| Objective:Explore and improve the production of Qi-Yin deficiency and blood stasis syndrome of diabetic nephropathy(DN)animal models,and confirmed the protective effect of Tang-Shen-Kang to DN kidney of rats in the experimental point of view.Methods:Totally 50 healthy male SD rats were randomly divided into normal group 8and model 42 by randomized block method,then successful model rats were randomly divided into 4 group:Model group, Tang-Shen-Kang group, Benazepril group, disease-syndromeme model group(Qi-Yin deficiency and blood stasis animal model).Rats in each group pre-test blood glucose, 24 hours Urine Protein(24h UPr), was negative for the experiments.In addition to the eight rats of normal group, the other 42 rats are used to make DN animal models by triple law(surgery + high-sugar and high-fat diet +streptozotocin).After the modeling, the normal group, model group were fed a fixed animal feed and water.Tang-Shen-Kang group with 1.6g/(200g· d)Tang-Shen-Kang dissolved in warm water by gavage.Benazepril group with 0.3mg/(200g· d)benazepril dissolved in warm water by gavage.The rats fed once daily and 8 weeks of treatment.disease- syndromeme model group will make Qi-Yin deficiency and blood stasis DN animal models.After treatment, the rats in each group abdominal aortic blood and removal of the right kidney.After quickly weighed, the kidney will be fixed in neutral formalin.All the rats stay 24 h urine before surgery.Detecte ACR by radioimmunoassay;detecte 24 h UPr by biuret method; detecte BUN by enzyme-coupled rate method;detecte Scr by creatinine enzymatic; renal pathological changes were observed by HE, Masson + PASM.Results:1.TCM symptoms in rats, signs of change before and after reatment:Water conditions, mental status, daily activities were normal in normal group, steady increase in body weight of rats.After the model rats inject STZ 72 h, the performance of rats as polydipsia, polyphagia, polyuria, gradual weight loss, listlessness, activity slow,Unresponsive, coat haggard and so easy to take off.With time, part of rats appear a variety of DM complications gradually,Such as the body and tail of infection ulcers.Food intake and water intake in Tang-Shen-Kang group gradually increased, mental state is nomal, the body hair is brighter than model group, activity increased gradually,body weight is more stable, compared with the model group.Benazepril group and Tang-Shen-Kang group have similar performance.After the disease- syndromeme model group make successfully,the rats show restless, more drink, poor coat gloss,purple or green spots subcutaneous stasis, gradual weight loss.Normal rats no deaths, 5rats died during making models, 2 rats was died in model group, Tang-Shen-Kang group and benazepril group have died 1 rat after treatment, disease- syndromeme model group have died 3 rats in making Qi-Yin deficiency and blood stasis animal models. The cause of death may be fed, infection, biting each other,making disease-syndromeme model group and so on.2.Comparison of body weight of rats in each group and kidney hypertrophy index:Compared with the same period in normal group, the rest rats’ weight of the group were significantly reduced(P<0.01).The 2 weeks and 4 weeks of treatment, there were no significant difference between benazepril group and Tang-Shen-Kang group and model group(P>0.05).The 6 weeks and 8 weeks of treatment, Tang-Shen-Kang group and benazepril group compared with model group have significant difference(P<0.05 or P <0.01), Tang-Shen-Kang group have no significant difference compared with benazepril group(P> 0.05).Kidney hypertrophy index, compared with the normal group, the other groups were significantly increased(P <0.01). Compared with the model group, Tang-Shen-Kang group and benazepril group were decreased(P<0.01).Tang-Shen-Kang group and benazepril group have no significant difference(P>0.05).3.Renal function compares each group(BUN、Scr):By one way ANOVA, the rats BUN and Scr was no significant difference(P> 0.05).4.ACR and 24 hUPr compares in each group:compared with normal group,ACR and 24 h UPr in each group increased significantly, with a very significant difference(P<0.01). compared with model group, Tang-Shen-Kang group and benazepril group were decreased significantly in 4 weeks and 8 weeks treatment(P <0.01),Tang-Shen-Kang group have no significant difference with benazepril group(P> 0.05).8 weeks treatment compared with 4 weeks treatment have no significant difference in Tang-Shen-Kang group and benazepril group(P> 0.05),but have significant difference compared with no treatment(P< 0.05 or P <0.01).5.Renal pathological morphology:(1)renal tissue with a naked eye:the size of the rats’ right kidney is normal and the color is dark red in normal group.The size of the rats’ right kidney is significantly increased in model group and disease- syndromeme model group; Tang-Shen-Kang group and benazepril group of rats’ right kidney is bigger than the normal group, but smaller than the model group.(2)HE staining: The glomerular structure of normal group is clear. The glomerular balloon part stick together and cysts significantly expand in model group and disease- syndromeme model group.Renal capsule expand mildly in Tang-Shen-Kang group and benazepril group. Mesangial cells and endothelial cells in each group showed no significant increase.(3)Masson+PASM staining: The performance of glomerular basement membrane: the model group was significantly thicker than the normal group, the performance of the basic pathological changes in DN; and disease- syndromeme model group, more significantly thicker than the model group, may have relationship with longer duration as well as manufacturing syndromes model closely. Benazepril group and Tang-Shen-Kang group roughly equal,compared with the normal group slightly thickened, but significantly reduced compared with the model group. The performance of mesangial matrix: the model group increased significantly compared with the normal group; disease- syndromeme model group compared with the model group increased more significantly. Benazepril group and Tang-Shen-Kang group roughly equal, compared with the normal group increased slightly, but significantly reduced compared with the model group.Conclusions:1.Tang-Shen-Kang can improve the quality of life of DN animal models, reduce urinary protein of DN rats;2.Tang-Shen-Kang can improve the renal morbid pathology state of DN animal models;3.The production of Qi-Yin deficiency and blood stasis syndrome DN animal models is successful.Objective:By observing the clinical efficacy of the Tang-Shen-Kang on the DN Qi-Yin deficiency and blood stasis syndrome patients and the change of 24 h UPr and ACR before and after treatment, and from a clinical point of view confirmed that Tang-Shen-Kang could protect the kidney of DN patients of Qi-Yin deficiency and blood stasis.Methods:68 cases DN patients with Qi-Yin deficiency and blood stasis syndrome from June 2013 to January 2015 in Department of Nephrology and Endocrinology were randomly divided into treatmenn group(34 cases) and control group(34 cases).In the course of treatment lost five cases, two cases in the treatment group and three cases in the control group, the actual completion of 63 cases, and 30 cases of normal healthy persons as normal group.The two groups of patients were given comprehensive treatment to control diet, blood glucose and blood pressure; On the comprehensive treatment of the above,the treatment group mix in Tang-Shen-Kang,3 times a day, each time a bag, a course of eight weeks.Observe the efficacy of disease, symptom scores,ACR and 24 h UPr, kidney function, blood glucose, etc.And observe the adverse reactions. According to the experimental results,the statistically will be analyzed and the efficacy will be determined.Results:1.Comparison of the effect of clinical disease and TCM syndromes: The effect of clinical disease and TCM syndromes have statistically significant compared with control group(P <0.05).2.Comparison of TCM syndrome scores:The two groups of patients’ symptom scores showed no significant difference before treatment(P> 0.05); Treatment group treat for 4 weeks and 8 weeks compared with no treatment, the difference were statistically significant(P <0.01); the treatment group and control group treat for 8weeks scores declined compared with no treatment, the difference was statistically significant(P <0.01); treatment group treat for 4 weeks and 8 weeks scores declined better than the control group, the difference was statistically significant(P <0.01).3.Comparison of ACR and 24 h UPr:There was no significant difference in ACR and 24 h UPr before treatment(P> 0.05); In ACR and 24 h UPr,treatment group treat for 4weeks and 8 weeks were more significant difference compared with no treatment(P<0.01); In ACR and 24 h UPr,control group treat for 4 weeks and 8 weeks were more significant difference compared with no treatment(P <0.01), treat for 8 weeks compared with 4 weeks were more significant difference(P <0.05);In ACR and 24 h UPr,treatment group treat for 4 weeks compared with the control group, there was a significant difference(P <0.05), treatment of 8 weeks compared with the control group,there was a very significant difference(P <0.01).4.Comparison of renal function(BUN、Scr) and glucose(FPG、2h PG、Hb A1C) :The two groups of patients with no treatment and 4 weeks treatment and 8 weeks treatment were no significant difference in renal function and blood glucose(P> 0.05);Renal function and blood glucose have no significant difference between the treatment group and control group(P> 0.05).Conclusions:Tang-Shen-Kang can improve the clinical symptoms,reduce urinary protein and protect kidney. |
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