similarly,there was no significant difference among three treatment groups(P>0.05).Conclusion1.Clinical study1.1 Female patients were more common and moderate systemic type is the most common type.Pale red tongue,white tongue moss,thready pulse are the most common.The disease mainly could classified into three syndromes,syndrome of deficiency of spleen qi and stomach qi,syndrome of deficiency of spleen and kidney,syndrome of dampness-heat due to spleen deficiency.The main pathogenesis of the disease is deficiency of spleen and stomach.1.2 In patients with thymoma,spleen and kidney deficiency syndrome is the most common,the proportion of patients with thymoma in syndrome of deficiency of spleen qi and Stomach Qi was significantly lower than that of the other two syndromes,thymoma ratio in the type II B patients is the highest.The reaction of thymoma associated with disease severity.1.3 The abnormal expression of PGC-1 mRNA in peripheral blood may play an important role in the pathogenesis of MG.2.Animal experimental study2.1 EAMG rat models could be established via subcutanceous injections of man-made synthetical peptide of acetylcholine receptor α subunit 97-116.2.2 Qiangji jianli decoction could improve the general conditions of EAMG rats,including body weight,symtoms,clor and luster of their fur.2.3 There wre sparse mussy muscle fibre,decreasing mitochondria in EAMG rats.Qiangjijianli decotion could improve the pathology changes in muscle mitochondria and NMJ of the EAMG rats.2.4 Ther were a significantly reduceon the mRNA expression of PGC-1 alpha,NRF-1,Tfam,the three genes plays an important role in the pathogenesis of myasthenia gravis.Qiangji jianli decoction can obviously improve the symptoms of MG,and it is mainly through improving the expression of them. |
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