心痛泰颗粒对动脉粥样硬化ApoE^(−/−)小鼠ox-LDL、ICAM-1和VCAM-1表达的影响

Effects of Xin-Tong-Tai granule on expression of ox-LDL,ICAM-1 and VCAM-1 in ApoE^(−/−)mice with atherosclerosis

目的:探讨心痛泰颗粒对动脉粥样硬化ApoE^(−/−)小鼠氧化型低密度脂蛋白(ox-LDL)、细胞间黏附分子1(ICAM-1)和血管细胞黏附分子1(VCAM-1)表达的影响及其机制。方法:6~8周龄SPF级健康雄性ApoE^(−/−)小鼠72只,采用高脂饮食喂养12周进行造模,另设SPF级健康雄性C57BL/6J野生小鼠12只为对照组,予以普通饲料喂养。各组相应药物给药8周后,观察各组小鼠体质量及一般情况,采用生化试剂盒检测小鼠血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)含量,HE染色和油红O染色观察主动脉病理结构,ELISA法检测血清ox-LDL及主动脉组织中ICAM-1和VCAM-1水平,Western blot法检测主动脉NADPH氧化酶4(NOX4)、NOX亚单位p22phox、核因子抑制蛋白激酶α(IKK-α)、IKK-β和核因子κB(NF-κB)蛋白表达情况。结果:与对照组比较,模型组小鼠体质量增加(P<0.05),且毛发晦暗无光泽、局部脱落,抓起反应迟钝;血清TC、TG和LDL-C上升,HDL-C下降(P<0.05),血清ox-LDL水平上升(P<0.05),主动脉ICAM-1和VCAM-1水平升高(P<0.05),主动脉NOX4、p22phox、IKK-α、IKK-β和NF-κB蛋白表达增加(P<0.05)。与模型组比较,各用药组小鼠体质量下降(P<0.05),且毛发脱落及反应灵活程度亦有所改善;血清TC、TG和LDL-C降低,HDL-C升高(P<0.05),血清ox-LDL水平下降(P<0.05),主动脉ICAM-1和VCAM-1水平降低(P<0.05),主动脉NOX4、p22phox、IKK-α、IKK-β和NF-κB蛋白表达减少(P<0.05)。HE及油红O染色显示,模型组小鼠血管内可见典型动脉粥样硬化斑块,且红染区域分布广泛;各用药组与模型组比较,以上情况均有不同程度减轻。结论:心痛泰颗粒可减少高脂饮食诱导的ApoE^(−/−)小鼠动脉粥样硬化斑块面积,下调血清TC、TG和LDL-C水平,升高HDL-C水平,减少血清ox-LDL水平,下调主动脉ICAM-1和VCAM-1水平,抑制主动脉NOX4、p22phox、IKK-α、IKK-β和NF-κB蛋白的表达,改善动脉粥样硬化。

AIM:To investigate the effects and mechanism of Xin-Tong-Tai granule on oxidized low-density lipoprotein(ox-LDL),intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)in ApoE^(−/−)mice with atherosclerosis(AS).METHODS:A total of 72 SPF-grade healthy male ApoE^(−/−)mice aged 6~8 weeks were fed with high-fat diet for 12 weeks to replicate AS models,and 12 SPF-grade healthy male C57BL/6J wild mice were fed with ordinary diet as the control group.After the corresponding drugs were administered for 8 weeks,the body weight and general condition of mice in each group were observed.The serum levels of total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)were detected by biochemical kits.The pathological structures of aorta were observed by HE and oil red O staining.The levels of serum ox-LDL and aortic ICAM-1 and VCAM-1 were detected by ELISA.The protein levels of NADPH oxidase 4(NOX4),NOX subunit p22phox,inhibitor ofκB kinase-α(IKK-α),IKK-βand nuclear factor-κB(NF-κB)in aorta were detected by Western blot.RESULTS:Compared with control group,the mice in model group showed increased body weight(P<0.05),dull and local shedding hair,slow grasping response,increased serum TC,TG and LDL-C levels,decreased serum HDL-C level(P<0.05),increased the levels of serum ox-LDL and aortic ICAM-1 and VCAM-1(P<0.05),and increased protein expressions of NOX4,p22phox,IKK-α,IKK-βand NF-κB in aorta(P<0.05).Compared with model group,the body weight of mice in each treatment group decreased(P<0.05),the hair loss and the response flexibility were also improved.The serum levels of TC,TG and LDL-C decreased and HDL-C increased(P<0.05).The levels of serum ox-LDL and aortic ICAM-1(except the low-dose Xin-Tong-Tai granule group)and VCAM-1 decreased(P<0.05).The protein levels of NOX4,p22phox,IKK-α,IKK-βand NF-κB in aorta decreased(P<0.05).HE and oil red O staining showed that typical AS plaques could be seen in blood vessels of the model group,and the red-stained areas were widely distributed.The above lesions were alleviated to different degrees in each treatment group compared with model group.CONCLUSION:Xin-Tong-Tai granule reduces the atherosclerotic plaque area of ApoE^(−/−)mice induced by high-fat diet,decreased serum TC,TG and LDL-C levels,increased HDL-C level,decreased the levels of serum ox-LDL and aorta ICAM-1 and VCAM-1,and inhibited protein expression of NOX4,p22phox,IKK-αand IKK-βin the aorta,thereby attenuating AS.