The Effect Of The Method Of Replenishing Qi, Invigorating The Spleen, Removing Phlegm And Removing Blood Stasis, On AQPs In Kidney Tissue Of AS Bama Pigs With Spleen Deficiency And Phlegm And Its Mechanism

Purpose:To observe the changes of kidney water channel protein and cAMP/PKA signal pathway with the spleen dificiency and phlegm atherosclerosis bana mini-pig model and effect of the replenishing spleen on the model.Material and method:The part of literature research:Through collecting 47 ancient literatures from Qin and Han dynasty to Ming and qing dynasty,and more than 500 modern literatures.Make a summary that evolution of the etiology and pathogenesis,syndrome and treatment of thoracic obstruction in all ages.This raises syndrome of intermin-gled phlegm and blood stasis is one of the important syndrome type in cornary heart disease,spleen deficiency,phlegm and blood stasis is one of the main treatment methods for coronary heart disease.Meanwhile,it also raises the change of aquaporins in renal tissue probably is an index of dampness and blood stasis of spleen deficiency and phlegm stasis.It explain that the treatment of arteriosclerosis caused by spleen deficiency,phlegm and blood stasis is related to aquaporins in renal tissue and regulation of aquaporins in renal tissue by spleen governing transportation.The part of experimental:Fifteen bama minipigs were randomly divided into normal group,model group(spleen dificiency and phlegm atherosclerosis)and replenishing spleen group,replenishing spleen eliminating phlegm and resolving stasis group with five in each.The spleen dificiency and phlegm atherosclerosis model were instant by high fat diet combined tied by running after balloon injury in the coronary artery.The bama minipigs of normal group were fed normal basic diet for 48 weeks,model group were fed high fat and high calorie fodder for 48 weeks.The bana minipigs of replenishing spleen group and replenishing spleen eliminating phlegm and resolving stasis group were gived replenishing spleen recipe on the basis of the method on model group after 24 weeks.Observe content of potassium,sodium,chlorine,urea nitrogen,creatinine in serum.Study about regulatory hormone of respiratoy metabolism aldosterone(ALD),atrial natriuretic peptide(ANP),arginine vasopressin(AVP),neurotensin(NT).The pathological changes of kidney tissue were observed.Cyclic adenosine monophosphate(cAMP),Na~+-K~+-ATP enzyme of kidney tissue and the protein expression of aquaporins(AQP)1,2,3,phosphorylation protein kinase A(p-PKA),phosphorylated cAMP-response element binding protein(p-CREB)in the minipigs kidney after 48weeks.Results:1.Literature research suggests that the main syndromes of coronary heart disease nowadays is intermin-gled phlegm and phlegm and blood stasis,the main treatment methods of coronary heart disease is spleen deficiency,phlegm and blood stasis.Phlegm dampness syndrome in spleen deficiency,phlegm and blood stasis is related to aquaporins in renal tissue,aquaporins in renal tissue are closely related to metabolism of water and salt in vivo and regulatory ormone.Treatment of coronary heart disease of phlegm turbidity due to spleen asthenia from spleen deficiency and phlegm stasis may be related to the above mechanisms.2.1 Effect of the method for replenishing spleen eliminating phlegm and resolving stasis on potassium,sodium,chlorine,urea nitrogen,creatinine in serum of spleen dificiency and hlegm AS model.Compare with normal group,it is no change that potassium,sodium,chlorine,urea nitrogen,creatinine in serum in model group(P>0.05).About potassium,sodium,chlorine,urea itrogen,creatinine in serum,replenishing spleen group and replenishing spleen eliminating phlegm and resolving stasis group is no significant difference between them(P>0.05).2.2 The effect on method for replenishing spleen eliminating phlegm and resolving stasis for respiratoy metabolism and hormone level in spleen dificiency and phlegm AS.Compare with normal group,regulatory hormone of respiratoy metabolism ANP,AVP,NT were improved(P<0.05),and ALD a rising tendency(P>0.05)in model group.Compare with model,replenishing sleen eliminating phlegm and resolving stasis could reduce ANP,NT(P<0.05),AVP,ALD(P>0.05),and AVP is decreased in replenishing spleen group(P<0.05).Compare with replenishing spleen group,replenishing sleen eliminating phlegm and resolving stasis decrese ANP?NT(P<0.05).2.3 Effect of the method for replenishing spleen eliminating phlegm and resolving stasis on pathological changes of spleen dificiency and phlegm AS bama minipigs tissue.Compare with normal group,glomerular and renal tubular injury of kidney tissue were observed in model group.In normal group,glomerular structure clearly,lacuna of the renal capsule and the renal tubules were obvious.In model group,the glomerular was swollen,the renal capsule and the renal tubules became narrow.The renal tubules cells of model group arranged in disorder and swelling,casts were showed in collecting tubules.In replenishing spleen group,the renal capsule and the renal tubule showed the lacuna,but narrower than the normal group,glomerular swelling was better than that of model group,renal tubular disorder was not better.In replenishing spleen group,although lacuna of the renal tubular became ider,there was obvious damage in renal tubules and infiltration of inflammatory cells.Compare with model group,the renal capsule and the renal tubule of replenishing spleen eliminating phlegm and resolving stasis group showed the lacuna,also narrower than the normal group,glomerular swelling and renal tubular disorder was better than that of model group,no infiltration of inflammatory cells.glomerular swelling and renal tubular disorder of replenishing spleen eliminating phlegm and resolving stasis group is less than of eplenishing spleen group.2.4 Effect of replenishing spleen eliminating phlegm and resolving stasis on Na~+-K~+-ATP enzyme of spleen dificiency and phlegm AS bama minipigs tissue.Compare with normal group,Na~+-K~+-ATP enzyme was decreased(P<0.05).Compare with model group,replenishing spleen group improved Na~+-K~+-ATP enzyme in the kidney.P<0.05).Compare with model group,replenishing spleen group increased Na~+-K~+-ATP enzyme in the kidney(P<0.05).Compare with replenishing spleen eliminating phlegm and resolving tasis group,replenishing spleen group increased Na~+-K~+-ATP enzyme in the kidney better(P<0.05).2.5 The method for replenishing spleen eliminating phlegm and resolving stasis had impact on expression of AQP1,2,3 of spleen dificiency and phlegm AS bama minipigs tissue.Observation of immunohistochemical method indicated that expression of AQP1,2,3 in kidney of model group reduced compared with normal group(P<0.05),Compared with model group,replenishing spleen group increased expression of AQP1,2,3 in the bama mini-pigs idney(P<0.05).eplenishing spleen increased expressin of AQP1,2 better than of replenishing spleen eliminating phlegm and resolving stasis group(P<0.05).replenishing spleen liminating phlegm and resolving stasis group increased expressin of AQP3 better than of eplenishing spleen(P<0.05).Observation of westen-blotting method indicated that expression of AQP1?2?3 reduced(P<0.05)Compared with normal group.Compared with model group,replenishing spleen group increased expression of AQP1?2?3 in the bama mini-pigs kidney(P<0.05).replenishing spleen eliminating phlegm and resolving stasis group increased AQP3 in bana-pigs kidney(P<0.05),In replenishing spleen eliminating phlegm and resolving stasis group,the expressin of AQP3 increased(P<0.05),and the expressin of AQP1?2were increased(P>0.05).Expression of AQP1?2 in the bama mini-pigs kidney of replenishing spleen increased better than eliminating phlegm and resolving stasis group,but expression of AQP2 have bvious different(P<0.05).Observation of immunohistochemical and westen-blotting method indicated that xpression of AQP1?2?3 in the model bama mini-pigs kidney increased compared with ormal group,expression of AQP1?2?3 in the bama mini-pigs kidney of replenishing spleen increased better than eliminating phlegm and resolving stasis group,eplenishing spleen ncreased expressin of AQP1,2 better than of replenishing spleen eliminating phlegm and resolving stasis group.replenishing spleen eliminating phlegm and resolving stasis group increased expressin of AQP3 better than of eplenishing spleen.2.6 Effect of the method for replenishing spleen eliminating phlegm and resolving stasis on cAMP/PKA/CREB pathway of spleen difficiency and phlegm AS bama minipigs tissue.Compared with normal group,in model group cAMP?p-PKA?p-CREB reduced P<0.05).Compared with model group,the two drug administration groups could increase expression of cAMP,p-PKA,especially replenishing spleen group had statistical significance(P<0.05)in expression of cAMP,p-PKA,and replenishing spleen eliminating phlegm and resolving stasis group had no statistical significance in expression of cAMP,p-PKA(P>0.05).Conclusion:1.Literature research suggests that phlegm dampness syndrome in spleen deficiency,phlegm and blood stasis is related to metabolism of water and salt and regulatory mormone.The occurrence of coronary heart disease of intermin-gled phlegm and phlegm and blood stasis probably is closely related to metabolism of water and salt in vivo and regulatory hormone,treatment of coronary heart disease of phlegm turbidity due to spleen asthenia may be related to the above mechanis2.The part of experimental indicates that there are variation of AQPs and Liquid water metabolic hormones in the model of spleen dificiency and phlegm AS bama inipigs,treatment through replenishing spleen eliminating phlegm and resolving stasis could get well them2.1 There is no significant change about water and ion disorder and improving the serum of creatinine,urea nitrogen in the model of spleen dificiency and phlegm AS bama minipigs.However,ANP,AVP,NT were increased in model group.The method for replenishing spleen eliminating phlegm and resolving stasis could recover the unusual respiratoy metabolism and hormone level(Sijunzi decoction of the method for replenishing spleen has no obvious effect,and the method for replenishing spleen eliminating phlegm and resolving stasis is better than it.2.2 Spleen dificiency andphlegm atherosclerosis model group show glomerular and renal ubular swollen.replenishing spleen and replenishing spleen eliminating phlegm and resolving stasis could recover function,and the protective effect of replenishing spleen eliminating hlegm and resolving stasis is better than replenishing spleen.2.3 Spleen dificiency andphlegm atherosclerosis model group decrease of AQP1,2,3 and Na~+-K~+-ATP enzyme,methods for replenishing spleen and replenishing spleen eliminating phlegm and resolving stasis can improve the expression of AQP1,2,3.methods forreplenishing spleen could increase Na~+-K~+-ATP enzyme.2.4 In spleen dificiency andphlegm atherosclerosis model group,cAMP/PKA pathway was reduced.It indicates reduced expression of AQP1,2,3 related to cAMP/PKA pathway.Replenishing spleen and replenishing spleen eliminating phlegm group and resolving stasis group increased cAMP/PKA pathway and AQP1,2,3 in kidney,it shows the methods for replenishing spleen and replenishing spleen eliminating phlegm and resolving stasis improve expression of AQP1,2,3 related cAMP/PKA pathway.2.5 The ANP,AVP,NT in blood and cAMP/PKA,AQP1,2,3 in kidney of bama-minipigs model increased indicates variation of water-electrolyte metabolism hormone and AMP/PKA passway related with water-electrolyte metabolism in the model of spleen ificiency and phlegm atherosclerosis bana mini-pig model.Method of replenishing spleen liminating phlegm and resolving stasis regulated water-electrolyte metabolism hormone and kidney injury better than method of replenishing spleen,and method of replenishing spleen regulated cAMP/PKA/AQP passway better than the method of replenishing spleen liminating phlegm and resolving stasis.