| Objective:To observe the express and variance of Tangshenkang on rats in the blood of NF-?B p65?p-I?B??IKK?.To investigate the function mechanism of Tangshenkang on DN Rats.Methods:Totally 40 healthy male SD rats were randomly divided into normal group(8cases) and model group(42 cases) by randomized block method,then successful model rats were randomly divided into 4 group:Model group,Tang-Shen-Kang group,Benazepril group.Rats in each group pre-test blood glucose,24 hours Urine Protein(24h UPr)were 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 with1.6g/(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.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; To detect the expression of renal tissue with immunohistochemical method in NF-?B p65?p-I?B??IKK?.Detecting changes in NF-?B p65, p-I?B?, IKK? in the blood of in rats by ELISA.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.The performance of model rats as polydipsia, polyphagia, polyuria,gradual weight loss, listlessness, activity slow, unresponsive, coat haggard and so easy to take off.With time, the symptoms of rats gradually worsened.Food intake and water intake in Tang-Shen-Kang group gradually decreased, 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.Normal rats no deaths. triple modeling method died five,considered due to anesthetic overdose, side effects after the injection of STZ, surgical infections; model group died three during the experiment, the cause of death rat bite each other, due to improper oral; Tangshenkang group and benazepril group died each one, autopsy found serious rat gastrointestinal tract obstruction after the death.2.Comparison of body weight of rats in each group and kidney hypertrophy index:By one way ANOVA, Compared with the same period in normal group, the rest weight of rats between each group were significantly reduced(P<0.05 or P<0.01).In 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).In the 6 weeks and 8 weeks of treatment, Tang-Shen-Kang group and benazepril group compared with model group have significant difference(P<0.01), Tang-Shen-Kang group have no significant difference compared with benazepril group(P>0.05). In 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 with each group(BUN?Scr):By one way ANOVA, the rats BUN and Scr have no significant difference(P>0.05).4.ACR compares in each group:By one way ANOVA, compared with normal group,ACR in each group increased significantly(P<0.01). compared with model group,Tang-Shen-Kang group and benazepril group were decreased significantly in 4 weeks and 8 weeks of treatment(P<0.01),Tang Shen Kang group have no significant difference with benazepril group(P>0.05).8 weeks of treatment compared with 4 weeks of treatment have no significant difference in Tang-Shen-Kang group and benazepril group(P>0.05),but have significant difference compared with untreated(P< 0.05 or P <0.01).5.The expression of NF-?B p65 in rat comparison(ELISA method): By one way ANOVA, Compared with normal group, NF-?B p65 of PBMC levels in each group were higher than the normal group, the difference was significant(P<0.01); compared with the model group, Tangshenkang group and NF-?B p65 in PBMC levels benazepril group were on average lower than the model group, the difference was significant(P<0.01); Tangshenkang group and shellfish Nafplio Lee rats in PBMC NF-?B p65 levels showed no significant difference(P> 0.05).6.The expression of p-I?B? in rat comparison(ELISA method): By one way ANOVA,Compared with the normal group, the rats in the p-I?B? PBMC were lower than the normal group, the difference was statistically significant(P<0.05); compared with the model group, Tangshenkang group and benazepril group PBMC in p-I?B? levels were higher than the model group, the difference was statistically significant(P<0.05); large Tangshenkang group and benazepril group murine serum levels of p-I?B? was no significant difference(P>0.05).7.The expression of IKK? in rat comparison(ELISA method): By one way ANOVA,Compared with the normal group, the rats in PBMC IKK? levels were higher than the normal group, the difference was significant(P<0.01);compared with the model group,Tangshenkang group and benazepril water rat serum IKK? Plymouth group, the average in the model group, the difference was significant(P<0.01); Tangshenkang group and benazepril IKK? serum levels in rats Plymouth group was not significant difference(P>0.05).8. The correlation analysis of the rats ACR and of NF-?B p65, p-I?B?,IKK? :ACR in rats and NF-?B p65, IKK? in blood were positively correlated(r>0, P <0.01); ACR in rats and p-I?B? in blood was negatively correlated(r<0, P<0.01).9.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;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 renal capsule was significantly narrow in model group and disease- syndromeme model group.The renal capsule narrow 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; 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. Benazepril group and Tang-Shen-Kang group roughly equal, compared with the normal group increased slightly, but significantly reduced compared with the model group.Conclusion: 1.NF-?B activation is involved in the pathogenesis of the signal path DN rat model;2. Tangshenkang capable of inhibiting NF-?B p65, IKK?, improve the level of p-I?B?,which may be one of the mechanisms of the efficacy of treatment Tangshenkang DN;3. Tangshenkang can improve the quality of life of DN model rats, reduce urinary protein DN rats.Objective: By observing the changes of the Tangshenkang on the DN Qi-Yin deficiency and blood stasis syndrome patients of ACR and the blood of NF-?B p65?p-I?B??IKK?before and after treatment,to explore the renal protective effect of Tangshenkang on the DN Qi-Yin deficiency and blood stasis syndrome patients.Methods:66 cases DN patients with Qi-Yin deficiency and blood stasis syndrome from May 2014 to January 2016 in Department of Nephrology and Endocrinology were randomly divided into treatment group(33 cases) and control group(34 cases).In the course of treatment lost 3 cases, two cases in the treatment group and one cases in the control group, the actual completion of 63 cases, and 20 cases of normal healthy persons as normal group.The two groups of patients were given comprehensive treatment to control diet, blood pressure and blood glucose; 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.Observing the efficacy of disease, ACR,kidney function,blood glucose, etc.Observing the adverse reactions and the level of NF-?B p65?p-I?B??IKK?in blood,only the normal group detection blood of NF-?B p65?p-I?B??IKK?.According to the experimental results,the statistically will be analyzed and the efficacy will be determined.Results:1. Comparison of efficacy of clinical disease : disease treatment group total effective rate was 90.63%,70.97% in the control group, there were significant differences between the two groups(P <0.01).2. Comparison of ACR: There was no significant difference in ACR before treatment(P>0.05);There was significant difference in the treatment group ACR before and after treat(P<0.01);There was significant difference in the control group ACR before and after treat(P<0.01); In ACR,the treatment group after treat compared with the control group,there was a significant difference(P<0.01).3.Comparison of NF-?B p65 ? p-I?B? ? IKK? : NF-?Bp65 ? IKK? were significantly higher than normal group before treatment between two groups of patients(P<0.01), p-I?B? was significantly lower than normal group(P<0.01), while the comparison between each treatment group and the control group were no significant difference(P>0.05);After treatment, the NF-?B p65,IKK? levels than before treatment were significantly decreased(P<0.05), p-I?B? levels than before treatment were significantly increased(P<0.05),and treatment level group improved significantly better than the control group(P<0.05).4.Comparison of renal function(BUN?Scr)and glucose(FPG?2h PG?Hb Al C): The two groups of patients with untreated 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: 1.Tangshenkang can significantly improve the clinical symptoms of DN Qi-Yin deficiency and blood stasis syndrome, reduce urinary protein,with good clinical efficacy. 2. Tangshenkang can reduce the levels of NF-?B p65?IKK?,raise the level of p-I?B? in DN Qi-Yin deficiency and blood stasis syndrome.Indicating Tangshenkang can inhibit NF-?B signaling pathway, which may be one mechanism of clinical efficacy DN Tangshenkang particle therapy.3.In the treatment of patients with damp heat syndrome after the detection of safetyindicators showed no abnormal changes,the phenomenon of adverse reactions drug did not appear in the process,good safety instructions Tangshenkang. |
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