The Relevant Research Between Energy Metabolism Dysfunction And Mitochondrial Membrane Potential And Atpase On Spleenasthenic Syndrome

Background and ObjectiveIn Traditional Chinese Medicine,the spleen masterring the movement and transformation,the spleen govermenting blood,and the spleen masterring muscle and the arms and legs,spleen provides the materiel basis of the acquired constitution,is the source of growth and development.In morden medicine,mitochondria,which almost existing in the whole human body tissue and cell,provides energy for cell biological oxidation of every system of body,called"the production base of the ATP" and "the power factory of the cell".The spleen is in common with mitochondria in many aspects,such as function.The splenasthenic syndrome is the most common type in TCM clinical.This study in order to temper deficiency syndrome in patients with myasthenia gravis as the research object,to observe the temper deficiency syndrome myasthenia gravis patients peripheral blood lymphocyte mitochondrial membrane potential changes;To spleen-qi deficiency model rats as the research object,which were treated by Sijunzi decoction,then to observe the mitochondrial membrane potential of the hepar and spleen tissue and ATPase activity of skeletal muscle tissue.By adopting the combination of clinical and experimental research methods,from the function of mitochondria changed to explore mechanism of spleen deficiency energy metabolic disorders and effects of the Spleen yi-qi to the decoction,provide guidance for clinical drug use and improve the curative effect.MethodsTo collect 2.5ml peripheral blood of the spleen-qi deficiency patients with MG,who were admitted to spleen and stomach department of the first affiliated hospital of Guangdong University of TCM from July 2013 to March 2014(as trial group),10 cases for each group,Isolating the lymphocyte from peripheral blood and detect the mitochondrial membrane potential of it by using flow cytometry.To choose 42 SPF SD rats,divided into two groups according to random number table method,only 10 model group 32 only.The model group were given abdominal subcutaneous injection with 0.5mg · kg-1 · d-1 reserpine to make spleen-qi deficiency syndrome model rats,normal group rats subcutaneously amount of normal saline,a total of 8d.After the building,observing rats weight and so on general situation and the determination of urinary D-xylose excretion rate,assessment of building success in rats.According to random number table method,the selection of normal rats 7,14 model group rats,and randomly divided into group,spleen-qi deficiency group and Si junzi decoction group in the two groups,each group of 7.Si junzi decoction group with Sijunzi decoction gavage,at the dosage of 10.5g/Kg/d,model group and normal group to the irrigation amount of normal saline,filling and 3 weeks after the death of rat.Prepare single-cell suspension of tissue of spleen and liver,using flow cytometry instrument detection in the rat liver,spleen organization level of mitochondrial membrane potential.Isolating the mitochondria from skeletal muscle tissue of the rats,then to detect the mitochondrial Na+-K+-ATPase?Ca2+-Mg2+-ATPase activity of the tissues by lipase-catalyzed method.Results1.Spleen deficiency syndrome myasthenia gravis patients peripheral blood lymphocyte mitochondrial membrane potential was significantly lower than normal control group(P<0.05).2.The rats of the spleen-qi deficiency group showed obvious typical spleen-qi deficiency syndrome,such as weight reduction,emaciation,anorexia,rectocele,lethargy with narrowed eyes,hogback,huddling up their bodies,spiritual burnout,et al,but without remarkable change of anal temperature.Determination of spleen-qi deficiency group,D-xylose level significantly lower than normal group(P<0.05).Symptoms of spleen-qi deficiency after applying sijunzi decoction can obviously improve,urine D-xylose level also increased significantly(P<0.05).3.Comparison on the mitochondrial membrane potential of the rat-tissues of liver and spleen:1)The mitochondrial membrane potential level of liver tissue of spleen-qi deficiency group is obviously lower than that of normal group(P<0.01);the level of Sijunzi decoction group is significantly increased than spleen-qi deficiency group(P<0.05),but still lower than normal group(P<0.05).2)The mitochondrial membrane potential level of spleen tissue of spleen-qi deficiency group is obviously lower than that of normal group(P<0.01),the level of Sijunzi decoction group is significantly increased than spleen-qi deficiency group(P<0.05),but still lower than normal group(P<0.01).4.Comparison on the activity of mitochondrial Na+-K+-ATPase?Ca2+-Mg2+-ATPase of the rat skeletal muscle tissue:spleen-qi deficiency group is obviously lower than normal group(P<0.01 or p<0.05),Sijunzi decoction group is significantly higher than spleen-qi deficiency group(P<0.05),but it is gently lower than normal group.ConclusionMitochondrial membrane potential level of peripheral blood lymphocyte of the spleen-qi deficiency MG patients was lowerer than normal control group,showed that mitochondrial membrane permeability increased,ATP synthesis,cell energy metabolism disorder,therefore,spleen-qi deficiency syndrome is closely related to the mitochondrial membrane potential.Compared with normal group,model of rat liver,spleen-qi deficiency organization mitochondrial membrane potential level significantly decreased,after treatment with sijunzi decoction can obviously increase the liver,spleen tissue mitochondrial membrane potential,improve mitochondrial function.Spleen-qi deficiency model in rats skeletal muscle mitochondria Na+-K+-ATPase?Ca2+-Mg2+-ATPase activity significantly lower than the normal group,after treatment with sijunzi decoction invigorating spleen yiqi can obviously improve the activity of ATP enzyme,improvement of energy metabolism.