| Objective: It is well known that predominance and quantization of splenogastric hygropyrexia syndroma are the keys of diagnoses and treatment. In this study,animal models of splenogastric hygropyrexia syndroma with predominance of wet or heat are made by exerting factors such as wet heat environment, high fat and glucose diet, alcohol, decoction of monkshood and rhizoma Zingiberis and pathogenic microorganism, testified by the effectiveness of baihu and Rhizoma Zingiberis Atracylodis decoction and QingXiangSan,and quantized by observation of the changes of symptom, physical sign and indexes of biochemistry, immunity and pathology of the animal so as to elucidate the mechanism and pathological foundation of splenogastric hygropyrexia syndroma.,furtherly to reveal the essence of splenogastric hygropyrexia syndroma and establish the therapeutic foundation on serial diseases including in hygropyrexia syndrom.Methods: To study the animal models of splenogastric hygropyrexia syndroma with predominance of wet or heat by observing the effect of few adding factors , therapeutic effect of herb decoction on five groups of SD mouse just like the group of control, predominance of wetness , predominance of heat, baihu and Rhizoma Atracylodis decoction and.eight animals each group.To quantizesplenogastric hygropyrexia syndroma with predominance of wetness or heat through the methods of biochemistry > immunity and electron microscope to detect AQP2 in urine and expression in kidney> HSP70 in lymphocyte, motilin and gastrin in serum, Ultrastructural change of spleen and indexes of immune histochemistry suchasAI> Bcl-2andP53. Results and Analysis l.The making of animal model and experimental observation(l)The making of animal modelThe group of contrast(group A): common mixed feed and normal environment of 20-28 "C temperature, 50-70% relative humidity.The group of predominance of wet(group B): high fat and glucose diet which includes 12% grease > 8% honey for 10 days ,next wet heat environment of 33+2°C temperature, 90-100% relative humidity in a cabinet,simultaneously alcohol diet lavaged (2ml /200g> once a day) for eight days,next 2ml/200g colibacillus lavaged and lml/200g 24h later.next normal environment like the control.,next distilled water lavaged (8ml/200g , twice a day) 12h later for 7 days.then the animal were killed.The group of predominance of wet(group C):the method is just like group B,except that monkshood and Rhizoma Zingiberi (2ml /200g, once a day) is in place of the alcohol.The group of QingXiangSan(group D): the method is just like group B,except that QingXiangSan iseed in place of distilled water.The group of baihu and Rhizoma Atracylodis decoction (group E): the method is just like group C,except that baihu and Rhizoma Atracylodis decoction is in place of distilled water.(2) Experimental observationTo observe mouses mind state % activity > numbers and properties of fecal > body temperature and the quantity of dietThe animals of group A have no changesregarding mind state -. activity > numbers and properties of fecal.these of group B and C appear less quantity of diet, loose stool -. dirty crissum from 5 days later .but no different body temperatures, afer they were moved into the wet-heat cabinet and lavaged with alcohol or decoction of monkshood and Rhizoma Zingiberi these of group B appear loose fecal -> dirty crissum, lassitudes idtex dullness > less quantity of diet sequentially lightly High rectal temperature., These of group C have dry stool with mucosa, pugnacity, less quantity of diet,sequentially > lightly High rectal temperatura.After lavaged with colibacillus,the syndroma furtherly aggravates,but after the corresponding drugs given and the syndromaes of these of group D and E significantly ameliorate.Regarding to the mind state > activity x properties of fecal n body temperature of each group on models of splenogastric hygropyrexia syndroma with predominance of wetness and heat, we can know that the animal models of splenogastric hygropyrexia syndroma with predominance of wetness and heat are objective.2.The change of motilin and gastrin in serumThe change of motilin and gastrin can reflect status of digestive function objectively.and the detection of these can reveal the digestive function change of splenogastric hygropyrexia syndroma with predominance of wet or heat.As to the motilin,these of group B increase and group C decrease.there are significant difference in comparison with group A, D and E respectively (P<0.05) ,these of group A have no significant difference in comparison with group D and E (P>0.05) .as to gastrin,these of group B and group C decrease,and group C less,but there is no significant difference between these of group B and C (P>0.05) ,and significant difference in comparison with group Ax D and E (P<0.05) , these of group D and group E have no significant difference with group A (P>0.05) .we can come to a conclusion that the drugs applied in group D and E can effectively modulate thedensity of gastric stimulate gastric acid secretion promote Gastrointestinal mucosa growth and make the density of gastrin stay the normal status.3.The change of AQP2 in urine and expression in kidneyAQP2 can regulate the metabolism of body fluid. It is important for the formation of splenogastric hygropyrexia syndroma with predominance of wet and the quantization of wet and heat,especially the quantization of wetas to the AQP2 in urine .these of group C decrease,and group D increase.there are significant diffirence in comparison with these of group As E and D (P<0.05) .As to the expression in kidney,these of group B are less than these of group A> group E and group D,and these of group C is the most.we come to a conclusion that baihu and Rhizoma Atracylodis decoction and QingXiangSan can regulate AQP2 so as to regulate the metabolism of body fluid,and this maybe the mechanism of dehygrosis.and the important effect of AQP2 on the formation and quantization of splenogastric hygropyrexia syndroma with predominance of wet testify the objectiveness of the animal model of splenogastric hygropyrexia syndroma with predominance of wet or heat which we have made.4.Expression of HSP70 in lymphocyteHeat stress proteins is induced to synthetize when organism is under heat stress or other factors that can injury the cells so as to improve the capability of thermoresistance and resistance.some studies have proved that there is relationship between the splenogastric hygropyrexia syndroma with predominance of heat and HSP70.In our study the expression of HSP70 in lymphocyte of group B and group C increase more than these of group A (P<0.01) ,and these of group C are significantly more than these of group B (P<0.05) .We come to a conclusion that the change of HSP70 under the factors such as the wet heat environment high fat and glucose diet> alcohol -. decoction of monkshood and rhizoma Zingjberis and pathogenicmicroorganism .especially the colibacillus, can be a important index to quantize the splenogastric hygropyrexia syndroma with predominance of heat or wet.and baihu and Rhizoma Atracylodis decoction and QingXiangSan can regulate the expression of HSP70 and this maybe the mechanism of heat-clearing effect. And this is also testifies the feasibility of the animal model.5. infrastructure observation of spleenWith electron microscope, we get the message of Ultrastrucrural change of spleen of splenogastric hygropyrexia syndroma with predominance of heat or wetand these of group B and group C have significant change.the main appearance is just like these that narrow splenic sinusoid, endotheliocyte swelling, the damaged structure of macrophage, lymphocyte, plasmacytic, cell organs swelling or atrophia like mitochondria, lysosome, rough endoplasmic reticulum , karyopyknosis etc.Spleen is a important immune response organ.these of group B and group C have significant change ,and this is the pathomorphology foundation of immune function disorder and low resistance against macroorganism,these of group D and E have little changes ,thus we can conclude that baihu and Rhizoma Atracylodis decoction and QingXiangSan can regulate immune function.6.Observation of index (AI, Bcl-2 , P53 )of immune histochemistryWe investigate the pathological change of gastric epithelium mucosae of splenogastric hygropyrexia syndroma with predominance of heat or wet through the detection of apoptotic index AI by method of TUNRL and cell apoptosis associated protein such as Bcl-2, P53 by immune histochemistry method.AI in group B and C are significant higher than these of group D, E and A (P<0.05) .but there are no significant differene between these of group D, EandA (P>0.05) .Conclusion:1.Animal models of splenogastric hygropyrexia syndroma with predominance ofwet or heat were made by exerting factors such as wet heat environments high fat and glucose diets alcohoK decoction of monkshood and rhizoma Zingiberis and pathogenic microorganism and referring to the indexes of biochemistry > immunity and pathology and molecular biology.2. The change of motilin and gastrin can objectively reflect the status of digestive function.The gastrin of group B increase and group C decrease;The motilin of group B and C decrease.and these of the group C is more,all these reflect the disorder of splenogastric hygropyrexia syndroma.3. AQP2 can regulate the metabolism of body fluid,and in our study .these in urine of group B decrease and group C increase.The experiment of these in kidney of group B decrease more than these of group A and E,wherever these of group C increase,therefore it can be one of the most important indexes to quantize splenogastric hygropyrexia syndroma with predominance of wet or heat.4.The increase of HSP70 can reflect the degrees of inflammation.and the increase of HSP70 of the animal model and the more increase of these of splenogastric hygropyrexia syndroma with predominance of heat than that of wet demonstrate that it can quantize splenogastric hygropyrexia syndroma with predominance of heat.5.The investigation of Ultrastructural change of spleen with electron microscope reflects the animal model's spleen pathological change,and splenogastric hygropyrexia syndroma with predominance of heat or wet is different from each other.6. The apoptosis of gastric epithelium mucosae of splenogastric hygropyrexia syndroma significantly strengthened the apoptosis regulatory related gene like P53, Bcl-2 expression also strengthen,we presume that the expression of P53 strengthen higher than Bcl-2.7.The effect of baihu and Rhizoma Atracylodis decoction and QingXiangSan on the corresponding animal model suggest that they can effectively improve the symptom and physical sign >. regulate the AQP2 function in urine and kidney and the expression of HSP70> maintain the integrity of cell structure and function regulate serum motilin and gastrnu improve immune function and recovery capability ^ regulate the apoptosis of gastric epithelium mucosae and protect the gastric mucosae.The result of study shows : The utterly effect on many factor such as the circumstances of wetness and hot , fat sweet dieK medicine and pathogenic microorganism , set up the animal models of splenogastric hygropyrexia syndroma with predominance of wetness and heat, after exerting white spirit and monkshood and doing the ginger decoction separately, cause wet to lay particular stress on spleen and stomach damp and hot card old mouse model and heat lay particular stress on spleen and stomach damp and hot card old mouse model, solicit through observing clinical symptom and body, two above-mentioned models accord with clinically; Use classics , experience side (fragrant to break up , add pines and cypresses of verdant green soup by white tiger ) disproof above-mentioned two objectivity of model also through serial measuring of index. The spleen is wet in the damp and hot card of spleen and stomach and theory taking spleen and stomach as the centre of stomach energy, describe a lot in classics, but lack the index of objectifying, we carry on the discussion from the stage construction in the experiment for this: Exceed the pathology change of the micro-structure from the spleen; The biochemical index plasma stomach is moved plainly , the stomach secretes plain change and serial immune groups like stomach mucous membrane AI, such essence of reflecting the damp and hot card card of spleen and stomach of consistency as the expressions of P53 , Bcl-2 ,etc. lie in the imbalance and disorder of spleen and stomach digestive function, it is theories of the centre to confirm the spleen and stomach. Spleen seriousquantization not wet as body it it appears that water is not normal in supersession in liquid,it correlated with AQP2 (the wet when not laying particular stress on AQP2 content reduce urine obviously, it is hot the urine AQP2 content increases while laying particular stress on), so can be as measuring one of the indexes of resolving dampness; Stomach energy serious between quantization and HSP70 is reflect laying particular stress on relevant hot to express. In surn,to make animal models of splenogastric hygropyrexia syndroma with predominance of wet or heat by exerting factors such as wet heat environment high fat and glucose dieU alcohol n decoction of monkshood and rhizoma Zingiberis and pathogenic microorganism,apply the theory of traditional Chinese medicine.intrgrate contemporary experimental method,so as to explore the animal model of splenogastric hygropyrexia syndroma with predominance of wet or heat from different respects just like biochemistry > immunity> pathology and molecular biology;and furtherly interpret the mechanism and pathological foundation of splenogastric hygropyrexia syndroma with predominance of wetness and heat. |
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