Based On NF-κB And TGF-β1/Smads Pathway To Study The Mechanism Of Icariside Ⅱ Inhibiting The Carotid Intimal Hyperplasia After Balloon Injury Of Rats

Objective: To study the effect of icariin Ⅱ(Icariside Ⅱ,ICS Ⅱ)on intimal hyperplasia induced by balloon injury of carotid artery in rats and its possible mechanisms on p-NF-κB(p65),TGF-β1/Smads signaling pathway.Methods: SD rats(male)were randomly divided into sham operation group and the carotid artery intimal hyperplasia group,which was established by balloon injury method.After balloon injury,all balloon injury rats were randomly divided into model group,low dose and high dose group.ICS Ⅱ-L group and ICS Ⅱ-H group were given ICS Ⅱ(10,20mg/kg/d,qd,ig)for 14 d after balloon injury.The sham operation group and the model group were given distilled water as for the same volume for 14 d.The animals were disposed of 24 hours after the last dose.The pathological changes of blood vessels were observed by HE staining,Neointimal area(NIA)and the intimal area(IA)and media area(MA)was measured by Leica optical microscope in order to observe the changes of the neointimal hyperplasia.The expression of phosphorylated nuclear factor-κB(p-NF-κB)(p65)and IL-6 protein in carotid artery wall was detected by immunohistochemistry method and Western blot method.The expression of MMP-2,IκB-α,pSmad2,TGF-β1 and p-Smad3 as well as TNF-α in carotid artery wall were detected by Western blot method in all animals group.Results: Compared with the sham-operated group,the carotid intima proliferation of the injured side of the model group was significantly increased,the NIA was increased 11.2times(P<0.05),the ratio of IA/MA was increased 3.5 times(P<0.05),the positive expression rate of p-NF-κB(p65)was increased 2.9 times,and the expression of IL-6 was increased 2.0 times(P<0.05)detected by immunohistochemistry method).In addition,it was found that the protein expression of p-NF-κB,TGF-β1,p-Smad2,p-Smad3,MMP-2,IL-6,TNF-α were significantly increased(P<0.05),while the expression of IκB-α protein was significantly decreased(P<0.05)by Western blot method.Compared with Model group,carotid intima proliferation and the ratio of IA/MA in ICS Ⅱ-L group and ICS Ⅱ-Hgroup were significantly lower(P<0.05).The positive expression rate of p-NF-κB(p65)and IL-6 were significantly decreased by immunohistochemical method(P<0.05).Western blot analysis showed that the protein expression of p-NF-κB(p65),IL6,TNF-α,TGF-β1,p-Smad2,p-Smad3,MMP-2,MMP-2 in ICS Ⅱ-L group and ICS Ⅱ-H group was significantly decreased(P<0.05)when compared with model group,and the expression of IκB-α protein was significantly increased(P<0.05).Conclusion: ICS Ⅱ inhibits the intimal hyperplasia of carotid after balloon injury of rats by the pathway of NF-κB and TGF-β1/Smads pathway.