Law Of Diabetic Peripheral Neuropathy Certificate Candidates And Sugar Bi Kang Intervention High Sugar Environment Endothelial Cells Mechanism Studies

Objective:Diabetic peripheral neuropathy is a common complication of diabetes.My Ph.D. advisor Professor Liu Tonghua has accumulated rich experience in this respect.He holds the opinion that "poison" and "stasis" play a important role in the pathogenesis of diabetic peripheral neuropathy.The paper has two parts:clinical research and experimental research.In the clinical research part,we collect cases of diabetic peripheral neuropathy patients to establish a database of diabetic peripheral neuropathy and use the K-means cluster analysis to analyze syndromes of diabetic peripheral neuropathy.In the experimental research,we study the mechanism of Tang Bi Kang s intervening in human umbilical vein endothelial cells which are damaged by high-sugar envionment.We observe the function of rat serum containing Tang Bi Kang to human umbilical vein endothelial cells which is curtured in high sugar environment.Tang Bi Kang is the candidates drug of diabetic peripheral neuropathy.This research study the treatment mechanisms of diabetic peripheral neuropathy on cell and molecular level from the perspective of the microvascular injury.We hope that the research can provide gist for the clinical application and development of new drugs.Method:Clinical research:200 patients who were diagnosed as diabetic peripheral neuropathy were collected,aged between 18-79 years,treatmented in the Dongfang Hospital beteen 2010-2011.Then filled the symptom information to establish a database of DPN. Further,filtered the data of symptoms,syndromes were analyzed with cluster analysis methods. At last, we concluded the result of the syndromes combined with clinic knowledge.Experimental research:Human umbilical vein endothelial cells were cultured in high-sugar environment,used CCK-8 method to assay the cell viability of different glucose concentration on human umbilical vein endothelial cells.Then,we assessd different glucose concentration on cultured HUVEC to supernatant NO.After modeling,studied Tang Bi Kang from the prospect of oxidative stress,protective effects of endothelial cells and peripheral nerves.There were three experiments:First,the influence of Tang Bi Kang on HUVEC s secreting LDH,MAD,SOD,NO who are in the high glucose environment. Second,the influence of Tang Bi Kang on the expression of VEGF,bcl-x,bax and eNOS mRNA under high glucose environment.Third,the influence of Tang Bi Kang on PI3K-Akt signaling pathway of HUVEC in the high glucose environment.Results:The clinical research:After filtering 123 TCM symptoms,ultimately,we selected 46 symptoms to analyze.After using a SPSS 17.0 software,K-means cluster analysis,combined with some expert advice,we divided DPN into 3 types:Yin and Yang deficiency, "poison" and "stasis" in neuron;Qi and Yin deficiency,"poison" and "stasis" in neuron;Qi deficiency, "poison" and "stasis" in neuron. Experimental part:Aftering comparing the conclusion of both endothelial cell growth activity and secreting NO of human umbilical vein endothelial cells under different concentrations of glucose,we concluded that the best environment for mode building of human umbilical vein endothelial cells were 25mmol/L glucose concentration.1,MDA and LDH level of HUVEC which were cultured under high glucose environment were higher than the normal group,but the NO content and SOD activity were lower compared with the normal group.Tang Bi Kang could significantly reduce the leakage of MDA and LDH of HUVEC which were damaged by high-sugar.Also,Tang Bi Kang could increase NO levels and SOD activity,compared with the normal group with statistical significance(P<0.05).2,Tang Bi Kang could increase VEGF gene expression in the high,middle and low dose groups(P<0.01).BaxmRNA were not statistically significant difference in different groups. Compared with the model group of high-sugar,Tang Bi Kang could significantly increased expression of bcl-xmRNA expression,furthermore Tang Bi Kang group were better than the Methycobal group(P<0.01).Compared with the model group of high-sugar environment,the high dose Tang Bi Kang group could significantly improve the eNOSmRNA expression and were better than the methylcobalamin group(P<0.05).High-sugar model,both the p85 and Akt phosphorylation levels were significantly decreased(P<0.01).Conclusion:The clinical research part:The K-means cluster analysis syndromes of DPN showed that:combining the retrospective survey and mathematical statistics method to study diabetic peripheral neuropathy syndrome type distribution of TCM were feasible.The results of this study can show the type of clinical DPN patient to a certain extent.Because the patient of this study were limited and from the same unit,so the results has relative limitations.The experimental part:Tang Bi Kang may improve the VEGFmRNA expression of endothelial cells,which play a role in vascular repair and protection.Through this,it can further improve neurovascular blood flow.Tang Bi Kang can improve the bcl-xmRNA,eNOSmRNA expression so as to inhibit apoptosis of endothelial cells and restore vessel endothelial function.Through this,it can play a protective effect on the surrounding nerve and blood vessels.AKT phosphorylation level of the Tang Bi Kang group is higher than the high-sugar level model group.Tang Bi Kang may protect endothelial cells through the phosphorylation signaling pathway.