Research Of The Function And The Mechanism By The Method Of Eliminating Phlegm And Removing Stasis To Prevent And Treat The Renal Fibrosis

ObjectiveThis study based on the theory of "the phlegm ties stasis mutually" is the pathogenesis key which the renal fibrosis occurs.Uses the clinical method which unifies with the animal experimentation, observes and discusses the clinical effect by the method of eliminating phlegm and removing stasis to prevent the renal fibrosis in renal treatment patients. And further discusses the function and the mechanism by the method of eliminating phlegm and removing stasis to prevent the big mouse model of renal fibrosis from TGF-β/Smads signal passage. Comprehensive evaluation of the clinical efficacy by use of eliminating phlegm and removing stasis to prevent and treat the renal fibrosis, and provide a new theoretical basis and methods of Chinese medicine for the prevention of renal fibrosis in renal treatment patients.MethodsThrough the collection of our hospital cases met the inclusion criteria of the renal treatment patients from January2011to January2012, We observe the change in patients on of clinical symptoms and laboratory parameters such as serum creatinine, urea nitrogen, albumin, blood lipids,blood analysis, liver filament four items and the level of TGF-β1,TIMP-1expression in renal fluid of the impact of herbs which can eliminate phlegm and remove stasis. We also can objective evaluate herbs clinical efficacy of prevention and treatment of renal fibrosis. Through establishing the big mouse model of renal flbrosis, observation the mouse model renal structure's histology change after the intervention of herbs which can eliminate phlegm and remove stasis. To expound the validity of herbs to oppress the big mouse model renal fibrosis. And applies the indirect immunity fluorescence method to determine the protein expression of each group of big mouse somatopleura renal such as TGF-β1, p-Smad2/3and α-SMA, E-cadherin and the p-Smad7, to discuss relations of phlegm reduction removes extravasted blood the traditional Chinese medicine to oppress the renal fibrosis with the regulative TGF-β/Smad signal passage's, promulgates initially the function mechanism.Result(一)The clinical research of prevention renal fibrosis by the law phlegm reduction and removal extravasted blood1,Phlegm and blood stasis influnce syndrome score of Chinese medicineAfter the treatment the improvement function of the treatment group clinical symptoms such as the facial color is gloomy, the flesh armor is wrong, the body numb or the stabbing pain, the feeling of heaviness in the limbs is sleepy, wan abdomen puffiness, in the mouth sticks greasily, dry mouth do not want to drink, losses of appetite, stool viscosity surpasses the control group obviously (P<0.05), which show phlegm reduction removes extravasted blood the traditional Chinese medicine to improve the symptoms, the treatment group better than the control group.2,Phlegm and blood stasis related indicators of the impact of renal fibrosisAfter treatment the treatment group TGF-β1levels effluent decreased significantly, compared with before treatment, there was significant difference (P<0.05);the control group compared with pre-treatment showed no significant difference (P>0.05);After treatment, comparison between the two groups, the treatment group TGF-β1levels renal treatment effluent decreased was significantly higher (P <0.01). After treatment the treatment group TIMP-1levels renal treatment effluent were significantly decreased, compared with before treatment, there was significant difference (P<0.05); the control group compared with pre-treatment showed no significant difference (P>0.05); After treatment, comparison between the two groups, the treatment group factor of decline in effluent TIMP-1levels were significantly higher (P<0.01). Indicators reflect the degree of fibrosis:hepatic-fibrosis four items, treatment and control groups before treatment compared after treatment were decreased,but only the treatment group has statistically significant difference (P<0.05), no statistical difference between the control group significance (P>0.05);After treatment between the two groups was statistically significant difference (P<0.01); showed that patients treated with phlegm reduction and stasis removal herbs renal treatment on hepatic-fibrosis four items have some improvement. In conclusion, the results suggest that phlegm and blood stasis can significantly reduce the level of renal fibrosis-related indicators, play antagonistic renal fibrosis, and protect the renal function.3,Phlegm and blood stasis effect on laboratory parametersTreatment group after treatment with phlegm and blood stasis, the improvement of creatinine, urea nitrogen, total cholesterol, triglycerides, low density lipoprotein, fibrinogen better than the control group (P<0.05), indicating that the phlegm and blood stasis in improving renal functi on, blood lipid and cogulation have a good effect.4,Phlegm and blood stasis indicators of drug safetyTreatment group during the medication, the symptoms do not appear all kinds of adverse reactions, and reflecting the heart and liver function indicators of CK-MB, AST, ALT, before and after treatment, the difference was not statistically significant (P>0.05), showed that patients treated with phlegm and blood stasis, on cardiac function and liver no functional effect, taking safety.(二)Experimental study of phlegm and blood stasis to Prevent and Treat the renal fibrosis1,Observation renal tissue of rats and comparison dense layer thickness Control group, HE staining showed thin and dense renal smooth, flat layer of mesothelial cells covering intact, thin basement membrane between the skin and connective tissue attached to the skin, between the collagen fibers had no significant deposition, capillary dilatation and congestion-free, non-inflammatory cell infiltration. Model group was significantly thicker renal, mesothelial cell loss, spindle fiber cells, fibroblasts, generated by multi-layer, between the loose skin matrix, a large number of collagen fiber deposition, capillary dilatation and congestion, accompanied by inflammatory cell infiltration, suggesting that renal fibrosis model success. The western medicine group and the morphological changes of rat renal have different levels of mitigation to reduce, the level of medicine in high dose group significantly. The group compared the thickness of dense layer parietal renal, renal thickening model rats in which the most obvious level, compared with the control group were significantly different (P<0.01); Chinese medicine high dose group and low dose group and western medicine group were significantly different (P<0.05); western medicine group and low dose group, significant differences (P<0.05). Phlegm and blood stasis can be effectively shown to improve renal fibrosis pathological changes, especially in the high dose of Chinese medicine.2,Comparison of immunohistochemistryImmunofluorescence showed that Smad2/3was mainly expressed in the nucleus, Smad7, α-SMA and TGF-β1expressed mainly in the cytoplasm. Control group, Smad2/3, TGF-β1, α-SMA, Smad7expression is low, and the expression of E-cadherin is obvious; Model group Smad2/3, Smad7, TGF-β1and α-SMA expression in a high, E-cadherin expression is very low; the two groups there was a significant difference(P<0.01); In the high group and low group and western medicine group Smad2/3, TGF-β1and α-SMA expression compared with model group decreased, but Smad7, E-cadherin expression compared with model group increased, compared with model group, three groups were significantly different (P<0.05); in the high group Smad2/3, TGF-β1and α-SMA expression than those in the low group and western medicine group significantly reduced. In the high group were compared with the latter two groups were significantly different(P<0.05); In the high group of Smad7, E-cadherin expression than those in the low group and western medicine group was significantly increased. In the high group were compared with the latter two groups were significantly differences (P<0.05); Western medicine group Smad2/3, TGF-β1and α-SMA expression decreased than those in the low group, Smad7, E-cadherin expression was increased by lower than those in the two groups were significantly different(P<0.05).Conclusion1,Through January2011to January2012,40patients met the criteria of the standard treatment of clinical studies suggest that: phlegm and blood stasis can improve clinical symptoms of patients and treatment effection.2,Blood stasis and phlegm can significantly reduce the level of relevant indicator of renal fibrosis (liver fiber four, renal fluid TGF-β1, TIMP-1factor), can protect the structural integrity of renal morphology and reduce the rat model of renal fibrosis, play the role of renal fibrosis antagonist. This effect was dose-effect relationship, the best high-dose group.Phlegm and blood stasis initially showed better prevention and treatment of renal fibrosis. Through5/6nephrectomy rats application of caused by renal fibrosis were found: phlegm and blood stasis on the impact of TGF-β/Smad signaling pathway possibly by increased TGF-β signaling inhibition protein expression of Smad7and inhibit TGF-β receptor signaling protein regulated (Smad2/3) activation, prevents renal fibrosis in animal models of renal mesothelial cell transdifferentiation occurs, thereby preventing or reducing the incidence and progression of renal fibrosis, Protecting the renal function.