基于细胞焦亡NF-κB信号通路探讨黄连解毒汤干预动脉粥样硬化斑块的作用机制  

The Effect of Huanglian Jiedu Decoction on Atherosclerosis Based on NF-κB Signaling Pathway of Pyroptosis

作　　者：刘永瑞[1] 唐娜 王林[1] 叶伯鑫[1] 刘南[1] LIU Yongrui;TANG Na;WANG Lin;YE Boxin;LIU Nan(First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院,广州510405 [2]广州市白云区三源里街社区卫生服务中心

出　　处：《中西医结合心脑血管病杂志》2022年第23期4265-4273,共9页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：广东省中医药局科研基金项目。

摘　　要：目的构建新西兰兔动脉粥样硬化模型,从细胞焦亡角度研究黄连解毒汤对动脉粥样硬化的影响。方法将60只雄性新西兰兔随机分为空白组、模型组、黄连解毒汤低剂量组、黄连解毒汤中剂量组、黄连解毒汤高剂量组、VX-765组,每组10只。高脂饲料喂养联合肌肉注射脂多糖(LPS)及腹腔注射维生素D_(3)的复合方法进行造模。除空白组外,其余5组均给予复合方法进行造模。从第8周开始黄连解毒汤低剂量组、黄连解毒汤中剂量组、黄连解毒汤高剂量组分别给予黄连解毒汤每天0.75 g/kg、1.50 g/kg和3.00 g/kg灌胃,VX-765组给予氯喹[VX-765溶于二甲基亚砜(DMSO)溶液中]腹腔注射,空白组与模型组每天给予等容生理盐水灌胃,连续干预16周后,用3%戊巴比妥钠30 mg/kg腹腔注射麻醉,采用腹主动脉取血休克法处死新西兰兔,分离主动脉组织。采用苏木精-伊红(HE)染色法观察新西兰兔主动脉斑块情况,酶联免疫吸附法(ELISA)检测新西兰兔血清炎性因子水平,免疫组化(IHC)法、实时荧光定量聚合酶链式反应(RT-qPCR)法、免疫蛋白印迹(Western Blot)法评估细胞焦亡水平。结果HE染色结果显示,模型组斑块形成明显,黄连解毒汤各干预组及VX-765组主动脉粥样硬化斑块较模型组明显减少,且以黄连解毒汤高剂量组与VX-765组疗效最佳。ELISA检测炎性因子结果提示,与空白组比较,模型组C反应蛋白(CRP)、白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)、转化生长因子-β(TGF-β)、IL-18水平升高(P<0.01);与模型组比较,黄连解毒汤中剂量组、黄连解毒汤高剂量组、VX-765组CRP、IL-1β、TNF-α、TGF-β、IL-18水平下降(P<0.01);与黄连解毒汤中剂量组,黄连解毒汤高剂量组与VX-765组CRP、IL-1β、TNF-α、TGF-β、IL-18水平下降(P<0.01),且VX-765组下调IL-1β、IL-18水平较黄连解毒汤高剂量组更为明显(P<0.01)。IHC法结果显示,与空白组比较,模型组IL-1β、Objective To establish New Zealand rabbit model of atherosclerosis and study the effect of Huanglian Jiedu Decoction on atherosclerosis.Methods Sixty male New Zealand rabbits were randomly divided into blank group,model group,Huanglian Jiedu Decoction low-dose group,medium-dose group,high-dose group,and VX-765 group,with 10 rabbits in each group.High fat diet was fed with intramuscular injection of lipopolysaccharide(LPS)and intraperitoneal injection of vitamin D_(3).Except the blank group,the other 5 groups were given the compound method for modeling.From the 8th week,Huanglian Jiedu Decoction low-dose group,medium-dose group,and high-dose group were given 0.75 g/kg,1.50 g/kg,and 3.00 g/kg of Huanglian Jiedu Decoction by gavage,respectively.The VX-765 group was given chloroquine intraperitoneal injection,blank group and model group were given equal volume normal saline gavage every day.After 16 weeks of continuous intervention,the rabbits were anesthetized,and the aorta tissues were isolated.Hematoxylin and eosin(HE)staining was used to observe the aortic plaque of New Zealand rabbits.The levels of inflammatory factors in serum were detected by enzyme linked immunosorbent assay(ELISA).The levels of pyroptosis were evaluated by immunohistochemistry(IHC),real-time fluorescence quantitative PCR(RT-qPCR)and Western Blot.Results The results of HE staining showed that the model group had obvious plaque formation,and the aortic atherosclerotic plaque in the Huanglian Jiedu Decoction and VX-765 group was significantly less than that in the model group,and the Huanglian Jiedu Decoction high-dose group and VX-765 group were the best ones.The results of ELISA showed that compared with the blank group,the levels of C-reactive protein(CRP),interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),transforming growth factor-β(TGF-β),and IL-18 in the model group were high(P<0.01).Compared with model group,the levels of CRP,IL-1β,TNF-α,TGF-β,and IL-18 in medium-dose group,high-dose group and VX-765 group were less(P<0.01).Compa

关 键 词：动脉粥样硬化 黄连解毒汤 核转录因子-κB号通路 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]