| Purpose:Spleen transportation is the most important physiological function in the spleen system.One of spleen transportation is to transform nutrients from food into essence and spread it to all organs and tissues of the whole body.As mentioned in “Su Wen”: Drink into the stomach,swim and overflow the essence Qi,lose to the spleen,spleen qi and essence,and return to the lung.The other transportation is to balance the metabolism of water and fluid with lung,kidney,triple energizer and bladder.It is also known as “Su Wen”: All wet and swollen are all spleen.Therefore,when the spleen fails to be healthy(spleen deficiency),the patient shows symptoms of systemic hypofunction,for example,weak,abdominal distention and diarrhea.Studies have shown that the dysfunction of organs caused by spleen deficiency syndrome is closely related to insufficient energy supply of organs.In the five viscera,the heart and spleen are closely related.In the five elements,the heart belongs to the fire,the spleen belongs to the earth,and fire can give birth to the earth,so the relationship between the heart and the spleen is mother and son.Heart-fire can warm the spleen,and the operation of the spleen depends on the warmth of the heart-yang.If the spleen and earth run continuously,the Qi and blood and the subtle substances can be changed to make the heart healthy.Abnormal functioning of the spleen and abnormal cardiac function are related to the poor supply of cardiac substances.Glucose is one of the three major energy substances in human life activities.It plays an important role in energy metabolism and body composition.Glucose is a polar molecule that requires glucose transporters(GLUTs)-mediated facilitated diffusion in order to enter the cell through the phospholipid bilayer structure of the cell membrane,and produce adenosine triphosphate(ATP).ATP is the most direct energy substance of the organism.Glucose transporter 4(GLUT4)is the most important glucose transporter in cardiomyocytes and responsible for glucose transport under stress.AMP-activated protein kinase,known as the energy receptor of the body,it is located upstream of GLUT4 and plays a positive regulatory role in GLUT4.Before that,the research group has shown that: glucose transporter 5(GLUT5)and glucose transporter 1(GLUT1)mRNA and protein expression in jejunum of spleen deficiency rats were significantly decreased,resulting in impaired glucose absorption,which is one of the reasons for insufficient energy supply.Another experiment in this research group proves that: Under the condition of spleen deficiency,the expression of glucose transporter1(GLUT1),glucose transporter 1(GLUT3)mRNA and protein expression in hypothalamus of rats were significantly decreased,resulting in insufficient energy supply of hypothalamus,affecting the expression of brain intestinal peptide(β-EP,CCK,VIP),resulting in abnormal gastrointestinal digestion,absorption,secretion,movement and other functions.The study on the expression level of GLUTs in different tissues of rats with spleen deficiency syndrome opens up a new way for the study the essence of spleen deficiency syndrome.Heart and spleen are closely related,and spleen deficiency can lead to cardiac function decline.Spleen qi deficiency is the foundation of other spleen deficiency syndrome,and is very important for the study of spleen deficiency syndrome.What is the relationship between the spleen qi deficiency,cardiac function and GLUT4 expression level in cardiac muscle tissue?This paper is guided by the theory of spleen transportation,starting from the spleen qi deficiency affect myocardial metabolism,glucose transporter as the starting point,investigate whether spleen qi deficiency through the AMPK pathway influent the expression of GLUT4,thereby inhibiting glucose transportion,resulting in heart muscle cell morphology change and abnormal heart function.Sijunzi pill is a classic medicine for invigorating spleen and replenishing qi.In addition,by giving the rats Sijunzi pill gavage treatment to observe the effects of Sijunzi pill on spleen qi deficiency rats above detection index,To observe the improvement of cardiac function and myocardial cell morphology in rats with spleen qi deficiency,and test GLUT4 and AMPK protein expression,investigate the expression levels changes of GLUT4 and AMPK in rats with spleen qi deficiency.Material and method:In this experiment,24 Male SD rats weighed(200 ± 10)g were selected,divided into 3 groups,and 8 rats in each group.They were control group,spleen qi deficiency model group and Sijunzi pill group.Exhausted dietetic and swimming method wasused to make spleen qi deficiency model,spleen qi deficiency syndrome model shows eat less,fatigue,weight loss,hair soft and lustrous,faeces soft.Sijunzi pill treatment group conversion according to body surface area,spleen qi deficiency treatment group rats were gavaged with0.648 g /100 g /d Sijunzi pill suspension,the control group and spleen qi deficiency model group rats were gavaged equal distilled water 14 days.By using the high-resolution ultrasound system of small animals,color Doppler echocardiography was performed in rats,and left ventricular ejection fraction and shortening fraction were measured in each group.In addition,the ultrastructural changes of myocardial tissue in each group were examined by transmission electron microscope.Then Western blot and Real-Time PCR were used to detect GLUT4 and p-AMPKa expression in the three groups rats.Results:1.The evaluation of modelAfter spleen qi deficiency model established,the model was evaluated by measuring the rats weight,anus temperature and food amount.The reseen qi defults showed that spliciency rats weight was 194.25 ± 4.76 g,rectal temperature was 36.52 ± 0.24 ℃,food intake was16.32 ± 0.70 g,the index was significantly lower than the control group rats.The weight,rectal temperature and food intake of Sijunzi pill treatment group rats were significantly improved,were 247.10 ± 4.55 g,37.04 ± 0.33 ℃,28.41 ± 1.00 g.2.Comparison of cardiac function in each group ratsThe heart function of the three groups rats showed that compared with the control group,left ventricular ejection fraction,fractional shortening of spleen qi deficiency group rats was significantly decreased,and left ventricular ejection fraction,fractional shortening of the Sijunzi pill group were increased significantly,but still lower than the control rats.3.The comparison of ultrastructure of myocardium in each group ratsIn this study,transmission electron microscopy was used to observe the spleen qi deficiency effect on rat myocardium ultrastructure.The results showed that the myocardial fibers in the control group were arranged regularly,well structured,and the intercalated plate structure existed between the myocytes.Compared with the control group,the myocardial fibers in the spleen qi deficiency group were regular,the muscle fibers arranged loosely(edema),the number was reduced,and some of the muscle fibers were dissolved.The number of myocardial fibers in Sijunzi pill group was regular and the number increased,and most of the intercalated plate could be seen.4.The changes of GLUT4 expression in each group rats’ myocardiumReal-Time PCR results showed that compared with the control group,GLUT4 mRNA expression in the heart tissue of spleen qi deficiency group rats was significantly reduced.And GLUT4 mRNA expression in the heart tissue of Sijunzi pill group was significantly higher than that in the spleen qi deficiency group.Western blot and statistical analysis showed that compared with the control group,the GLUT4 protein expression in the heart tissue of the spleen qi deficiency group was significantly reduced,while the GLUT4 in the heart tissue of Sijunzi pill group was significantly higher than that in the spleen qi deficiency group.5.The changes of p-AMPKa protein expression in each group rats’ myocardiumIn this study,Western blot was used to investigate the p-AMPKa expression changes in the each group rats’ heart tissue.The results showed that compared with the control group,the p-AMPKa protein expression in heart tissue of spleen qi deficiency rats was significantly reduced.Compared with the rats in spleen qi deficiency group,the protein expression of p-AMPKa in heart tissue of Sijunzi pill group was significantly increased.Conclusion:Under spleen qi deficiency condition,the cardiac function of rats reduced;the ultrastructure of rats’ myocardial tissue changed;GLUT4 expression in rat heart tissue reduced;p-AMPKa expression in rat myocardial tissue reduced.Sijunzi pill could increase GLUT4 and p-AMPKa expression in rat myocardial tissue with spleen qi deficiency;then improve the glucose transportion in the myocardium of rats with spleen qi deficiency;ultimately improve the cardiac function of rats. |
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