| Objective:Explore the effection of tang-shen-kang on the kidney tissues' ICAM-1 and MCP-1 of diabetic nephropathy(DN) rats with symptoms of qi-yin defi-cuenca and blood stasis syndrome,and confirmed the mechanism of action of Tang-shen-kang on the curative effect of DN rats.Methods:Divided the 50 healthy male rats into normal group 8 and model 42 by simple random sampling method. Then successful model rats were randomly divided into 4groups: Model group, Tang-shen-kang group, Benazepril group, Antioxidants pyrrolidine dithiocarbamate(PDTC) group. Before the experiment blood glucose,urinary albumin / creatinine ratio(ACR) were measured in each group of rats,then the negative results for experimental study. In addition to the normal group of 8 rats, 42 rats in model group were used by triple law(surgery + high-sugar and high-fat diet +streptozotocin)(STZ) to make the DN animal model. After making the model successfully, normal group and model group fixed with animal feed and tap water feeding. Tang-Shen-Kang group were gavaged with 1.6g/(200g· d)Tang-Shen-Kang dissolved in warm water. Benazepril group were gavaged with 0.3mg/( 200g·d)benazepril dissolved in warm water. PDTC group with PDTC dissolved in saline to8mg/(200g·d) by intraperitoneal injection. The rats were gavaged or intraperitoneal injection once a day and 8 weeks of treatment. After the end of the experiment each group rats abdominaled aorta blood, and removal of the right kidney by surgical operation. After quickly weighed,the kidney were kept in neutral formalin solution.All the rats urined specimens before surgery. Detecte BUN by enzyme-coupled rate method.detecte Scr by creatinine enzymatic. detecte ACR by radioimmunoassay.detecte MCP-1,ICAM-1 in tissues of rats by immunohistochemistry.detect the levels of MCP-1, ICAM-1 by ELISA. observed pathological changes in renal tissues by HE and Masson+PASM.Results:1. The change of TCM symptoms and signs in rats before and after treatment:The normal group's food intake and water conditions, mental status, daily activities were normal, weight steady increased in rats. Model group is manifested polydipsia, polyphagia, polyuria, gradual weight loss, listlessness, activity slow, and the symptoms gradually worsened with the extension of time. The Tang-shen-kang group's food intake and water conditions gradually increased, mental state is significantly improved,compared with model group, the hair shiny and activity gradually increased, weight stable. Benazepril group and Tang-shen-kang group have similar performance. PDTC group between model group and Tang-shen-kang group, Benazepril group. The rats in the normal group have no death.6 rats was died during making models, the cause may be the anesthetic overdose, side effects after injection STZ, surgical infections. Model group and PDTC group have died 3 rats after treatment, the cause of death may be biting each other, malnutrition, improper gavage and so on. Tang-Shen-Kang group and benazepril group have died 1 rat after treatment.After the death we found severe gastrointestinal obstruction by dissection.2. Comparison of body weight and kidney hypertrophy index of rats in each group:Body weight, by One-way ANOVA,compared with normal group of the same period,the model group, Tang-shen-kang group, benazepril group and PDTC group rats' weight were reduced(P<0.05 or P<0.01). The end of 2 weeks and 4 weeks of treatment,Tang-shen-kang group, benazepril group and PDTC group have no significant. The end of the 6 weeks and 8weeks, compared with model group, Tang-shen-kang group,benazepril group, and PDTC group were increased,it has significant difference(P<0.01).Tang-shen-kang group and benazepril group have no significant(P>0.05). PDTC group's weight was significantly reduced compared with Tang-shen-kang group and benazepril group(P<0.05 or P<0.01). Kidney hypertrophy index, by One-way ANOVA,compared with normal group of the same period, the model group, Tang-shen-kang group,benazepril group and PDTC group rats' weight were significantly increased(P<0.01).Tang-shen-kang group and benazepril group and PDTC group were significantly reduced(P<0.01). Tang-shen-kang group and benazepril group have no significant(P>0.05).PDTC group was significantly increased when it compared with Tang-shen-kang group and benazepril group(P<0.01).3. Comparison of each groups' renal function(BUN?Scr): By one-way ANOVA, the rats' BUN and Scr have no significant difference(P> 0.05).4. Comparison of each groups' ACR: By one-way ANOVA, compared with normal group, the model group, Tang-shen-kang group, benazepril group and PDTC group rats' ACR were significantly increased(P<0.01). The 2 weeks and 4 weeks of treatment,Tang-shen-kang group,benazepril group and PDTC group were significantly reduced when they compared with model group(P<0.01). Tang-shen-kang group and benazepril group have no significant(P>0.05). PDTC group was significantly increased when it compared with Tang-shen-kang group and benazepril group(P<0.01). At the end of 4weeks Tang-shen-kang group,benazepril group and PDTC group showed no significant difference with the treatment of the end of 8 weeks(P>0.05),but have significant difference compared with no treatment(P< 0.05 or P <0.01).5. Comparison of each groups' expression of MCP-1, ICAM-1: By one-way ANOVA, compared with normal group, the model group, Tang-shen-kang group,benazepril group and PDTC group rats' MCP-1, ICAM-1 were significantly increased(P<0.01). Tang-shen-kang group,benazepril group and PDTC group were significantly reduced when they compared with model group(P<0.01). Tang-shen-kang group and benazepril group have no significant(P>0.05). PDTC group was significantly increased when it compared with Tang-shen-kang group and benazepril group(P<0.01).6. The correlation analysis between ACR and MCP-1, ICAM-1: The rats' ACR and MCP-1, ICAM-1 were positively correlated(r>0,P<0.01).7. Renal pathological morphology:(1)The naked eye observation of renal tissue:the size of normal group in the the right kidney was normal, the color is dark red. While the model group rats' volume of right kidney increased obviously. Compared with normal group, Tang-shen-kang group and benazepril group were increased,but smaller than the model group. PDTC group between model group and Tang-shen-kang group,benazepril.(2)HE staining: Glomerular size and shape of the normal group were normal.The renal capsule cavity of model group was significantly reduced. Tang-shen-kang group and benazepril group slightly smaller. PDTC groups' wsa between model group and Tang-shen-kang group, benazepril. All the five groups' Mesangial cells and endothelial cells showed no significantly increased.(3)Masson+PASM staining:The performance of glomerular basement membrane(GBM): Compared with the normal group, the model group was significantly thicker, this is the basic pathological changes in DN. Benazepril group showed no significant difference Tang-shen-kang group,compared with the normal group slightly thickened, but significantly improved compared with the model group. PDTC groups' was between model group and Tang-shen-kang group, benazepril group. The performance of mesangial matrix: Model group was significantly increased compared with the normal group. Compared with the normal group,Tang-shen-kang group and benazepril group showed slightly increased,but decreased significantly compared with model group. And Tang-shen-kang group and benazepril group have no significantly. PDTC group showed increased than Tangshenkang group and benazepril group, but reduced compared with the model group.Conclusions:1. Tang-Shen-Kang can improve the quality of life of DN animal models, and reduce urinary protein of DN rats.2. Tang-Shen-Kang can improve the renal morbid pathology state of DN animal models.3. The DN animal models' expression of MCP-1, ICAM-1 were significantly increased,this means that NF-?B signaling channel activator involved in the pathogenesis DN animal models. And significantly positively related to ACR. This suggests that the inflammatory cytokines may be involved in the occurrence and development of DNObjective:By observing the clinical efficacy of Tang-Shen-Kang on the DN of qi-yin defi-cuenca and blood stasis syndrome patients and the changes of MCP-1,ICAM-1 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 defi-cuenca and blood stasis syndrome.Methods:Select 66 patients of qi-yin defi-cuenca and blood stasis syndrome admitted in our department and endocrinology from May 2014 to January 2016,then divided into treatment group and control group 33 cases. In the course of treatment we lost 3 cases,including 1 cases of treatment group, 2 cases of the control group, 63 cases completed.The other 30 healthy people were selected as the normal group. The two groups of patients were treated with comprehensive treatment to control diet, blood glucose and blood pressure.The treatment group took mixed Tang-Shen-Kang on the comprehensive treatment of the above, 3 times a day, each time a bag, for 8 weeks.We observed two groups with ACR, MCP-1, ICAM-1, renal function, blood glucose and other indicators and the occurrence of adverse reactions.The nomal group observed the MCP-1, ICAM-1 only.then took statistical analysis and determine the efficacy according to the experiment results.Results:1. Comparison of the clinical efficacy of disease: The total effective rate of treatment group and control group were 90.63%,70.97%.By rank sum test, P<0.05.This means that the treatment group was better.2.Comparison of ACR: By t-test,before treatment the two groups of patients have no significant difference(P>0.05). After treatment The ACR of treatment group and control group were significantly reduced(P<0.01).And the treatment group was better(P<0.01).3. Comparison of MCP-1,ICAM-1: By t-test,before treatment, compared with the normal group, the levels of treatment group and the control group were significantly increased,(P <0.01); Treatment group and control group had no significant difference(P> 0.05).After treatment, the levels of treatment group and control group were significantly reduced, the difference was statistically significant(P <0.01). The decrease compared with the treatment group and the control group was statistically significant(P<0.05).4. Comparison of renal function(BUN?Scr) and glucose(FPG?2h PG?Hb A1C) :Before treatment the two groups of patients have no significant difference(P>0.05).The treatment group and control group have no significant difference after treatment(P>0.05).Conclusions : Tang-shen-kang can significant reduced the MCP-1,ICAM-1,urinary protein of qi-yin defi-cuenca and blood stasis syndrome patients and Tang-shen-kang can improve the clinical symptoms, protect kidney. |
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