Study On The Relationship Between Of The Effect Of Rehmannia Root In Regulating AQP2 Expression And Improving The Micro Inflammatory State(Heat Depression)

BackgroundDiabetes mellitus(DM) is a group of metabolic diseases characterized by chronic elevated of glucose level in serum, which is caused by abnormal insulin secretion and(or) effect, which threat to human health seriously. DM patients with long-term high blood sugar disease gradual progression to DM late, the human body produce a series of complications, and ultimately lead to a chronic change in various tissues and organs, in the eye, kidney, heart, blood vessels and so on, even the function decline and failure in multiple organ. Diabetic nephropathy(DN), which is one of the most important risk factors in the end stage of diabetes, is becoming one of the most important risk factors gradually;Recent studies have suggested that the presence of Micro inflammatory state in patients with type 2 diabetes mellitus(T2DM) is a process of innate immunity and low degree of chronic inflammatory disease. Pick et al. think that diabetes is a kind of natural immunity and disease with low inflammatory, and it is suggested that T2 DM has the acute phase response hypothesis, which is mediated by cytokines. The micro inflammatory state which so-called is different from pathogenic microorganisms caused by infection, is also different in systemic inflammatory response syndrome(systemic inflammatory response syndrome(SIRS),is affected by many factors(microbial and endotoxin, chemicals, immune complexes, complement, etc.) as a result of stimulation, to proinflammatory cytokine release inflammatory reaction in the center. The main clinical manifestations were systemic circulatory filling in low level and sustained inflammatory factors increased, it is contribute to the occurrence and development of diabetes and related complications. Recently, the role of inflammatory cytokines in the occurrence and development of DN has been gradually concerned. The micro inflammatory state and immune function disorder of the body lasting in the long term,may be one of the most important pathogenesis in DN. It could cause the development and deterioration in DN, it can also promote the occurrence of various complications in cardiovascular disease, malnutrition, nerve degeneration,and so on; is one of the main factors which can affect the prognosis in patients with DM; therefore, it could be an important clinical significance in strengthen the study in the micro inflammatory state of the pathogenesis and in treatment of DN.Duration of DM with the passing of time, persistently elevated inflammatory factor, patient tend to the existence of blood stasis phenomenon, complexion is often Leahey dull, the tongue is dark purple. According to the principle of cooling and activating blood treating diabetes, to improve the micro inflammatory state in patients with DM.Rehmannia glutinosa for the Scrophulariaceae plant, is a kind of traditional Chinese medicine, commonly used in its root, the main producing in Henan Huaiqing, as a representative drug of "cooling blood and activating blood circulation", was originally recorded in the Shennong Bencao Jing ", on the properties of the introduced to:" the main fold down the vast tendons, injury, by blood arthralgia, fill the bone marrow, muscles, as soup in addition to cold and it can heat accumulation and removing stagnation, living, especially good ". Modern research has confirmed that Huang can reduce blood sugar, diabetes control, has a good effect on DM.For further study of the therapeutic effect and mechanism of Rehmannia glutinosa on micro inflammation, we establish a diabetic rats model with the manner of high glucose and high fat diet combine with intraperitoneal injection of streptozotocin(STZ) to damage the islet cells in SD. Treatment with Salvia miltiorrhiza Bge, Rehmannia glutinosa, in method of gastric lavage respectively, once a day for 60 days. Monitoring the blood glucose with blood glucose meter, Detect the concentration of high sensitive C reactive protein(hs CRP), interleukin- 6(IL-6) and tumor necrosis factor alpha(TNF-?) in serum by ELLSA; research to observe the improving effect of Rehmannia glutinosa on blood glucose and diabetes micro inflammatory state by means of immunohistochemistry, pathology,to investigate the effect of diabetes on micro inflammatory state in effect and its mechanism, study the regulation effect of Rehmannia glutinosa on the expression of AQP2 in kidney of diabetic rats. The results of the study are as follows:Experiment 1: The effect of Rehmannia Root on the micro inflammation state in diabetic ratsObjective:1. To establishment a diabetic rat model by injection STZ intraperitoneal with high glucose and high fat diet.2. There are micro inflammatory state in Diabetes, To study whether the Rehmannia Root has any improvement in micro inflammation state.3. To study whether the Rehmannia Root has any improvemen effect on renal tissue injury in diabetic rats.Method:60 male SD rats were divided into two groups randomly, 10 in normal group, and 50 in the model group, and model group were fed with high glucose and high fat diet and intraperitoneal injection(40mg/kg)(STZ). Model rats were divided into diabetic model group, western medicine group, Salvia miltiorrhiza group, Rehmannia group. Observation of rats weight, spirit and hair, limbs activities basic state, regular inspection of rat blood and urine glucose confirm diabetic model was successfully built; ELISA assay was used to detect the serum interleukin-6(IL-6) and tumor necrosis factor alpha(TNF alpha), high sensitive C reactive protein(hs CRP) content. Hematoxylin and eosin(HE) staining to observe the pathologic changes of kidneys.Results:The application of high glucose and high fat diet plus streptozotocin(STZ) intraperitoneal injection of the diabetic rats model was successful; rehmannia can significantly reduce blood glucose levels in diabetic rats,it can reduce TNF-?, Hs-CRP, IL-6 content in serum of rats; kidney tissue HE staining can be seen in normal glomerular structure rats clear and intact basement membrane, no renal interstitial inflammatory cell infiltration; glomerular hypertrophy of diabetic rats in the model group, the ball inside erythrocyte sedimentation, mesangial area widened, and the proliferation of mesangial cells, renal tubular epithelial cell swelling and shedding, vacuolar degeneration; glomerular Rehmannia rats can be seen in the trace of erythrocyte sedimentation, no epithelial cell shedding, a small amount of inflammatory cell infiltration in the stroma.Conclusion:1. The diabetic rat model was successfully established by intraperitoneal injection with STZ and high glucose and high fat diet;2. Rehmannia can effectively control blood glucose in diabetic rats and improve the micro inflammatory state of diabetic rats.3. Rehmannia can improve the degree of renal damage in diabetic rats effectively.Experiment two: The effect of Rehmannia on renal aquaporins regulate the expression of AQP2 in diabetic rats.Objective:To discussing the essence effect of " Cooling blood and activating blood circulation drugs " in Rehmannia through regulating the AQP2 expression of digitalis for the renal tissue of diabetic rats.Methods:the model and grouping method is the same as above. The expression of AQP2 in kidney tissue of rats in each group was analyzed by blot Western, and the expression of AQP2 was detected by immunohistochemical method.Results:the expression of AQP2: in the kidney tissue, compared with the model group, the expression of AQP2 was significantly lower than that of the model group, the difference was statistically significant.Conclusion:Rehmannia has inhibitory effects on AQP2 in renal tissue of diabetic rats.Summary:1.Confirmed the method of fed with high glucose and high fat diet,intraperitoneal injection with STZ can established diabetic rat model successfully;2.Rehmannia Root can significantly reduce the blood glucose of diabetic rats.3.Cooling blood and activating blood circulation drugs Rehmannia can reduce the content of blood TNF-?, hs-CRP, IL-6, can improve diabetic micro inflammatory state effectively.4. The damage of the kidney tissue caused by diabetes has a significant improvement by Rehmannia Root.5. The effect of AQP2 on the expression of in renal tissue of diabetic rats could be decreased, and the concentration of urinary AQP2 in rats could be decreased, which might be one of the modern mechanisms of "promoting blood circulation".