| BackgroundAccording to the theory of Chinese traditional medicine (TCM), spleen is a functional structure unit which includes some organs’physical functions, such as stomach, pancreas, liver, spleen, and even the brain. Thus, it is a comprehensive functional unit with multiple organs and systems. So, spleen asthenia syndrome involves the physiological functioning disorders in the systems of digestion, immune, endocrine, neurology, sports. Research shows that the Qi-activating and Spleen-strengthening herbs (ep. Panax ginseng C. A. Mey, Radix Codonopsis, Astragalus membranaceus Bunge, Hedysari, Sijunzi decoction, Liujunzi decoction and Buzhong Yiqi decoction) have good effect to prevent and treat spleen asthenia syndrome.In the theory of living systems and signal transduction, biological phenomena (physiological and pathological state) is actually the consequence of mutual signaling and interaction between the inside and outside of the body, among different systems, tissues and cells. This coincides with the connotation of Holism theory and five-organs holism theory of TCM. The key molecules in Ca2+/CaM signal pathways have many biological functions which closely associated with such physiological functions as regulating cellular enzymes activity, muscle contraction and relaxation, regulating nerve system function, stimulating secretion, regulating sugar metabolism, regulating prostaglandin and synthesis and release of insulin, increasing brain long-term memory, etc. Recently, academic researches into the essence of spleen asthenia syndrome tend to study from multi-systems, multi-viscera, multi-angle, multi-level (cellular, molecular and gene level).Analyzed from syndrome of TCM, elements of spleen asthenia syndrome include: pale white or yellow complexion, poor appetite and dyspepsia, abdominal distension or diarrhea, severe abdominal distension after meal, fatigue, less words, tired limb and body, even thin body, muscle atrophy, less movement, susceptible to cold and wind, weak and sick physique, obesity with edema, whitish and pale tongue with white fur, thin or weak pulse. The spleen asthenia syndrome involves the disorder or physiological hypofunction in digestion, immune, endocrine, neurological, sports and other systems, so the signs and symptoms tend to be generalized in the body. Modern research proves that Ca2+/CaM-CaMK II plays an important role in many diseases and dysfunction of multiple organs. What’s more, it can be conjectured from the studies relative to physiological functions and spleen asthenia syndrome which is based on Ca2+/CaM signal pathways that there exists some relations between the physiological functions of Ca2+/CaM signal pathways and TCM theory of "Spleen governs digestion and absorption, rule blood, nourish muscles and limbs, and thought in spiritual activities". Still, the subject needs to be researched further.ObjectiveOn the basis of TCM concept of entirety, and clinical syndrome factors and pathogenesis evolution law, the syndrome patterns of "soma generalization effect" were put forward in the thesis. And according to the theoretical knowledge about Ca2+/CaM and its related signal pathways, a hypothesis that Ca2+/CaM and its related signal pathways probably are effective on the soma generalization effect of spleen asthenia syndrome was established, in order to study the pathological connotation of spleen asthenia syndrome, syndrome foundation, and the interposition of the intervention effect of Sijunzi decoction, extract from hedysarum polybotrys on the signal pathways.MethodsBy adopting the methods of consuming energy with bitter cold, swimming exhaustion and irregular feeding to establish spleen asthenia syndrome model with rats, then treat them with extract from hedysarum polybotrys and Sijunzi decoction. And by detecting their general existence, daily average food consumption, daily average gaining in weight, swimming endurance time, gastric remnant rate, intestinal propulsion rates, the content of serum D-xylose, gastrointestinal hormone GAS, MOT, SS and VIP in small intestine tissue, the rat models were evaluated. Based on the successful modeling, after applying Qi-activating and Spleen-strengthening herbs, such as Sijunzi decoction and extracts from hedysarum polybotrys to the rats, a detection on fasting blood-glucose, serum insulin level, and enzyme activity related to metabolism in different tissue was carried out. Also, confocal laser technology was used to test cellular [Ca2+]i concentrations in small intestine, pancreas, skeletal muscle and liver tissue. Real-time PCR and Western blotting technique were used to test CaM, CaMK Ⅱ from Ca2+/CaM signal pathway and related protein expression in small intestine, pancreas, skeletal muscle and liver tissue of spleen asthenia rats.Results1.The rats in the spleen deficiency group gradually showed spleen asthenia symptoms such as laziness and poor appetite, sparse hair, thin with arch back, slow to move, decrease in daily food intake, slow increase in body weight, shit more frequently, filthy peri-anus, even prolapse. Swimming endurance time obviously decreased (P<0.05), gastric remnant rate increased and small intestinal propulsion rates decreased (P<0.05), and spleen deficiency21d group changed significantly (P<0.05).2.In spleen deficiency7d model group, FBG and serum insulin tended to lower slightly, but there were no statistical difference (P>0.05). In spleen deficiency14d and21d model group, FBG and serum insulin tended to decrease significantly, especially in the spleen deficiency21d model group. As the molding time prolongs, the activity of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and SDH in small intestine, pancreas, skeletal muscle and liver tissue of the spleen deficiency rats decreased significantly (P<0.05), but the activity of LDH increased significantly (P<0.05).3.[Ca2+]i concentration in the cells of small intestine increased significantly (P<0.05), but decreased in pancreas, skeletal muscle and liver tissue (P<0.05). The expression of CaM, CaMK Ⅱ and CaM, CaMK Ⅱ, p-CaMK Ⅱ in small intestine tissue increased (P<0.05), but decreased in pancreas, skeletal muscle and liver tissue (P<0.05).4.The extract from hedysarum polybotrys and Sijunzi decoction improved general existence of rats, increased swimming endurance time and content of serum D xylose, decreased gastric remnant rate, slowed down the small intestinal propulsion rates, and the content of small intestine tissue, serum gastrointestinal hormone:increased the GAS, MOT levels but decreased the SS, VIP levels, and Sijunzi decoction was more significant (P<0.05).5.The extract from hedysarum polybotrys and Sijunzi decoction significantly increased FBG and serum insulin levels of the spleen deficiency rats (P<0.05), also improved the activity of the Na+-K+-ATPase, Ca2+-Mg2+-ATPase and SDH (P<0.05), decreased the activity of LDH (P<0.05) in small intestine, pancreas, skeletal muscle and liver tissue, and Sijunzi decoction was more significant.6.Sijunzi decoction and Extract from hedysarum polybotrys significantly decreased [Ca+]i in small intestine, increased [Ca2+]i in pancreas, skeletal muscle and liver. Sijunzi decoction and Extract from hedysarum polybotrys decreased the expression of CaMmRNA, CaMKⅡmRNA and CaM, CaMKⅡ, p-CaMKⅡ in small intestine (P<0.05), increased the expression in pancreas, skeletal muscle and liver tissue (P<0.05), and the Sijunzi decoction had more significant effect.Conclusion1. The methods of consuming energy with bitter cold, swimming exhaustion and irregular feeding can build the ideal spleen deficiency animal model. The spleen deficiency rats showed a poor living condition compared with the normal rats. With the time extension of building, the model rats showed decreased food intake, slowly daily weight increase, decreased anal temperature and swimming endurance time. Gastric remnant rate and intestinal movement increased, but the level of the serum D xylose, FBG, serum insulin decreased, caused secretion disorder of gastrointestinal hormone GAS, MOT, SS and VIP.2.The FBG and serum insulin level of spleen deficiency rats decreased, with lower level of glucose metabolism. And, the activity of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and SDH decreased obviously in small intestine, pancreas, skeletal muscle and liver tissue, while the activity of LDH increased obviously which showed energy metabolism disorder in small intestine, pancreas, skeletal muscle tissue and liver tissue.3.There is a difference in the expression of Ca2+/CaM-CaMK Ⅱ signal pathway during the course of the spleen deficiency, mostly higher expression in the small intestine,[Ca2+]i concentration increased, the expression of CaM, CaMK Ⅱ genes and protein increased, mostly low expression in the pancreas, skeletal muscle tissue and liver tissue,[Ca2+]i concentration decreased, also, the expression of CaM, CaMK Ⅱ genes and protein decreased.4.The generalization effect in the spleen deficiency body involves the disorder of the body’s glucose metabolism balance, also the disorder of energy metabolism in small intestine, pancreas, skeletal muscle and liver. And, there are different expression of the Ca2+/CaM-CaMK II signal pathway in different tissue of the spleen deficiency rats.5.The extract from hedysarum polybotrys and Sijunzi decoction can improve general existence of rats, increase swimming endurance time, adjust the secretion of gastrointestinal hormone GAS, MOT, SS, VIP and the function of the gastrointestinal transition; increase the level of the spleen deficiency rats’plasma glucose of limosis, serum insulin, and the related enzyme activity which is metabolized in small intestine, pancreas, skeletal muscle and liver tissue; regulate the normal expression of CaM, CaMK II genes and CaM, CaMK II, p-CaMK II protein. Sijunzi decoction had more significant effect. As a result, it showed that, under the guidance of Chinese medicine theory, the herbal decoction had advantages over single herb extract. |
PDF Full Text Request