胡黄连苷Ⅱ调节HMGB1/RAGE信号通路对冠心病大鼠内皮细胞损伤的影响

Effect of PicrosideⅡon endothelial cell damage in rats with coronary heart disease by regulating HMGB1/RAGE signaling pathway

目的:初步探讨胡黄连苷Ⅱ(P-Ⅱ)调节高迁移率族蛋白B1(HMGB1)/晚期糖基化终末产物受体(RAGE)信号通路对冠心病(CHD)大鼠内皮细胞损伤的影响。方法:SPF级SD大鼠随机分为control组、CHD组、P-Ⅱ低、中、高剂量组、P-Ⅱ高剂量+DEX组(P-Ⅱ高剂量+HMGB1/RAGE信号通路激活剂DEX),除control组外,采用高脂饮食联合腹腔注射垂体后叶素法建立CHD大鼠模型,灌胃或腹腔注射相应药物,1次/d,连续4周。生化分析仪分析大鼠血脂水平;ELISA检测血清血管内皮损伤标志物一氧化氮(NO)、内皮素(ET)-1、血管紧张素Ⅱ(AngⅡ)及血清和冠状动脉组织TNF-α、IL-1β、IL-6炎症因子水平;试剂盒检测大鼠冠状动脉组织活性氧(ROS)、谷胱甘肽过氧化物酶(GSH-Px)水平;HE染色观察大鼠冠状动脉组织病理损伤;TUNEL染色观察血管内皮细胞凋亡;Western blot检测冠状动脉组织中血管内皮生长因子(VEGF)、HMGB1、RAGE蛋白表达。结果:相较于control组,CHD组大鼠TC、TG、LDL-C、ET-1、AngⅡ、TNF-α、IL-1β、IL-6、ROS、内皮细胞凋亡率及HMGB1、RAGE表达显著升高,HDL-C、NO、GSH-Px及VEGF表达显著降低(P<0.05);相较于CHD组,P-Ⅱ低、中、高剂量组TC、TG、LDL-C、ET-1、AngⅡ、TNF-α、IL-1β、IL-6、ROS、内皮细胞凋亡率及HMGB1、RAGE表达显著降低,HDL-C、NO、GSH-Px及VEGF表达显著升高(P<0.05);HMGB1/RAGE信号通路激活剂DEX可减弱P-Ⅱ对CHD大鼠内皮细胞损伤的保护作用。结论:P-Ⅱ能有效减轻CHD大鼠内皮细胞损伤,其作用机制可能与抑制HMGB1/RAGE通路激活有关。

Objective:To investigate effect of PicrosideⅡ(P-Ⅱ)on endothelial cell damage in rats with coronary heart disease(CHD)by regulating high mobility group box 1 protein(HMGB1)/receptor for advanced glycation end products(RAGE)signaling pathway.Methods:SPF grade SD rats were randomly separated into control group,CHD group,P-Ⅱlow,medium,high doses groups and P-Ⅱhigh dose+DEX group(P-Ⅱhigh dose+HMGB1/RAGE signaling pathway activator DEX).Except for control group,CHD rat model was established by high-fat diet combined with intraperitoneal injection of pituitrin,corresponding drugs were administered by gavage or intraperitoneal injection,once a day for 4 consecutive weeks.Biochemical analyzer was applied to measure blood lipid levels;ELISA was applied to detect levels of serum nitric oxide(NO),endothelin(ET)-1,angiotensinⅡ(AngⅡ),TNF-α,IL-1βand IL-6;kits were applied to detect levels of reactive oxygen species(ROS)and glutathione peroxidase(GSH-Px)in coronary artery tissue;HE staining was applied to observe pathological damage in coronary artery tissue;TUNEL staining was applied to observe apoptosis of vascular endothelial cells;Western blot was applied to detect expressions of vascular endothelial growth factor(VEGF),HMGB1 and RAGE proteins in coronary artery tissue.Results:Compared with control group,levels of TC,TG,LDL-C,ET-1,AngⅡ,TNF-α,IL-1β,IL-6,ROS,endothelial cell apoptosis rate,and expressions of HMGB1 and RAGE in rats in CHD group were significantly increased,levels of HDL-C,NO,GSH-Px,and expression of VEGF were significantly reduced(P<0.05);compared with CHD group,levels of TC,TG,LDL-C,ET-1,AngⅡ,TNF-α,IL-1β,IL-6,ROS,endothelial cell apoptosis rate,and expressions of HMGB1 and RAGE in P-Ⅱlow,medium and high doses groups were obviously reduced,levels of HDL-C,NO,GSH-Px,and expression of VEGF were obviously increased(P<0.05);HMGB1/RAGE signaling pathway activator DEX was able to attenuate protective effect of P-Ⅱon endothelial cell damage in CHD rats.Conclusion:P-Ⅱcan effectively alleviate endothelial cell damage in CHD rats,whose mechanism may be related to inhibition of HMGB1/RAGE pathway activation.