| Objective:The purpose of this study was to explore the effect and molecular mechanism of Buyang Huanwu decoction(BHD)on atherosclerosis(AS).In this study,the AS model of apolipoprotein E gene knockout(ApoE-/-)mice was established by high-fat diet,and interfered with the prescription of replenishing qi and activating blood circulation,so as to provide theoretical and experimental basis for the treatment of AS with BHD.Methods:40 male ApoE-/-mice with C57BL/6J background(8 weeks old)were randomly di-vided into four groups:blank group(n=10),model group(n=10),BHD group(n=10)and Atorvastatin(Ato)group(n=10).The mice in the blank group were fed with normal diet,the model group,BHD group and Ato group were given high fat diet,the blank group and model group were given the same volume of normal saline,the BHD group was given BHD(18.53g/kg)and the Ato group was given Ato(1.3mg/kg)for 8 weeks.The changes of body weight of mice were recorded every week during the experiment.At the end of the experiment,the levels of serum total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)were measured by automatic biochemical analyzer,and the plaque size of aortic arch was observed by oil red O staining and the proportion of lipid droplets was analyzed.The serum concentrations of interleukin-1?(IL-1?),IL-6,tumor necrosis factor-a(TNF-?),IL-4,IL-10 and transforming growth factor-?(TGF-?)were measured by ELISA.The protein expression levels of inducible nitric oxide synthase(iNOS),arginase-1(Arg-1)and macrophage polarization hub pyruvate kinase M2(PKM2)/hypoxia inducible factor-1?(HIF-1?)in M1 and M2 macrophages were detected by Western-blot,and the gene expression levels of aortic cytokines IFN-y,IL-1?,IL-6,TNF-?,IL-4,IL-10 and TGF-? were detected by RT-PCR.Results:1.The results of blood lipid detection:compared with the blank group,the four items of blood lipid TC,TG,LDL-C and HDL-C in the model group were significantly increased(P<0.001).Compared with the model group,TC,TG,LDL-C and HDL-C in BHD group and Ato group were significantly decreased(P<0.001).2.The pathological results showed that the percentage of lipid droplets in the model group was significantly higher than that in the blank group(P<0.001),while that in the BHD group and the Ato group was significantly lower than that in the model group(P<0.05).3.ELISA results:compared with the blank group,the expression of IL-1?,IL-6 and TNF-? in the model group increased significantly(P<0.01,P<0.05,P<0.001),while the expression of IL-4 and IL-10 decreased significantly(P<0.05).Compared with the model group,the expression of IL-1? and TNF-? in BHD group was significantly decreased and the expression of IL-4 and IL-10 was significantly increased(P<0.01).The expression of IL-4 in Ato group was significantly higher than that in model group(P<0.01).But the expression of TGF-? had no significant change.4.Western-blot results:compared with the blank group,the protein expression levels of iNOS,PKM2 and HIF-1 a in the model group were significantly increased(P<0.01),while the expression of Arg-1 was significantly decreased(P<0.01).Compared with the model group,the protein expression levels of iNOS,PKM2 and HIF-1? in Ato group and BHD group were significantly decreased(P<0.01 P<0.01,P<0.05 or P<0.001,P<0.01,P<0.01),and the expression of Arg-1 was significantly increased(P<0,01).5.RT-PCR results:compared with the blank group,the gene expression levels of IFN-?,IL-1?,IL-6,TNF-?,GLUT1,PDK1 and LDHA in the model group were significantly increased(P<0.05),while IL-4,IL-10 and TGF-? were significantly decreased(P<0.05).Compared with the model group,the gene expression levels of IFN-?,IL-1?,IL-6,TNF-?,GLUT1,PDK1 and LDHA in BHD group and Ato group were significantly decreased(P<0.05,P<0.01,P<0.001),while IL-4 and IL-10 were significantly increased(P<0.05,P<0.01),but TGF-? had no significant change(P>0.05).Conclusion:BHD can reduce the body weight and serum lipids(TC,TG,LDL-C)of AS model mice,reduce the proportion of aortic plaque lipid droplets' and also reduce the level of inflammatory factors and increase the level of anti-inflammatory factors.Its mechanism of action may be related to regulating the expression of M1/M2 macrophage polarization hub PKM2/HIF-1? and its downstream glycolytic enzymes,thereby regulating the polarization balance of M1/M2 macrophages,biasing macrophage differentiation towards the M2 phenotype,achieving anti-inflammatory,promote plaque stability and delay the progression of AS. |
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