| Research background:Blood stasis is blood failed to timely discharge or disappear, stay in the body; or blood runs sluggish, product within the Zang and Fu, organs, tissues, loss of normal physiological function of the syndrome, which is considered as blood stasis syndrome.Traditional Chinese medicine theory is that the generation of blood stasis is closely related to the two factors of blood and vascular, 1. Nourishing blood and nourishing function of play are easily influenced by the factors of cold and heat, Qi stagnation, Qi deficiency, so the "Lingshu " has discussion of "xue huo and jing mai liu xing". In addition, traditional Chinese medicine has " jin and blood homologous ", " sweat and blood homologous ", " jing and blood homologous " point of views, Thinking they can alternate each other. Therefore, we think that blood stasis is to a certain extent accompanied by water and liquid stop.2. "Pulse, Vein blood of the house" the " Su Wen " was recorded Vessel is not only the blood of places and blood running channel, so pulse Tongli or not is whether the flow of blood is the premise and guarantee.Modern research confirmed microcirculation disturbance of microcirculation abnormality(blood), which can change the micro blood flow, blood "sticky", and showed the pathology of blood stasis; secondly, there are micro vascular endothelial cell injury(pulse), vascular endothelial cells(VEC) is tissue membrane, lining affixed to the intravascular side, close contact is an important part of the "pulse", and the blood, internal environment stable, tissue blood supply and metabolic regulation is very important. Vascular endothelial cell damage can lead to the changes in vascular permeability and plasma protein infiltrated into the tissue space, tissue fluid colloid osmotic pressure increased, resulting in "water product, blood stasis and" water product "as, the TCM"jin and blood homology "" blood sweat homologous "" Jingxue homologous "point of view coincide. And previous treatment of blood stasis syndrome focus in blood and ignore vein tract injury caused by water accumulation, so this topic proposed concept of "water and blood stasis", to create "blood-activating and water-disinhibiting" and can be divided into two aspects of "Huoxue Xiaozhong" and " blood-activating activating diuresis " is reflected in the lung and kidney tissues.Under the guidance of the theory "disease in vein, regulating blood, disease in blood, regulating collaterals" in the "Suwen ", VEC can be used as the effect site to improve microcirculation disturbance.AQP is cell membrane glycoprotein which is related with water permeability like Absorption, secretion, transport. It can maintain water balance inside and outside cells. AOP1 selectively distributes endothelial cells and epithelial cells etc. AQP1 was selectively distributed in endothelial cells(VEC) and epithelial cells, which were expressed in lung and kidney tissues.The test shows that AQP1 is great important to Angiogenesis and cell migration. And it is conducive to VEC repair.AQP1 can be the new index when cure microcirculation disturbance with activating blood and dieresis.Salvia miltiorrhiza, as a traditional blood activating drug, could be used to investigate whether Salvia miltiorrhiza could improve the microcirculation disturbance and regulatethe effect of AQP1 by protecting VEC, and validate vascular endothelial cells is an effective target to improve blood stasis.Part1: The protection of Salvia miltiorrhiza to vascular endothelial cell of acute microcirculation disturbance ratObjective: 1.Verify the model of blood stasis syndrome is successful: Injection of adrenaline hydrochloride and ice water stimulation lead to rats get acute microcirculation disturbance, be blood stasis syndrome. 2. Observe the specification change of blood stasis syndrome, verify Salvia miltiorrhiza can improve microcirculation disturbance by protecting endothelial cells.Methods: normal group 10 rats, the model group 40 rats, Rats in model group were injected with adrenalin hydrochloride in the abdominal skin and stimulated by ice water. When model group is successfully built, the 40 rats are divided into the normal group, the model group, western medicine control group, salvia miltiorrhiza low doses group, salvia miltiorrhiza middle doses group,and salvia miltiorrhiza higher doses group. Observe the basic status of each rats,sunch as their mind, hair, tongue color etc. Shave ear hair, detect the state of ear microvascular blood flow by microcirculation apparatus and blood viscosity and erythrocyte deformation capacity, to confirm whether microcirculation obstacle model is successful or not. The method of ELISA can test the content of ET-1 in the serum, P-Selection and VEGF.Results:Salvia miltiorrhiza can improve rats ear microcirculation disturbance obviously and reduce the blood viscosity of rats, it can reduce the ET-1、P-Selection、VEGF prominently.Conclusion: Salvia miltiorrhiza can improve rats acute microcirculation disturbance which caused by adrenaline hydrochloride and ice water stimulation, has relationship by protecting endothelial cells and validate vascular endothelial cells is an effective target for blood stasis syndrome.Part2: Salvia miltiorrhiza improve acute microcirculation disturbance rats who gets lung injury and the adjustment of lungs tissue AQP1.Objective: By salvia miltiorrhiza for the regulation of microcirculation of rat lung AQP1 expression, To validate Salvia miltiorrhiza by up-regulation the expression of AQP1 inlung tissue endothelial cells to reduce lung injury and pulmonary edema in rats with mild circulatory impairment.Method: Take the rat lung tissue test the parameters of pulmonary edema. lung wet dry ratio(W/D), and of the right lung /body weight ratio(R/B); HE staining observe each lung tissue pathology morphology; Using Western blot detection methods to analyze each rat lung tissue AQP1 expression; Using immunohistochemical detecting method to each group the AQP1 expression of lung tissue in rats.Results: the lung wet dry ratio and the ratio of the right lung /body weight tip: compared with normal group, model group increases significantly(p < 0.05) salvia miltiorrhiza dosage, Compared with the model group, the ratio of salvia miltiorrhiza with high dose is proved to lower(p < 0.05); HE staining results show: the model group has the change of acute lung injury, salvia miltiorrhiza has obvious effect and the structure of the lungs becomes normal with high dose; AQP1 expression shows: In the lung tissue, salvia miltiorrhiza of expression with low, medium and high dose is proved to increased, compared with model group, and increased with salvia miltiorrhiza dose increase obviously.Conclusion: salvia miltiorrhiza is used to raising lung endothelial cells on the expression of AQP1 to reduce the rats’ acute lung injury caused by adrenaline hydrochloride and alleviate edema,and verificate “blood stasis and water retention” is of “blood stasis syndrome”,which reflects the the method of activating blood detumescence.Part3: Salvia miltiorrhiza improve microcirculation disturbance rats who gets acute kidney injury and the adjustment of kidney tissue AQP1.Objective: Through observing the adjustment of salvia miltiorrhiza is to microcirculation disturbance rats kidney AQP1, To validate Salvia miltiorrhiza by up-regulation the expression of AQP1 to improve water metabolism disorder induced by renal injury in rats with microcirculation disturbance.Method: Observe the pathological morphological changes of each kidney tissue by the method of HE staining.Analysing the Kidney tissue AQP1 expression of each group rats by the detection method of Western blot. Detect the Kidney tissue AQP1 expression ofeach group rats by the method of immunohistochemistry.Results:HE staining result shows: Model groups has the kidney injury, salvia miltiorrhiza can improve the microcirculation disturbance obviously, and the more dosage, the more normal structure and functions of kidney; AQP1 shows: In the kidney tissues, different dosage of salvia miltiorrhiza can rasie the expression of model group, and the more dosage of salvia miltiorrhiza, the more obvious expression.Conclusion: Salvia miltiorrhiza can improve the renal injury in rats with microcirculation disturbance has the relationship of increasing AQP1, and reflect the importance of Activating blood circulation and promoting diuresis for blood stasis and water retentionSummary: 1. Verify the rats which stimulated by adrenaline hydrochloride and ice water swim get microcirculation disturbance. 2.Invigorate the circulation of blood and water flooding medicine salvia miltiorrhiza protect vascular endothelial cell by reduce the ET-1、P-selection、VEGF factor, to improve microcirculation disturbance validate vascular endothelial cells is an effective target for blood stasis syndrome.. 3. Maybe it has the relationship of protecting microvascular endothelial cell, when salvia miltiorrhiza can improve acute kidney injury and acute lung injury caused by acute microcirculatory disturbance. 4. Salvia miltiorrhiza improve microcirculation in rats with acute lung injury and the effect of pulmonary edema index results by up regulating the expression of AQP1 further verify the presence of water and blood stasis in "blood stasis", reflects the method of activating blood circulation and subsidence of a swelling. 5. The effect of Salvia miltiorrhiza on the prevention and treatment of water and fluid metabolism disorder caused by the injury of kidney by up regulating the expression of AQP1. It reflects the importance of Activating blood circulation and promoting diuresis for blood stasis and water retention... |
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