Exploring The Mechanism Of Yiqiliangxueshengji Prescription In Intervening Endothelial Cell Injury And Repair Based On NF-κB Pathwa

Background:Percutaneous coronary intervention(PCI)is an indispensable treatment for coronary heart disease,which reduces the mortality of acute myocardial infarction in coronary heart disease.From the earliest percutaneous transluminal coronary angioplasty(PTCA)to the bare metal stent(BMS)era,to the emergence of drug-eluting stents(DES)and degradable stents,PCI has been around for more than 40 years,which technology continues to break through.But there are still problems such as in-stent restenosis and long-term thrombosis.After PCI,whether balloon dilatation or stent implantation,it will bring inevitable mechanical damage to the blood vessel wall,leading to endothelial cell damage.The repair of endothelium after injury is still a difficult problem in PCI.Endothelial injury initiates an inflammatory response,which is accompanied by the entire process of endothelial repair.Previous studies have shown that Yiqi Liangxue-shengji decoction(YL)can protect the vascular endothelium.The main components of Yiqi Liangxue-shengji decoction,Huangqi and Danshen,whicn can inhibit the over-activation of NF-κB.NF-κB is involved in the regulation of various inflammatory factors and cytokines and participates in the inflammatory process.Objective:To establish a model of inflammatory injury induced by TNF-α in human umbilical artery endothelial cells(HUAECs)and explore the mechanism of YL to protect the endothelium based on NF-κB pathway.Methods:Experiment 1:The model of inflammatory injury of HUAECs induced by TNF-α was established.The study group consisted of:control group,model group,high concentration of Yiqi-Liangxue Group(HYL),low concentration of Yiqi-Liangxue Group(LYL),Atorvastatin group.Cell viability and proliferation were detected by CCK-8 assay,endothelial cell proliferation and migration were observed by cell scratch assay,and TLR4,MyDD88,NF-κB,ICAM-1,Bcl-2 mRNA and protein expression were detected by RT-PCR and Western blot.Experiment 2:The model of inflammatory injury of HUAECs induced by TNF-α was established.The study group consisted of:control group,model group,inhibitor(TAK-242)group,inhibitor(TAK-242)+high concentration of Yiqi-Liangxue group(Inhibitor+HYL).Cell viability and proliferation were detected by CCK-8 assay,endothelial cell proliferation and migration were observed by cell scratch assay,and TLR4,MyDD88,NF-κB,ICAM-1,Bcl-2 mRNA and protein expression were detected by RT-PCR and Western blot.Result:Experiment 1:CCK-8:When the concentration of the serum was 10%and 15%,compared with the model group,YL group,Atorvastatin group all enhanced the viability and proliferation of injured cells(P<0.05).HYL group had better effects than LYL group and Atorvastatin group(P<0.05),inhibitor+HYL group had better effects than inhibitor group(P<0.05).Scratch test:compared with the model group,LYL group,HYL group,Atorvastatin group.Western blot:Compared with the model group,the expressions of TLR4,MyD88,NF-κB/p65 and ICAM-1 were decreased in the LYL group,HYL group,Atorvastatin group(P<0.05),the expression of Bcl-2 was increased(P<0.5).Compared with the LYL group,the expression of NF-κB/p65 was increased in Atorvastatin group(P<0.05)when MyD88,ICAM-1 and Bcl-2 were decreased(P<0.05),MyD88 was decreased in HYL group(P<0.05)when Bcl-2 was Increased(P<0.05).Real time PCR:Compared with the model group,the expressions of TLR4,MyD88,NF-KB/p65 and ICAM-1 were decreased in LYL group,HYL group,Atorvastatin group(P<0.05),the expression of Bcl-2 was increased in the HYL group(P<0.5).Compared with the LYL group,NF-κB/p65 was increased in Atorvastatin group(P<0.05)when MyD88 and ICAM-1 was decreased(P<0.05),MyD88 and NF-KB/p65 were decreased in HYL group(P<0.05)when Bcl-2 was increased(P<0.05).Experiment 2:CCK-8:compared with the model group,inhibitor group,inhibitor+HYL group all enhanced the viability and proliferation of injured cells(P<0.05).Scratch test:compared with the model group,Inhibitor group,Inhibitor+HYL group had accelerated cell migration(P<0.05).Western blot:Compared with the model group,the expressions of TLR4,MyD88,NF-κB/p65 and ICAM-1 were decreased in the inhibitor group,Inhibitor+HYL group(P<0.05),the expression of Bcl-2 was increased(P<0.5).Compared with inhibitor group,MyD88 and ICAM-1 were decreased(P<0.05)and Bcl-2 was increased(P<0.05)in inhibitor+HYL group.Real time PCR:Compared with the model group,the expressions of TLR4,MyD88,NF-κB/p65 and ICAM-1 were decreased in Inhibitor group,Inhibitor+HYL group(P<0.05).Compared with the inhibitor group,MyD88 and ICAM-1 were decreased in the inhibitor+HYL group(P<0.05).Conclusion:Yiqi Liangxue-shengji decoction can promote the proliferation and migration of vascular endothelial cells induced by TNF-α.Yiqi Liangxue-shengji decoction inhibits the inflammatory response of vascular endothelial cells induced by TNF-α,and its mechanism may be related to the inhibition of NF-κB pathway.