摘要
目的基于NF-κB/NLRP3信号通路探讨通心化浊方改善冠状动脉粥样硬化性心脏病的机制。方法选取6~8周龄SPF级雄性SD大鼠48只,按随机数字表法将其分为空白组、模型组、通心化浊方低剂量组、通心化浊方中剂量组、通心化浊方高剂量组、阳性药组、BAY-11-7082组、通心化浊方高剂量+1/2BAY-11-7082组,每组6只。除空白组外,其余大鼠进行高脂饮食加脂肪乳剂灌胃和腹腔注射垂体后叶素,制造冠状动脉粥样硬化性心脏病大鼠模型。造模成功后,通心化浊方低、中、高剂量组分别以6.21、12.42、24.84 g/(kg·d)通心化浊方灌胃;阳性药组以辛伐他汀灌胃(1.8 mg/kg);BAY-11-7082组以BAY-11-7082(5 mg/kg,隔日1次)腹腔注射;通心化浊方高剂量+1/2BAY-11-7082组以每日通心化浊方[24.84 g/(kg·d)]灌胃及BAY-11-7082(2.5 mg/kg,隔日1次)腹腔注射,正常组和模型组,以10 ml/kg蒸馏水灌胃。连续给药4周后取材,检测大鼠血清中低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)、甘油三酯(TG)、白细胞介素(IL)-6、IL-1β、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)、脂联素水平;HE染色观察心肌组织的病理学改变;qRT-PCR法检测血管细胞黏附分子-1(VCAM-1)、细胞间黏附分子-1(ICAM-1)、IL-1β、单核细胞趋化蛋白-1(MCP-1)的mRNA表达;Western blot检测大鼠心肌组织NF-κB/NLRP3通路蛋白表达情况。结果与空白组比较,模型组大鼠心肌组织出现病理损伤,血清TC、TG、LDL-C、IL-6、IL-1β、MCP-1、TNF-α含量升高,脂联素水平降低,VCAM-1、ICAM-1、IL-1β、MCP-1的mRNA表达量增加,心肌组织NF-κB p65、NLRP3通路蛋白表达水平升高(P<0.01);与模型组比较,经药物干预后,大鼠心脏组织病理变化得到改善,各给药组TC、TG、LDL-C、IL-6、IL-1β、MCP-1、TNF-α水平降低(P<0.01)。通心化浊方高剂量组TG、LDL-C水平低于通心化浊方中、低剂量组,且通心化浊方中剂量组低于通心化浊方低剂量组(P<0.01);通心化浊方中、高剂量组TC水平低于通心化浊方低剂量组(P<0.01)。通心化浊方高剂量组血清IL-6、IL-1β、MCP-1、TNF-α水平低于通心化浊方低剂量组(P<0.05);通心化浊方高剂量组及通心化浊方高剂量+1/2BAY-11-7082组血清IL-6、MCP-1、TNF-α水平低于BAY-11-7082组(P<0.05)。与模型组比较,各给药组血清脂联素水平增高(P<0.01)。通心化浊方中、高剂量组血清脂联素水平高于通心化浊方低剂量组,且通心化浊方高剂量组高于通心化浊方中剂量组(P<0.01);BAY-11-7082组血清脂联素水平低于通心化浊方高剂量+1/2BAY-11-7082组(P<0.05)。与模型组比较,各给药组VCAM-1、ICAM-1、IL-1β、MCP-1的mRNA表达量下降(P<0.01)。通心化浊方中、高剂量组VCAM-1、ICAM-1、IL-1β、MCP-1的mRNA表达量低于通心化浊方低剂量组(P<0.01),且通心化浊方高剂量组低于通心化浊方中剂量组(P<0.05)。与模型组比较,各给药组心肌组织NF-κB p65、NLRP3蛋白表达降低(P<0.05)。通心化浊方高剂量组心肌组织NF-κB p65、NLRP3蛋白表达低于通心化浊方低剂量组(P<0.05)。结论通心化浊方有改善冠状动脉粥样硬化性心脏病的作用,其机制可能是降低血脂,上调脂联素表达,抑制NF-κB/NLRP3信号通路,减少IL-6、IL-1β、MCP-1、TNF-α炎症因子和VCAM-1、ICAM-1血管黏附分子的表达而减轻炎症反应,减少动脉粥样硬化的发生或延缓动脉粥样硬化的发展,从而改善冠状动脉粥样硬化性心脏病。
Objective To investigate the mechanism of Tongxin Huazhuo Prescription on coronary atherosclerotic heart disease based on NF-κB/NLRP3 signaling pathway.Methods A total of 48 SPF male SD rats aged six to eight weeks were selected and divided into blank group,model group,Tongxin Huazhuo Prescription low dose group,Tongxin Huazhuo Prescription medium dose group,Tongxin Huazhuo Prescription high dose group,positive drug group,BAY-11-7082 group,and Tongxin Huazhuo Prescription high dose+1/2 BAY-11-7082 group according to random number table method,with six rats per group.Except the blank group,the other rats were given high-fat diet plus fat emulsion and intraperitoneal injection of pituitrin to create a rat model of coronary atherosclerotic heart disease.After successful molding,Tongxin Huazhuo Prescription low,medium,and high dose groups were administrated with 6.21,12.42,and 24.84 g/(kg·d)Tongxin Huazhuo Prescription.The positive group was given Simvastatin intragastine(1.8 mg/kg).BAY-11-7082 group was given intrabitoneal injection of BAY-11-7082(5 mg/kg,once every other day).Tongxin Huazhuo Prescription high dose+1/2BAY-11-7082 group was given daily Tongxin Huazhuo Prescription(24.84 g/[kg·d])and BAY-11-7082(2.5 mg/kg,once every other day)by intrabitoneal injection;normal group and model group were given 10 ml/kg by distilled water.The serum levels of low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),triglyceride(TG),interleukin(IL)-6,IL-1β,monocyte chemoattractant protein-1(MCP-1),tumor necrosis factor-α(TNF-α),and adiponectin were measured after four weeks of continuous administration.The pathological changes of myocardial tissue were observed by HE staining.The mRNA expressions of vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),IL-1β,and monocyte chemotactic protein-1(MCP-1)were detected by qRT-PCR.Western blot analysis of NF-αB/NLRP3 pathway protein expression in rat myocardium.Results Compared with the blank group,the myocardial tissue of rats in the model group showed pathological injury,the contents of TC,TG,LDL-C,IL-6,IL-1β,MCP-1,and TNF-αin serum were increased,the level of adiponectin was decreased,and the mRNA expression levels of VCAM-1,ICAM-1,IL-1β,and MCP-1 were increased.The expression levels of NF-κB p65 and NLRP3 pathway protein in myocardial tissue were increased(P<0.01).Compared with the model group,after drug intervention,the pathological changes of rat heart tissue were improved,and the levels of TC,TG,LDL-C,IL-6,IL-1β,MCP-1,and TNF-αwere decreased in each drug administration group(P<0.01).The levels of TG and LDL-C in Tongxin Huazhuo Prescription high dose group were lower than those in Tongxin Huazhuo Prescription middle and low dose groups,and those in Tongxin Huazhuo Prescription middle dose group were lower than those in Tongxin Huazhuo Prescription low dose group(P<0.01).The TC level in Tongxin Huazhuo Prescription middle and high dose groups was lower than that in Tongxin Huazhuo Prescription low dose group(P<0.01).The levels of IL-6,IL-1β,MCP-1,and TNF-αin Tongxin Huazhuo Prescription high dose group were lower than those in Tongxin Huazhuo Prescription low dose group(P<0.05).The serum levels of IL-6,MCP-1,and TNF-αin Tongxin Huazhuo Prescription high dose group and Tongxin Huazhuo Prescription high dose+1/2BAY-11-7082 group were lower than those in BAY-11-7082 group(P<0.05).Compared with the model group,the serum adiponectin level in each administration group was increased(P<0.01).The serum adiponectin level of Tongxin Huazhuo Prescription medium and high dose groups was higher than that of Tongxin Huazhuo Prescription low dose group,and that of Tongxin Huazhuo Prescription high dose group was higher than that of Tongxin Huazhuo Prescription medium dose group(P<0.01).The serum adiponectin level of BAY-11-7082 group was lower than that of Tongxin Huazhuo Prescription high dose+1/2BAY-11-7082 group(P<0.05).Compared with model group,the mRNA expression levels of VCAM-1,ICAM-1,IL-1β,and MCP-1 in each administration group were decreased(P<0.01).The mRNA expression levels of VCAM-1,ICAM-1,IL-1β,and MCP-1 in Tongxin Huazhuo Prescription middle and high dose groups were lower than those in Tongxin Huazhuo Prescription low dose group(P<0.01),and those in Tongxin Huazhuo Prescription high dose group were lower than those in Tongxin Huazhuo Prescription medium dose group(P<0.05).Compared with model group,the expressions of NF-κB p65 and NLRP3 protein in myocardial tissue of all drug administration groups were decreased(P<0.05).The protein expressions of NF-κB p65 and NLRP3 in myocardial tissue of Tongxin Huazhuo Prescription high dose group were lower than those of Tongxin Huazhuo Prescription low dose group(P<0.05).Conclusion Tongxin Huazhuo Prescription can ameliorate coronary atherosclerotic heart disease,and its mechanism may be to reduce blood lipid,up-regulate adiponectin expression,inhibit NF-κB/NLRP3 signaling pathway,reduce IL-6,IL-1β,MCP-1,TNF-αinflammatory factors,VCAM-1,ICAM-1 vascular adhesion molecules and reduce inflammation.Reduce the occurrence of atherosclerosis or delay the development of atherosclerosis,thereby improving coronary atherosclerotic heart disease.
作者
谢珍聪
孙广瀚
孙咪
徐洋
郭栋
XIE Zhencong;SUN Guanghan;SUN Mi;XU Yang;GUO Dong(The First School of Clinical Medicine,Shandong University of Traditional Chinese Medicine,Shandong Province,Jinan 250000,China;Teacher Development Center,Shandong University of Traditional Chinese Medicine,Shandong Province,Jinan 250000,China;Pharmaceutical Research Institute,Shandong University of Traditional Chinese Medicine,Shandong Province,Jinan 250000,China)
出处
《中国医药导报》
2025年第6期42-49,共8页
China Medical Herald
