On The Role And Mechanism Of The Intervention On Diabetic Rats Macroangiopathy By Inhibition Of NF-κB Signal Pathway By Astragaloside And Ferulic Acid

ObjectiveThe dynamic change of NF-κB pathway to observe the effective intervention ofdiabetic rat vascular endothelial cell structure,function and regulation factor based on,toinvestigate the protective effect and mechanism of ASTIVragaloside and ferulic acid invascular endothelial injury and dysfunction in diabetic rats.MethodsRandomly selected8week old SPF male Wistar rats60,weight200±20g.Wererandomly divided into control group (n=10) and model group(n=50).The STZ induceddiabetic rats were randomly divided into model group (group DM, n=10), Astragalosidegroup(ASTIV,50mg/kg,D,n=10),ferulic acid group (FA,50mg/kg,D,n=10),group ofastragaloside and ferulic acid (ASTIV/FA,dose ibid,n=10).Control group,metformin group(DMBG,40.5mg/kg,D,n=10).AST group,FA group and AST/FA group were dissolved in55℃saline,by using now,gastric gavage daily at9Am.Model group and blank group weregiven normal saline.Results1.Effectting on blood glucose、blood lipid：Compared with the rats in the blank group,model group and the treatment group blood sugar in all around, was rised after eight weeks(P<0.01)，DMBG significantly decreased the blood glucose of model group（P<0.01）,ASTgroup,AST/FA group was decreased than in model group（P<0.05）,The FA groupdecreased blood sugar was not obvious,Compared with the control group,TC and TG levelswere significantly elevated(P<0.01),Compared with the model group,GroupAST/FA,group FA and TG were significantly lower in TC（P<0.01）;Group DMBG did not affect TC and TG content（P>0.05）.Before and after the treatment,liver function,renalfunction showed no difference between groups.2.Comparison of the level of NO in the rats in the model group and normal groupdecreased,the drug group were lower than model group increased(NO P<0.05),whereASTIV/FA group increased most obviously,there was significant difference compared withmodel group(P<0.01).3.Light microscopic observation of pathologic changes of aortic tissue:The controlgroup of vascular wall structure clear,complete and smooth intima,endothelial cell integritywithout shedding,smooth muscle cell proliferation,without walking clear;The model groupthickened vessel wall endothelial cells,most of the missing,the proliferation of smoothmuscle cell was arranged in disorder;ASTIV/FA group vascular intimal smooth andcontinuous,endothelial cells occasionally fall off,the proliferation of smooth muscle cell isnot obvious,uniform thickness,regular shape,no obvious abnormality.4.Detection of ELISA expression in model group,serum oxLDL increasedobviously,there was significant difference compared with the control group(P<0.05).Group FA,group ASTIV/FA significantly inhibited the expression of oxLDL indiabetic rats,compared with the model group were statistically significant(P<0.01).5.To detect the expression of ELISA in aortic tissue of rats in model groupMCP-1,TNF-αincreased obviously,there was significant difference compared with thecontrol group (P<0.05).Group FA,group DMBG had certain inhibition on diabetic ratsMCP-1,TNF-α,Group ASTIV,group ASTIV/FA significantly inhibited MCP-1,TNF-αdiabetic rats expression.6.Immunohistochemistry compared with the control group,model group ratsabdominal aorta endothelial area MCP-1,TNF-α, NF-κB protein positive cellsnumbe.Shading significantly enhanced,The number of ASTIV/FA group in rat aorticendothelial cells at least,tinting strength is shallow;Group ASTIV,group FA expressionthan western medicine control the number of positive cells were less than ASTIV/FAgroup,but the number of positive staining;western medicine control group compared withmodel group,the positive expression.7.WesterBlot detection showed that in model group rats abdominal aorta endotheliumphosphorylation of NF-κBp65increased significantl;Compared with model group,eachgroup of phosphorylation of NF-κBp65decreased,the difference was statisticallysignificant(P<0.05). 8.RT-PCR detection showed that the expression of MCP-1、TNF-α was significantlyhigher in the model group,Inhibitory effect of each treatment groups were expressed inendothelial tissue MCP-1mRNA、TNF-αmRNA significantly,And the results were thesame, Compared with the model group were significantly different(P<0.05);Expression ofASTIV/FA group protein compared with traditional Chinese medicine monomergroup,there were significant differences(P<0.05).Conclusion1.The aorta in diabetic rats phosphorylated NF-κBp65expression induced bySTZ,suggesting that B may be involved in the pathogenesis of NF-κB macrovasculardiabete.2.Elevated oxLDL can enhance the NF-κBp65activity of the protein,which lead to thehigh expression of NF-κB pathway downstream inflammatory factor MCP-1,TNF-α in theserum and aorta in diabetic rats.3.Expression of astragaloside ferulic acid can reduce diabetic rat aortic NF-κBp65activation, in addition to inhibit NF-κB pathway through the activation of blood glucosecontrol,but also to inhibit NF-κB pathway by reducing oxLDL,TNF-α and othermechanism. And they have synergistic effect.