Based On Oxidative Stress, The Mechanism Of Yangyin Yiqi Huoxue Method For Prevention And Treatment Of Macrovascular Disease In GK Rats Was Studied

Objective:The study is to explore the molecular mechanism of action that nourishing Yin benefiting Qi and invigorating the circulation of blood therapy prevents and cures large vascular disease of diabetes mellitus (DM) in the aspects of differential gene expression and correlative metabolic pathways, to provide the scienti-fic basis for traditional Chinese medicine's prophylaxis and treatment on this disease starting with oxidative stress and applying the method of the whole genome expression profiling to seek the molecular mechanism of nourishing Yin benefiting Qi and invigorating the circulation of blood therapy for preventing and curing this disease.Methods:Randomly divide 160 7-8 week-old male rats, fasting blood glucose ? 11.0mmol/L, with spontaneous T2DM into 5 groups, Model Group. ShenQi Compound High Dose Group, ShenQi Compound Middle Dose Group, ShenQi Compound Low Dose Group and Metformin Group. Continuously feed these 5 groups with high fat diet for 12 weeks to create the metabolic memory model of large vascular disease of DM. Group another 32 rats as Blank Group and feed with normal diet.After 16 weeks'intervention therapy, drench Model Group with hysiological saline 10g/kg.d/rat, ShenQi Compound High Dose Group with ShenQi compound extract 28.8ml/kg.d/rat, ShenQi Compound Middle Dose Group with ShenQi compound extract 14.4g/kg.d/rat. ShenQi Compound Low Dose Group with ShenQi compound extract 7.2g/kg.d/rat and Metformin Group with metformin 167g/kg.d/rat Oberve the drinking amount and food-intake status of GK rats during the experiment. At the week 0,8 and 16 during the intervention treatment, check the blood sugar, blood fat, blood ox-LDL and 3-NT of 10 GK rats per group randomly. Besides, check the aorta vascular tissue's SOD and GSH-Px's level and activity by cellular level,process morphological observation on aorta by HE and Masson staining of 6 GK rats per group randomly, compare the effects of oxidized stress reaction among different intervention treatment. In week 16, based on the differential genes acquired from expression profile chip technology, applying the GO Enrichment and pathway analysis, analyze the molecular mechanism affected by ShenQi compound by combining the gene expression profile.Results:The drinking amount and food intake of the ShenQi Compound groups were less than the Model Group.ShenQi Compound groups could effectively decrease the relevant index level of blood sugar, blood fat and oxidized stress reaction, decrease collagen fiber and vascular wall area ratios significantly.In week 8, ShenQi Compound Middle Dose Group could significantly improve the aorta vascular tissue's SOD, GSH-Px, blood 3-NT and ox-LDL's level and reduce the oxidized stress reaction caused by high glucose.It was observed with microscope that ShenQi Compound can protect vascular endothelial injury caused by high sugar, decrease collagen fiber,especially to High and Middle Dose Group.Compared ShenQi Compound Middle Dose Group with Model Group, differential expression genes and signaling pathway were Sod1, Sod2, Ponl, Pon3, Elavl1, Lama4, JAK/STAT signaling pathway and Notch signaling pathway. Analyzed by GO Enrichment and Pathway, it mainly related to positive regulation of angiogenesis?cell death?cell dhesion?cellular macromolecule metabolic process.Result of light-microscope observation indicates that ShenQi compound could prevent the harm on the vessel's eSODerma effectively, reduce the death of the cells of the vessel's eSODerma of the experimental rats,especially for the ShenQi Compound Middle Dose Group in week 8.Conclusion:Nourishing Yin benefiting Qi and invigorating the circulation of blood therapy achieved the best treatment effect on intervening the blood suger and fat's metabolic disturbance, oxidized stress reaction, harm on the vessel's eSODerma and arteriosclerosis which are produced by large vascular disease of DM in week 8. The mechanism of action might be that ShenQi Compound can reduce the harm of the oxidized stress reaction to the vessels, decrease collagen fiber, inhibits angiogenesis, and promote the recovery of the cells of the vesse's eSODerma at the early stage by effecting correlative genes and signaling pathways such as Sodl, Sod2, Ponl, Pon3? Elavl1 Lama4, JAK/STAT signaling pathway and Notch signaling pathway as well, to prevent and cure large vascular disease of DM.