摘要
目的观察芪丹通脉片对ApoE^-/-小鼠动脉粥样硬化易损斑块TLR4/NF-κB信号通路的影响。方法采用右侧颈总动脉外置管术(PCCP)加高脂喂养制备易损斑块模型小鼠。将手术成功的60只小鼠随机分为模型组(B组)、黄芪甲苷组[C组,黄芪甲苷40 mg/(kg·d)]、丹参酮ⅡA组[D组,丹参酮ⅡA 90 mg/(kg·d)]、益气活血组[E组,黄芪甲苷40 mg/(kg·d)+丹参酮ⅡA 90 mg/(kg·d)]、芪丹通脉片组[F组,芪丹通脉片浸膏干粉700 mg/(kg·d)],每组12只;同时设12只ApoE^-/-小鼠为对照组(A组)。于手术后当天开始,各药物干预组按上述剂量分别溶于同等体积生理盐水灌胃,A、B组给予同等体积生理盐水灌胃,每日1次。A组给予普通饲料,其余五组给予高脂饲料。共计12周。常规HE染色检测各组小鼠动脉内斑块情况,常规生化法检测TC、TG、HDL、LDL;ELISA法检测TNF-α、IL-6、C-反应蛋白(CRP)表达水平;RT-PCR检测Toll样受体4(TLR4)、核转录因子(NF-κB)、基质金属蛋白酶-9(MMP-9)mRNA的表达量及Western-Blot法检测斑块中TLR4、NF-κB蛋白相对表达水平。结果HE结果显示:A组小鼠动脉管腔内壁光滑,无斑块附着;B组小鼠可见大量斑块沉积于动脉内壁,斑块内可见到较大的脂质池,斑块结构较不稳定;与B组比较,C组及D组小鼠动脉内斑块减小,斑块内脂质池缩小,斑块结构趋向稳定,其中C、D两组比较,D组小鼠动脉斑块结构较C组稳定;E、F组小鼠动脉内壁基本光滑,少量斑块附着,结构较B、C、D组稳定。与A组比较,B组TC、TG、LDL、TNF-α、IL-6、CRP水平升高,TLR4、NF-κB、MMP-9 mRNA及TLR4、NF-κB蛋白表达水平亦升高(P<0.01),HDL水平降低(P<0.01)。与B组比较,C组TC降低,D、E、F组TC、TG、LDL水平及NF-κB、MMP-9 mRNA表达量均降低,E、F组HDL水平升高(P<0.05,P<0.01);C、D、E、F组TNF-α、IL-6、CRP水平降低,TLR4 mRNA及TLR4、NF-κB蛋白表达亦降低(P<0.05,P<0.01)。与C组比较,D组TC降低,E、F组TC、LDL水平降低,F组TG水平降低,HDL水平升高(P<0.05,P<0.01);D、E、F组TNF-α、IL-6、CRP水平降低,NF-κB、MMP-9 mRNA表达量及TLR4、NF-κB蛋白表达水平亦降低(P<0.05,P<0.01),E、F组TLR4 mRNA表达量降低(P<0.01);与D组比较,E、F组TNF-α、IL-6、CRP水平、MMP-9 mRNA表达量及TLR4、NF-κB蛋白表达水平降低(P<0.05,P<0.01),E组TG、LDL水平及NF-κB mRNA表达量降低(P<0.05),F组TLR4 mRNA表达量降低(P<0.05)。结论芪丹通脉片可能通过抑制TLR4/NF-κB信号通路,降低TNF-α、IL-6、CRP水平及MMP-9 mRNA的表达量而发挥抗易损斑块的作用。其中丹参酮ⅡA抑制易损斑块的作用可能优于黄芪甲苷,益气活血药单体配伍使用具有明显的协同作用。
Objective To observe the effect of Qidan Tongmai Tablet(QDTMT)on TLR4/NF-κB signaling pathway in ApoE-/-Mice with atherosclerotic vulnerable plaque.Methods Unstable plaque model mice were prepared by perivascular carotid collar placement(PCCP)plus high fat diet feeding.Totally 60 successfully operated mice were randomly divided into model group(group B),AstragalosideⅣgroup(group C,AstragalosideⅣ40 mg·kg^-1·d^-1),TanshinoneⅡA group(group D,TanshinoneⅡA 90 mg·kg^-1·d^-1),invigorating qi and activating blood group(group E,AstragalosideⅣ40 mg·kg^-1·d^-1+TanshinoneⅡA 90 mg·kg^-1·d^-1)and QDTMT group(group F,QDTMT extract dry powder 700 mg·kg^-1·d^-1),12 in each group.At the same time,12 ApoE^-/-mice were also set as the control group(group A).Form the day of operation,the drugs in each intervention group were dissolved in the same volume of physiological saline to give gastric gavage.The mice in group A and group B were given the same volume normal saline,once a day.The mice in group A were given normal diet,and the mice in the other five groups were given high-fat diet for 12 weeks.The Intra-arterial plaques in each group of mice was detected by HE staining.The levels of total cholesterol(TC),triglycerides(TG),high-density lipoprotein(HDL)and low-density lipoprotein(LDL)were analyzed,the expression levels of inflammatory factors TNF-α,IL-6 and C-reactive protein(CRP)were detected by ELISA,and the expression levels of Toll like receptor 4(TLR4)and NF-κB factors were detected by real-time PCR and Western Blot,the matrix metalloproteinase-9(MMP-9)mRNA level were detected by real-time PCR.Results HE results displayed:The inner wall of arterial lumen of group A was smooth and no plaque was attached;in group B,a large number of plaques were deposited on the inner wall of the artery,and a large lipid pool was observed in the plaque,and the plaque structure was unstable;compared with group B,the intra-arterial plaques in group C and group D decreased,the lipid pool in the plaque narrowed,and the plaque structure tends to be stable.Among them,the C and D groups were compared,and the arterial plaque structure of group D was stable compared with group C;In the E and F groups,the inner wall of the artery was basically smooth,a small amount of arterial plaque adhered,and the structure was more stable than group B,C and D.Compared with group A,the levels of TC,TG,LDL,TNF-α,IL-6 and CRP were increased in group B,and the expression levels of TLR4,NF-κB,MMP-9 were also increased(P<0.01),however,the level of HDL was decreased(P<0.01).Compared with group B,the level of TC was decreased in group C,the levels of TC,TG,LDL and the expression levels of NF-κB and MMP-9 were decreased in groups D,E and F.The levels of TNF-α,IL-6,CRP and the expression levels of TLR4 and NF-κB were decreased in groups C,D,E and F(P<0.05,P<0.01),but the level of HDL was increased in group E and F(P<0.05,P<0.01).Compared with group C,the level of TC was decreased in group D,the levels of TC and LDL were also decreased in group E and F.The level of TG was decreased in group F,but the level of HDL was increased(P<0.05,P<0.01).The levels of TNF-α,IL-6,CRP and the expression levels of TLR4,NF-κB,MMP-9 were also decreased compared with group C(P<0.05,P<0.01).Compared with group D,the levels of TNF-α,IL-6,CRP,MMP-9 mRNA and the protein expression levels of TLR4,NF-κB were decreased in group E and F(P<0.05,P<0.01),the levels of TG,LDL and the expression level of NF-κB mRNA were decreased in group E(P<0.05).In addition,the expression level of TLR4 mRNA was decreased in group F(P<0.05).Conclusions QDTMT may play an anti-vulnerable plaque role through inhibiting TLR4/NF-κB signaling pathway and reducing TNF-α,IL-6,CRP level and the MMP-9 mRNA expression.The results show that the role in stabilizing plaques of tanshinoneⅡA may be superior to astragalosideⅣ,and the most significant role was made by the combination of qi-invigorating and blood-activating herbs.
作者
王南丁
王文杰
王新冰
李成龙
王志梅
郝伟
马振
WANG Nan-ding;WANG Wen-jie;WANG Xin-bing;LI Cheng-long;WANG Zhi-mei;HAO Wei;MA Zhen(Department of Cardiovascology,Xi'an Traditional Chinese Medicine Hospital,Xi'an,710021;First Clinical Medical College of Shaanxi University of Chinese Medicine,Shaanxi,712046)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2019年第8期971-977,共7页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81503444)
关键词
益气活血
易损斑块
动脉粥样硬化
Toll样受体4
芪丹通脉片
invigorating qi and activating blood
vulnerable plaque
atherosclerosis
Toll-like receptors 4
Qidan Tongmai Tablet
