The Method Of Invigorating Spleen And Replenishing Qi Uses CAMP/PKA To Study The Regulation Mechanism Of Gastrointestinal Hormones In Rats With Spleen Deficiency And Spleen Failure

Purpose:Study on the TCM etiology and pathogenesis,including dietetic disturbance and excessive fatigue,this paper shows the spleen-qi deficiency syndrome model rats.It discusses the replicating and evaluating of spleen-qi deficiency syndrome model rats in view of behavior science,pathology and molecular biology,which is based on the anti-evidence groups of traditional Chinese medicine.The author researches on the biological metabolism of spleen-qi deficiency syndrome through cell damage,gastro-intestinal peptide hormones,energy metabolism and ion channels after testing MDA,GSH-Px,5-HT,VIP,respiratory chain complexes I and IV,Na⁺-K⁺ATPase,Ca²⁺-Mg²⁺ATPase,and cAMP?PKA of the rats.Followed by the theory of spleen image through the spleen-invigorating and qi-enhancing method,this paper studies on improving spleen deficiency and spontaneity of spleen-qi deficiency syndrome by replenishing Spleen-qi Deficiency based on the relieving energy metabolism abnormalities,anti-lipid peroxidation damage,and regulating gastrointestinal hormone secretion,which is at the core of cAMP?PKA signaling pathway.Materials and methods:Healthy SPF grades and SD rats are used in this study.the model is performed as composite factors with the method of fuzzy mathematics and the method of supplement of spleen and Qi.The changes of ileal mucosa in morphology are observed with HE staining.The ileal tissue respiratory chain complexes I and IV,Na⁺-K⁺ATPase,Ca²⁺-Mg²⁺ATPase,GSH-Px activity and MDA content are determined by colorimetry.VIP,5-HT,cAMP,PKA content and protein expression are detected by ELISA and Western blot.Results:1.Observe the model state.The swimming endurance,weight and rectal temperature were significantly decreased compared with the control group?P<0.01?;compared with the anti-evidence group,the swimming endurance and weight were increased?P<0.05?,and the rectal temperature increased dramatically?P<0.01?.2.Observe the morphological changes of gastric fundus and ileum mucosa in rats by HE staining.In the control group,the structure of ileal mucosa was intact and clear,the epithelial cells were tightly arranged and integrated.In the model group,the ileal mucosa became thin,and the villus became shorter or disappeared with epithelial cells.Necrosis and loss,interstitial blood vessels dilated and congested,inflammatory cell infiltration;compared with the control group,the structure of ileal mucosa was thicker,the villus was longer,the epithelial cells were with consistent arrangement,less necrosis and shedding,and less inflammatory cell infiltration in the anti-evidence group.3.Detecte the respiratory chain complexes I and IV,Na⁺-K⁺ATPase,Ca²⁺-Mg²⁺ATPase,GSH-Px activity,MDA content in ileum tissue with Colorimetric assay.In the model group,the activities of respiratory chain complexes I and IV,Na⁺-K⁺ATPase,Ca²⁺-Mg²⁺ATPase,and GSH-Px decreased rapidly and the MDA content dramatically increased?P<0.01,P<0.05?when it was compared with the control group.In the anti-evidence group,the activity of respiratory chain complexes I and IV,Na⁺-K⁺ATPase,Ca²⁺-Mg²⁺ATPase and GSH-Px markedly rose and the content of MDA was dropped?P<0.01,P<0.05?when it was compared with the model group.4.Detecte the VIP,5-HT,cAMP,PKA in ileum by ELISA In the model group,The content of VIP and 5-HT in the ileum of the model group grew considerably?P<0.01?when it was compared with the control group;In the anti-evidence group,the content of cAMP,PKA levels declined?P<0.01?,The content of VIP in the ileal tissue of the anti-evidence group declined?P<0.05?,5-HT was more marked?P<0.01?,cAMP,PKA levels climbed?P<0.01?when it was compared with the model group.5.Detecte the VIP,5-HT,cAMP,PKA protein expression in ileum by Western blot.In the model group,the expression of VIP protein in the ileum of the model group increased ?P<0.01?and 5-HT also grew?P<0.05?.The cAMP and PKA protein expression fell sharply?P<0.01?when it was compared with the control group.In the anti-evidence group,the expression of VIP and 5-HT in ileal tissue of the anti-evidence group dropped?P<0.05?,cAMP protein increased exponentially?P<0.01?,and PKA protein increased?P<0.05?when t was compared with the model group.Conclusions:1.The structural damage of ileal mucosal epithelium affects food digestion,nutrient absorption and transport,which is one of the pathological morphological foundations of spleen-qi deficiency syndrome.2.Intestinal mucosal injury,increased body lipid peroxidation,decreased mitochondrial synthesis energy function,impeded ion transport,affected signal transduction,disturbed gastrointestinal hormone,which is the pathological mechanism of spleen loss and clear spleen transport.3.the method that it invigorates Spleen and qi can restore ileal mucosal tissue morphology,restores normal cellular functions,increases body's antioxidant capacity and mitochondrial energy synthesis,enhances ion transport,strengthens cyclic adenosine monophosphate regulation,and improves gastrointestinal hormone secretion.It is one of the mechanisms of the efficacy of Spleen Qi deficiency syndrome.