| The viewpoint of "the blood stasis can form edema,so blood is like water to someextent", stated from 《synopsis of the golden chamber》of Zhang ZhongJing,suggeststhat the relationship of blood and water is transforming each other and similar functionsin physiology,a close association between blood stagnation and water retention.Themethod of "promoting blood circulation and alleviate water retention" based on the point of "Blood Syndrome Theory".The method reflects the two effects of "promoting bloodinvigorates circulation of water and alleviating water retention benefits promoting blood",but the two effects have the difference of "alleviating edema and diuresis ". So we wantto explore the nature of " activating blood circulation and promoting diuresis " fromclinical practice and pharmacology,"detumescence and diuresis" should be consideredas the breakthrough point.AQPs is a kind of cytomembrane glycoprotein, the molecular weight about30kd,atetramer in cytomembrane, relating to the water permeability (absorption, secretion,transport) to maintain extra-and intra-cellular water balance.AQP1is widely distributedin the body tissue cells, especially in epithelial cells and endothelial cells in connectionwith the water absorption and secretion.AQP1is the main Aquaporin in Erythrocytemembrane that regulate water transport between blood and intracellular in erythrocyte,improving erythrocyte deformation ability,and having some connection with "bloodstasis". So AQP1is associated with blood stasis and tissue edema.AQP1in mainlyconcentrated in the proximal tubules, collecting tubule and thin segment in kidney,AQP2expressed mainly in distal tubule and principle cells membrane in collectingtubule, both are involved in water absorption and urine concentration, and the functionof AQP1and AQP2anomaly is associated with "diuresis". So we think that AQP1andAQP2is the critical molecule that reveal the part nature of "activating blood circulationand promoting diuresis ".Salvia miltiorrhiza has the effect of promoting blood circulation to dissipate bloodstasis detumescence and odynolysis. Modern research on the function of "promotingblood circulation to dissipate blood stasis" reflects its lowering blood viscosity,inhibiting platelet aggregation, thrombosis and improving microcirculation and otherpharmacological effects,"detumescence and odynolysis" embodies its reducing vascularendothelial cell damage, reducing the vascular permeability, reducing vasogenicedema.Therefore,we explore to reveal the nature of "promoting blood circulation andalleviate water retention" by salvia miltiorrhiza regulating the characteristic expressionof AQPs. Experiment1: Salvia miltiorrhiza regulated AQP1expression in local muscleand kidney tissueObjective:discussing the method of promoting blood circulation and alleviatingedema. Methods:SPF50SD rats were divided into two groups at random: contrastgroup with10and model group with40.The rats model were established by quantitativeblow.40model group rats were divided into blank control group, salvia miltiorrhizagroup, inhibiting group and intervention group.The pathological change of injuredkidneys and muscles by HE staining,the microstructure changes of injured kidneys andmuscles by electron microscope,the changes of dry and wet ratio of muscles andkidneys,the AQP1expression in kidneys and muscles by ICC and Westren-blotmethods. To explore how to produce the rat model of blood stasis due to trauma,andstudy the pathological features of blood stasis and fluid retention syndrome in themodel.Results:1.Compared with contrast group, rats in model group manifested thesymptoms of spiritual malaise, eating less obviously, daily water increasing, more urineoutput and the injured hind-limb existing purple-red swell and limp. Salvia miltiorrhizagroup rats hind limbs swelling significantly reduce compared with blank control group,inhibiting group no significant relief.2.Pathological results:Because of inflammationdue to traumatic blood stasis, massive capillary hyperplasia and a small amount ofinflammatory cells such as lymphocytes, plasma cells,could be observed around theinjured muscle fibres in blank control group. And in the kidney tissue can be foundthat:glomerular capsule have dilatation and blood stasis to different degrees,VEC’slumen is narrowed and erythrocyte aggregation,and the epithelial cells of renal tubuleswelled. Salvia miltiorrhiza group can find blood stasis conditions improved in localemuscle subcutaneous, inflammatory cells disappeard, muscle fiber structure closed tocontrast group. Kidney renal tubular-interstitial lesion is improved, a few interstitialcapillary mild hyperemia.Inhibition group can be found in a small amount ofinflammatory cells.3. Electron microscopy results:In the muscle tissue of the modelgroup we can find that nucleus transfer,mitochondria swell vacuolar degenerate andinterstitial capillary congest. In the kidney tissue the epithelial cells of renal tubules demonstrated vacuolation and had some protein droplets,vacuolar degenerate,podocytevacuolated,and hyperemia of peritubular and interstitial capillary can be found. Salviamiltiorrhiza group can be found mitochondrial proliferation, some swelling in musclefiber, glomerular epithelial cell swelling, mild swelling of mitochondria, endoplasmicreticulum minor expansion.Inhibiting group closed to blank control group.4.Theexpression of AQP1:Compared with the contast group,the AQP1’expression wasincreased in the muscle tissue（P<0.05) and reduced in the kidney tissue（P<0.05).Compared with the blank control group, the AQP1’expression was increased（P<0.05) inSalvia miltiorrhiza group.Conclusion:1.In blood stasis model,Salvia miltiorrhiza can obviously alleviate thesymptoms of stasis,swelling in the injured muscle and renal tissues and increase theexpression of AQP1,which shows that the effect of "promoting blood circulation andalleviating edema" about Salvia miltiorrhiza may be through increasing the expression ofAQP1.2.The occurrence of blood stasis may be related with the VEC’s damageclosely.3.The decrease of RBC’s deformation may be associated with the occurrence ofblood stasis and the AQP1is the main Aquaporin on RBC.Based on the above twofacts,we speculated that "promoting blood circulation and alleviating edema" may alsohas relationship with AQP1on RBC, but the specific mechanism is still an openquestion.Experiment2:The effect of salvia miltiorrhiza to the expression of AQPs inkidneyObjective:to reveal the nature of "alleviating water retention " by the influence of salviamiltiorrhiza.Methods:24hours’ urine volume of all rats with metabolism cages,thelevels of urine AQP2with Enzyme linked ELASA,the AQP1and AQP2expression inkidneys by ICC and Westren blot methods.Results: The levels of urine AQP2in blankcontrol group level was higher than normal group (P <0.05), Salvia miltiorrhiza group’swas lower (P <0.05).2. AQP1, AQP2expressed: compared with blank control group,salvia miltiorrhiza group rats kidney AQP2expressed significantly decreased (P <0.05),AQP1expression increased significantly (P <0.05). Conclusions:1.Salvia miltiorrhiza can obviously alleviate renal tissue edema andincrease urine output in blood stasis model.2.Salvia miltiorrhiza can decrease theexpression of AQP2and increase the expression of AQP1,which reveals that the effect of"invigorating blood circulation and diuresis" may be related with the changes of AQP’sexpression.Summary:1.The method of " activating blood circulation and promoting diuresis " can be dividedinto the two subtype method:activating blood detumescence,invigorating bloodcirculation and diuresis.2.The effect of alleviating the edema of local muscles and renal tissue about Salviamiltiorrhiza may be through increasing the AQP1’s expression in VEC and renalproximal tubular cells and decreasing the AQP2’s expression in renal distal tubule andcollecting duct. So the method of "promoting blood circulation and alleviating waterretention"is associated with AQP1and AQP2.3.The ability of RBC’s deformation may be associated with the occurrence of bloodstasis and the AQP1is the main Aquaporin on RBC.So we speculates that salviamiltiorrhiza can increase the expression of AQP1on RBC membrane and improve itsability to transport water, thus reducing the excessive water in blood.4.AQP1and AQP2can be used as the key molecule for researching the two subtypemethods of "promoting blood circulation and alleviating water retention "... |
PDF Full Text Request