| Purpose:This experiment set early mature animal model of wetresistance in middle-jiao for the study of platform, designed to study wet resistance distribution and content of AQPs in animal model of gastrointestinal, exploring related signal transduction factor of wet resistance in middle-jiao and Pingwei Powder intervention development relations.to reveal related signals transduction factor’s mechanism of action, to open up a new way for Pingwei Powder regulation in metabolic disorder of water,which makes the systematic, deep, comprehensive understanding of Pingwei Powder on the research of mechanism effect. Methods:Using Huang Xiushen’s comprehensive model method formodeling, simulating a few general wet factors,such as "excessive outside wet blocking the spleen and stomach leading to food out of control,wet coming from the inner and emotional obstruction so as to gas is blocked and water does not carry", establish scientific and reliable wet resistance in middle-jiao animal model. Using immunohistochemical techniques for the determination of gastrointestinal tissue distribution of AQP, using enzyme-linked immunosorbent assay (ELISA) determination of gastrointestinal tissue, mucosal and serum AQP, AVP, VIP content and related signal transduction factor content. Result:1. gastrointestinal aquaporin expression and distribution of pathologicalcharacteristic of wet resistance in middle-jiao animal model.1.1The syndrome of wet resistance in middle-jiao distribution of expression ofAQPs increasing Compared with the blank group, after setting model AQP1in cardiac and intestinal mucosal layer increase the distribution of the expression; AQP2in muscular layer, outer membrane layer and the mucous layer of the large intestine end increase the distribution of the expression;AQP3in mucous layer of cardia, the mucous layer and outer membrane layer of the small intestine middle layer increase the distribution of the expression; the expression and distribution of AQP4in the gastrointestinal tract have no difference; AQP5at the end of the mucosal layer of the large intestine increase the distribution of the expression;AQP7in gastric muscular layer and outer membrane layer, in mucosal layer of the small intestine middle layer and at the end of the the large intestine increase the distribution of the expression;AQP9in outer membrane layer of the cardia increases increase the distribution of the expression.1.2The syndrome of wet resistance in middle-jiao distribution of expression of AQPs reductionCompared with the blank group, after setting model the expression and distribution of AQP1in the gastrointestinal tract have no obviously decreasing trend; the expression and distribution of AQP2in the middle membrane layer of the gastric body intermediate mucosal have a decreasing trend; the expression and distribution of AQP3in the gastrointestinal tract have no obviously decreasing trend; the expression and distribution of AQP4in the gastrointestinal tract have no obviously decreasing trend; the expression and distribution of AQP5in the outer membrane layer of the the middle of gastric body have a decreasing trend; the expression and distribution of AQP7in the outer membrane layer of the middle layer of the gastric body and small intestine have a decreasing trend; the expression and distribution of AQP9in the mucosal layer of gastric and the outer membrane layer of the middle of the stomach body and small intestine have a decreasing trend。 2.The Pingwei Powder’s intervention expression and distribution of the gastrointestinal aquaporin of wet resistance in the middle-jiao2.1enhancement effectCompared with the model group, after the Pingwei Powder treatment, the expression distribution of AQP1in the cardiac tissue under the mucous, the middle mucosa layer of gastric body and the middle mucosa layer of intestine enhance; the expression distribution of AQP2in the the middle mucosa layer of gastric body and the middle mucosa layer of large intestine enhance; the expression distribution of AQP3in the the middle of small intestine enhance; the expression distribution of AQP5at the end of large intestine enhance; the expression distribution of AQP9in the cardiac mucous and the middle of gastric body enhance.2.2inhibition effectCompared with the model group, after the Pingwei Powder treatment, inhibition AQP expression distribution of wet resistance in middle-jiao: inhibition AQP1expression distribution in the cardia mucosa; inhibition AQP2expression distribution in the midddle of lager intestine tissue under the mucous; inhibition AQP3expression distribution in the mucosa layer of cardia and the middle of lager intestine; inhibition AQP4expression distribution in the cardia mucosa.3. Determination of related regulation mechanism3.1Determination of long-time regulation mechanisms3.1.1The AQP content determination of wet resistance in the middle-jiao3.1.1.1The AQP levels of wet resistance in the middle-jiao RiseCompared with the blank group, after setting model, AQPO content in the middle of large intestine tissue increased (P>0.05); AQP2content in the large intestine end tissue increased (P<0.05); AQP3content in the middle of the small intestine tissue increased (P>0.05); AQP3content in the middle mucosal of large intestine increased (P<0.05).3.1.1.2The AQP levels of wet resistance in the middle-jiao decreasedCompared with the blank group, after setting model, AQPO content in the colonic tissue decreased (P<0.05); AQP1content in cardiac tissue decreased (P<0.01); AQP1content in the intermediate mucosal of stomach body decreased (P<0.05); AQP2content in the middle tissue of large intestine decreased (P<0.05); AQP3content in the middle of the small intestine tissue decreased (P<0.05); AQP3in the middle tissue of large intestine decreased (P<0.05):AQP9content in intermediate mucosal of the stomach body decreased (P<0.01); AQP9content in intermediate tissue of stomach body decreased (P<0.05).3.1.2The Pingwei Powder’s intervention expression and distribution of the gastrointestinal aquaporin content of wet resistance in the middle-jiao3.1.2.1enhancement effectCompared with the model group, after the Pingwei Powder treatment, the AQPO content in the middle of intestinal tissue increased (P>0.05); AQP1content in the intermediate mucosal of stomach body increased (P<0.01); AQP2content at the end of large intestine tissue increased (P>0.05); AQP3content in the middle of the small of intestinal tissue increased (P>0.05); AQP3content in the colon tissue increased (P>0.05); AQP3content in rectal tissue increased (P<0.05); AQP9content in the stomach body intermediate mucosal increased (P<0.01); AQP9content in the stomach body intermediate tissue increased (P>0.05).3.1.2.1inhibition effectCompared with the model group, after the Pingwei Powder treatment, AQP2content in the middle of large intestine tissue decreased (P>0.05); AQP3content in large intestine intermediate mucosa decreased (P>0.05).4. Determination of short-term adjustment mechanism4.1The related hormone of wet resistance in the middle-jiao and signal transduction pathway of AC-cAMP-PKA in relationships (stomach)Hormone regulation mediated by AVP and VIP, the various factors role in cell signal pathways such as:AC, cAMP, PKA content changes are demonstrated in the same directionwith AVP, VIP content change trend.4.2The related hormone of wet resistance in the middle-jiao and signal transduction pathway of PLC-IP3/DG-CaM/PKC signal transduction in relationships (intestine)After setting the model, both AVP and VIP content in serum are decreased, and a variety of signal molecules which effect on signal transduction pathways such as:PLC, IP3, CaM, Ca2+. Compared with AVP and VIP content, after setting the model, AVP and VIP content decreased.the content of PLC, IP3, CaM, Ca2+increased, but the natural recovery group declined, so the signal transduction pathway of intestinal and gastric are consistent (possibly the physiology function of the gastric and large intestine are different). [Conclusion] Setting model of accumulation of dampness in middle-jiao model and after drug delivery, not only affect the distribution changes of gastrointestinal aquaporin, but also caused the content change of gastrointestinal aquaporin. The molding methods and through the effect of Pingwei Powder on aquaporin content and distribution changes to cause the rats in water electrolyte change. Around the aquaporin hormones such as AVP, VIP and related signal transduction pathways in a variety of signaling molecules also produced corresponding change. Descript that Pingwei Powder’s effects on rats in fluid and electrolyte through the action of the aquaporins as well as these hormones and signaling molecule mechanisms to achieve. |
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