基于PI3K/AKT/eNOS通路研究芪丹通脉片对ApoE-/-小鼠动脉粥样硬化易损斑块的影响OACSTPCD

Objective:To explore the effect of Qidantong Maipian on vulnerable atherosclerotic plaques in ApoE-/-mice based on the pathway of phosphatidylinositol-3-hydroxykinase(PI3K)/protein kinase B(AKT)/endothelial nitric oxide synthase(eNOS).Meth-od:Twelve male ApoE-/-mice of SPF grade were randomly selected as the control group(Group A),while the remaining mice were subjected to common carotid artery catheterization combined with high-fat diet to establish a mouse vulnerable plaque model.Sixty successfully modeled mice were randomly divided into a model group(Group B),an astragalosideⅣgroup(Group C,40 mg·kg-1),a tanshinone ⅡA group(Group D,90 mg·kg-1),an astragalosideⅣ+tanshinoneⅡA group(Group E),and a Qidan Tongmai tab-let group(Group F,700 mg·kg-1),with 12 mice in each group.The corresponding drugs were administered orally for 12 weeks,once a day.The control group and model group were given equal volumes of physiological saline.After the last gavage,the mice were eutha-nized and the right common carotid artery was separated.TUNEL staining was used to observe cell apoptosis,and immunofluorescence staining was used to detect the expression level of CD68 and α-actin,RT-PCR detection of eNOS mRNA expression,Western Blot detection of p-PI3K,p-AKT,eNOS,and matrix metalloproteinase-9(MMP-9)protein expression levels.Results:TUNEL results showed that group A had almost no apoptosis in smooth muscle cells,while group B had the highest apoptosis in smooth muscle cells;Compared with group B,the apoptosis of smooth muscle cells in the C-F group was significantly reduced;E.The apoptosis of smooth muscle cells in group F was reduced compared to groups C and D.CD68 and α-actin immunofluorescence results showed that Group A had almost no macrophages,while the number of smooth muscle cells was the highest;Compared with group A,the number of macropha-ges in group B significantly increased,while the number of smooth muscle cells significantly decreased;Compared with group B,the C-F group showed a significant decrease in macrophages and a significant increase in smooth muscle cells;Compared with C and D groups,E and F groups showed a significant decrease in macrophages and a significant increase in smooth muscle cells.The RT-PCR results showed that compared with group A,the expression of eNOS mRNA in group B was significantly reduced(P＜0.05);Compared with Group B,the expression of eNOS mRNA in Group C-F was significantly increased(P＜0.05);Compared with groups C and D,the expression of eNOS mRNA in groups E and F was significantly increased(P＜0.05).Western blot results showed that compared with group A,the expression of p-PI3K,p-AKT,and eNOS proteins in group B was significantly reduced,while the expression of MMP-9 protein was significantly increased(P＜0.05);Compared with Group B,the expression of p-PI3K,p-AKT,and eNOS pro-teins in Group C-F was significantly increased,while the expression of MMP-9 protein was significantly reduced(P＜0.05);Com-pared with groups C and D,the expression of p-PI3K,p-AKT,and eNOS proteins in groups E and F was significantly increased,while the expression of MMP-9 protein was significantly reduced(P＜0.05).Conclusion:Qidan Tongmai Tablet can stabilize vulnera-ble atherosclerotic plaque,and its mechanism may be related to the activation of PI3K/AKT/eNOS signaling pathway.