| Objectives:The purpose of this study was to observe the clinical effect of Guanxinping(GXP)on elderly patients with stable coronary heart disease complicated with carotid plaques,to investigate the ameliorating effect of Guanxinping on atherosclerotic inflammatory response in ApoE-/-mice and its inhibitory effect on inflammation response of human umbilical vein endothelial cells(HUVEC).We explored its mechanism of ameliorating the inflammatory response from MAPK/NF-κB signaling pathway.Methods:In the clinical trial,100 elderly patients with stable coronary artery disease complicated with carotid plaques who met the diagnostic criteria were selected and randomly divided into 2 groups by single-blind and parallel control method.On the basis of conventional western medicine intervention,the treatment group was combined with Guanxinping tablets,and the control group was combined with Yixinshu tablets.The course of treatment was 3 months.The angina attack,ECG changes,the TCM syndrome score,carotid intima-media thickness(C-IMT),and Crouse score were recorded and analyzed before and after treatment.In vivo experiments,ApoE-/-mouse was fed with High-fat Diet to creat AS Model,and then these mice were randomly divided into model group(MOD),atorvastatin group(ATO),low-dose Guanxinping group(LGXP),high-dose Guanxinping group(HGXP).After 12 weeks of intervention,the body weight and blood lipids of the mice were detected.The structure and morphology of the aorta of ApoE-/-mice were observed by hematoxylin-eosin(HE)staining.The lipid deposition in the aorta of the ApoE-/-mice was observed by oil red staining.The mRNA expressions of inflammatory factors TNF-α,IL-6,IL-1β,VCAM-1 and ICAM-1 in mouse aorta were detected by qPCR.The expressions of p-MAPK,p-NF-κB and JNK in the aorta of mice were detected by immunohistochemistry;the inflammatory levels of TNF-α,IL-1β and IL-6 in the serum of mice were detected by ELISA;the protein levels of inflammatory factors such as TNF-α,,IL-6,IL-1β,VCAM-1 and ICAM-1 were detected by Western blot,and the expressions of MAPK,NF-κB,JNK,ERK and other proteins were further detected.In vitro experiments,lipopolysaccharide(LPS)was used to induce HUVEC to establish a cell inflammation injury model,and the cultured cells were divided into 5 groups:normal control group(CON),model group(MOD),atorvastatin group(ATO),low-dose Guanxinping group(LGXP),medium-dose Guanxinping group(MGXP),and high-dose Guanxinping group(HGXP).Cells and supernatants were collected 24 hours after drug addition.The inflammatory levels of TNF-α,IL-1β and IL-6 in the supernatant of cells were detected by ELISA.The total RNA of cells in each group was extracted.The mRNA expressions of inflammatory factors TNF-α,IL-6,IL-1β,VCAM-1 and ICAM-1 in cells of each group were detected by qPCR.The expressions of inflammatory factors and key proteins in MAPK/NF-κB signaling pathway were detected by Western blot.Results:In the clinical trial,Guanxinping could improve the symptom scores of angina pectoris in elderly patients with stable coronary artery disease complicated with carotid plaques(P<0.05),the effect of electrocardiogram change was better than that of the control group(P<0.05),and the scores of TCM syndromes were better than those of the control group(P<0.05).Guanxinping could decrease the C-IMT score and Crouse score(P<0.05).In vivo experiments,compared with the MOD group,the levels of TG,TC,and LDL-c in the GXP group were significantly lower(P<0.05),while the HDL-c level was significantly higher than that in the model group(P<0.05),suggesting that GXP could significantly ameliorate lipid metabolism in ApoE-/-mice induced by high-fat diet;both ATO and GXP significantly reduced lipid deposition in the aorta compared with the MOD group.Pathological sections showed that the aortic endothelium of mice in the MOD group was significantly thickened and infiltrated by inflammatory cells,which was significantly ameliorated after GXP and ATO intervention.The results of serum ELISA showed that compared with the MOD group,the inflammatory levels of TNF-α,IL-1β and IL-6 in the GXP intervention group were significantly decreased(P<0.05),which was consistent with the mRNA expression of inflammatory factors in the qPCR results.Immunohistochemical results showed that the protein expressions of p-MAPK,p-NF-κB and JNK in the MOD group were increased,whereas the protein expression levels were decreased after GXP intervention.The results of Western blot indicated that the expressions of MAPK,NF-κB,JNK and ERK in the MOD group were increased,which were significantly inhibited after GXP intervention(P<0.05).In vitro experiments,the LPS-induced HUVEC cell inflammation model was used,the levels of cellular inflammatory factors TNF-α,IL-1β,IL-6,ICAM-1,and VCAM-1 were significantly increased,which was consistent with the results of the mRNA and protein levels(P<0.05).After 24 hours of treatment with ATO and GXP,the levels of cellular inflammatory factors TNF-α,IL-6,IL-1β,VCAM-1,and ICAM-1 were decreased(P<0.05),and the mRNA and protein levels showed consistent results(P<0.05).Not only that,the expressions of MAPK,NF-κB,JNK and ERK were significantly increased in the MOD group,and were significantly decreased after GXP and ATO intervention(P<0.05).Conclusions:1.Guanxinping can improve the main TCM symptom score,frequency and duration of angina pectoris in elderly patients with stable coronary artery disease complicated with carotid plaque.Guanxinping can ameliorate myocardial ischemia.Guanxinping can reduce carotid intima-media thickness and plaque thickness.2.Guanxinping can reduce serum lipid levels in ApoE-/-mice,reduce lipid deposition in the aorta,and inhibit the expression of inflammatory cytokines and key proteins in the MAPK/NF-κB signaling pathway.Guanxinping inhibits the inflammatory response of atherosclerosis in ApoE-/-mice by inhibiting the activation of MAPK/NF-κB pathway.3.Guanxinping can inhibit the levels of inflammatory factors and the expression of key proteins in the MAPK/NF-κB signaling pathway in the LPS-induced HUVEC inflammation model.Guanxinping inhibits HUVEC inflammatory response by inhibiting the activation of MAPK/NF-κB signaling pathway. |
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