Study On The Distribution Characteristics Of Syndrome Factors Of Diabetic Nephropathy And The Mechanism Of "From The Wind Treatment" Based On PI3K / Akt / NF-κB Pathway

Section I Clinical Research Research on syndromes characteristics of diabetic kidney disease based factorial analysisObjectivesTo explore the pathogenesis of this disease and provide evidence for diagnosis and treatment of diabetic kidney disease by Chinese Traditional Medicine, research on syndromes distribution characteristics of diabetic kidney disease in early and clinical stage.MethodsFour diagnostic information was collected based on cross-sectional survey of patients with diabetic kidney disease in this clinical study. Similar symptoms were organized by factorial analysis,useing the professor Wenfeng Zhu a book about syndromes characteristics and expert assessment with choosing the most suitable factors.Calculating the factors score of each patient according to factorial analysis was to identify different types of each patient.ResultsTwelve factors were extracted in diabetic kidney disease of early stage that include Qi deficiency,Xue deficiency,Yang deficiency,Yin deficiency,wind pathogen,phlegm-damp and blood stasis.The early location of DKD was in spleen,kidney, liver, stomach and lung.The incidence rates of total symptoms were ranked from high to low as follows:Qi deficiency 84.8%,Yin deficiency 70.9%,phlegm 69.6%,Xue deficiency 67.7%,Yang deficiency 57%, blood stasis 55.7%,damp 41.8%and wind pathogen 39.9%. The incidence rates of dominating symptoms were ranked from high to low as follows:Qi deficiency28.5%,Yang deficiency 19.6%,dampl5.2%,Yin deficiency 12.7%,blood stasis7%,phlegm7%,wind pathogen6.3% and Xue deficiency6%.Eleven factors were extracted in diabetic kidney disease of clinical stage that include Qi deficiency,Xue deficiency,Yang deficiency,Yin deficiency,wind pathogen, phlegm-damp and blood stasis.The clinical location of DKD was in heart, spleen, kidney, liver, stomach and lung.The incidences rate of total symptoms were ranked from high to low as follow:Qi deficiency74.5%,damp70.8%,blood stasis64.6%,Yin deficiency62.1%,wind 44.7%,Yang deficiency42.9%, phlegm39.1% and Xue deficiency 33.5%.The incidence rates of dominating symptoms were ranked from high to low as follows:Yang deficiency 18%, wind pathogen 24%,Qi deficiency 14.3%,damp 13.0%,Xue deficiency 11.8%, Yin deficiency 10.6%,blood stasis 9.9%,phlegm7%.ConclusionThe features of diabetic kidney disease are deficiency and excess in early and clinical stage.The deficiency include Xue deficiency,Yang deficiency,Yin deficiency and Qi deficiency especially.The excess include wind pathogen,particularly phlegm-damp and blood stasis.We should pay attention to dispel wind and remove obstruction in the meridians on the basis of supplementing qi and activating blood circulation in view of the pathogenesis of diabetic kidney disease.Section II Experimental Research Research on treatment of diabetic kidney disease from the wind governs based PI3K/Akt/NF-KB signaling pathwayObjectivesDiabetic nephropathy present kidney damage caused by diabetes, called diabetic kidney disease in clinical diagnosis and diabetic glomerulosclerosis in pathological diagnosis.We identified the mRNA and protein expression of p-PI3K,p-Akt,PTEN,NF-KB and MCP-1 to explore the mechanism of the granule of YiQiSouFengTongLuo based PI3K/Akt/NF-κB signaling pathway mediated immune inflammatory response,which provided the experimental basic data for the treatment from the wind governs.MethodsThe GK rat model of diabetic kidney disease was established by uninephrectomy and feeding on high caloric and fat diet. The GK rat was randomly assigned model group, sham-operated group,Yiqi group,Qufeng tongluo group and Yiqi soufeng tongluo group.All the rats were intragastric administration for 8 weeks and collected urine samples over 24 hours respectively in the 4th,6th and 8th week for an accurate measure of the protein in their urine with evaluating therapeutic effects and prognosis. At the 8th weeks, rats were sacrificed to measure GLU,Scr,ALT,AST and TBIL.We identified the mRNA and protein expression of p-PI3K, p-Akt, PTEN,NF-κB and MCP-1 and observed the pathological section with HE stain method to explore the mechanism of TCM.ResultsIn experimental study 1,Urine protein quantitation in 24 hours of each group of rats was no statistically differences in the early experimental stage.Compared with model group urine protein quantitation in 24 hours, the other groups decline in the 4th week. Yiqi group and Yiqi soufeng tongluo group had statistically significant difference(P< 0.01). Qufeng tongluo grouphad statistically difference(P< 0.05).there was no statistic significance in else. Compared with Yiqi soufeng tongluo,Yiqi group decline and Qufeng tongluo group elevate.But,they were no statistic significance. Compared with model group,the other groups declined in the 6th week. Yiqi group had statistically significant difference(P<0.01).Qufeng tongluo group and Yiqi soufeng tongluo group had statistically difference(P<0.05).there was no statistic significance in else.Compared with Yiqi soufeng tongluo group,Yiqi group declined.But,it was no statistic significance.Compared with model group,the other groups declined in the 8th week. Yiqi group had statistically significant difference(P< 0.05). Yiqi soufeng tongluo group had markedly statistically significant difference(P<0.05).there was no statistic significance in else.Compared with Yiqi soufeng tongluo group,Yiqi group,Qufeng tongluo group elevated.Qufeng tongluo group has statistically difference(P<0.05).Compared with model group Scr,the other groups declined, sham group had statistically markedly significant difference(P< 0.01).there was no statistic significance in else.The glucose,ALT and AST of each group was no statistically differences.In experimental study 2,we detectd the expression of p-PI3K,p-Akt,PTEN,NF-KB and MCP-1 by immunohistochemical method.The result showed that Compared with the p-PI3K expression of model group,the other groups reduce.Yiqi soufeng tongluo group had statistically significant difference(P< 0.01).there was no statistic significance in else. Compared with Yiqi soufeng tongluo group,Yiqi group reduced and Qufeng tongluo group increased.But,there was no statistic significance.Compared with the p-Akt expression of model group,the other groups reduce. Yiqi soufeng tongluo group had statistically significant difference(0.05).there was no statistic significance in else.Compared with Yiqi soufeng tongluo group,Yiqi group and Qufeng tongluo group increased.But,there was no statistic significance. Compared with the expression of PTEN model group,sham group,Yiqi group,Qufeng tongluo group and Yiqi soufeng tongluo group increased. But,there was no statistic significance.Compared with Yiqi soufeng tongluo group,Yiqi group and Qufeng tongluo group reduced.But,there was no statistic significance.There was no differences in the expressione of NF-κB and MCP-1 among each groups, we detectd the protein and mRNA of p-PI3K,p-Akt,PTEN, NF-κB and MCP-1 expression in the kidney tissue measured by western blot and real-time PCR respectively. Compared with the protein expression of p-PI3K model group,the other groups reduced. They all have statistically significant difference except sham group(P<0.01).Compared with Yiqi soufeng tongluo group,Yiqi group increased and Qufeng tongluo group reduced.But there was no statistic significance. Compared with the expression of p-Akt model group,treatment group reduced. Yiqi group and Qufeng tongluo group had statistically significant difference(P< 0.05).Yiqi soufeng tongluo group had statistically significant difference(P<0.01).there was no statistic significance in else.Compared with Yiqi soufeng tongluo group,Yiqi group and Qufeng tongluo group increased.But,there was no statistic significance.There was no differences in the expressione of PTEN among each groups. Com-pared with the NF-κB expression of model group,the other groups reduced. Yiqi group, Qufeng tongluo group and Yiqi soufeng tongluo group had statistically significant difference (P< 0.01).there was no statistic significance in else.Compared with Yiqi soufeng tongluo group,Yiqi group reduced and Qufeng tongluo group increased.Yiqi group and Qufeng tongluo group had statistically significant difference(P< 0.01).Compared with the MCP-1 expression of model group,treatment group reduced. Yiqi group and Yiqi soufeng tongluo group had statistically significant difference(P<0.01).Qufeng tongluo group has statistically difference(P< 0.05). Compared with Yiqi soufeng tongluo group,Yiqi group reduce and Qufeng tongluo group increase.But,there was no statistic significance.Compared with the mRNA expression of PI3K model group,the other groups reduced. The treatment had statistically significant difference (P< 0.01).Compared with the mRNA expression of Akt model group,sham group increase and the other groups reduced. They all had statistically significant difference.Compared with the mRNA expression of PTEN model group,the other groups reduced. The treatment had statistically significant difference (P< 0.01).Compared with sham group,NF-κB mRNA and MCP-lmRNA expression of the other groups reduced. They all had statistically significant difference.Compared with Yiqi soufeng PI3K mRNA,Akt mRNA,PTEN mRNA,NF-KB mRNA and MCP-lmRNA expression of tongluo group,the others increased.Yiqi group had statistically significant difference (P< 0.01),there was no statistic significance in else.Conclusions1 The GK rat model of diabetic kidney disease that is established by uninephrectomy and feeding on high caloric and fat diet is a relatively ideal mode.Its advantage is that simulating the pathogenesis of DKD from a very large extent with using Spontaneous model of type 2 diabetes.2 Compared to Astragalus or Qufeng tongluo granule individually, Yiqi soufeng tongluo granule can reduce urine protein quantitation in 24 hours. Results show that the efficacy is better with the advance testing time.3 Yiqi soufeng tongluo granule can protect renal function via decreasing the protein and mRNA expression of p-PI3K,p-Akt,NF-KB and MCP-1 to regulate the PI3K/Akt/NF-κB signaling pathway.The effect is better than Astragalus or Qufeng tongluo granule individually.