四妙勇安汤对ApoE~(-/-)动脉粥样硬化小鼠IL-6和MCP-1的影响
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摘要
目的:研究四妙勇安汤对高脂饲料喂养的ApoE基因敲除小鼠动脉粥样硬化炎症反应的干预作用。方法:采用高脂饲料喂养ApoE~(-/-)小鼠14周制备动脉粥样硬化模型,成模后随机分为5组:模型组,阳性药组(阿托伐他汀,2.57 mg·kg~(-1)),四妙勇安汤高剂量组(34.9 g·kg~(-1))、四妙勇安汤中剂量组(23.14 g·kg~(-1))和四妙勇安汤低剂量组(11.61 g·kg~(-1)),分别灌胃给药12周,同时以C57BL/6小鼠为正常对照组,给予去离子水灌胃12周。采用生化仪检测三酰甘油(TG)、总胆固醇(TC)、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染色观察主动脉血管形态,并测量主动脉内膜厚度(IT)、中膜厚度(MT)、血管管腔面积(LA)、粥样斑块面积(PA)、内中膜厚度比(IT/MT)、管腔狭窄率(PA/LA);采用流式高通量多因子检测技术检测血清中白介素-6(IL-6)、单核细胞趋化蛋白-1(MCP-1)炎症细胞因子表达。结果:与对照组相比,模型组小鼠血清TC、TG、LDL-C明显提高,HDL-C明显降低,PA、IT及IT/MT明显升高(P<0.01)。与模型组比较,四妙勇安汤能够明显降低ApoE~(-/-)小鼠主动脉PA、IT及IT/MT(P<0.01),并明显降低IL-6及MCP-1表达(P<0.01、P<0.05),但对血脂水平无明显的影响。结论:四妙勇安汤可以通过抑制炎症反应防治动脉粥样硬化,而不依赖血脂水平的变化。
Objective:To study the intervention effect of Simiao Yongan decoction on atherosclerotic inflammatory response in ApoE knockout mice fed with high fat diet.Methods:ApoE~(-/-)mice were fed with high-fat diet for 14 weeks to prepare atherosclerosis model.After modeling,they were randomly divided into 5 groups:model group,positive drug group(atorvastatin,2.57 mg·kg~(-1)),Simiao Yongan decoction high dose group(34.9 g·kg~(-1)),Simiao Yongan decoction medium dose group(23.14 g·kg~(-1))and Simiao Yongan low dose group(11.61 g·kg~(-1)),respectively,administered by stomach 12 weeks.At the same time,C57 BL/6 mice were used as the normal control group,and deionized water was administered for 12 weeks.Using biochemical analyzer to detect triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(low-density lipoprotein cholesterol,LDL-C)content;HE staining to observe aortic vascular morphology,and measurement of aortic intima thickness(IT),medial thickness(MT),vascular lumen area(LA),atheroma area(PA)intima-media thickness ratio(IT/MT),lumen stenosis rate(PA/LA);interleukin-6(IL-6)and mononuclear serum,and expression of inflammatory cytokines by monocyte chemoattractant protein-1(MCP-1)were measured by flow-through high-throughput multifactor detection.Results:Compared with the control group,serum TC,TG and LDL-C in the model group were significantly increased,HDL-C was significantly decreased,and PA,IT and IT/MT were significantly increased(P<0.01).Compared with the model group,Simiao Yongan decoction significantly reduced PA,IT and IT/MT(P<0.01)in ApoE~(-/-)mice,and significantly decreased IL-6 and MCP-1 expression(P<0.01,P<0.05),but had no significant effect on blood lipid levels.Conclusion:Simiao Yongan decoction can prevent atherosclerosis by inhibiting inflammatory response,independent of changes in blood lipid levels.
Objective:To study the intervention effect of Simiao Yongan decoction on atherosclerotic inflammatory response in ApoE knockout mice fed with high fat diet.Methods:ApoE~(-/-)mice were fed with high-fat diet for 14 weeks to prepare atherosclerosis model.After modeling,they were randomly divided into 5 groups:model group,positive drug group(atorvastatin,2.57 mg·kg~(-1)),Simiao Yongan decoction high dose group(34.9 g·kg~(-1)),Simiao Yongan decoction medium dose group(23.14 g·kg~(-1))and Simiao Yongan low dose group(11.61 g·kg~(-1)),respectively,administered by stomach 12 weeks.At the same time,C57 BL/6 mice were used as the normal control group,and deionized water was administered for 12 weeks.Using biochemical analyzer to detect triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(low-density lipoprotein cholesterol,LDL-C)content;HE staining to observe aortic vascular morphology,and measurement of aortic intima thickness(IT),medial thickness(MT),vascular lumen area(LA),atheroma area(PA)intima-media thickness ratio(IT/MT),lumen stenosis rate(PA/LA);interleukin-6(IL-6)and mononuclear serum,and expression of inflammatory cytokines by monocyte chemoattractant protein-1(MCP-1)were measured by flow-through high-throughput multifactor detection.Results:Compared with the control group,serum TC,TG and LDL-C in the model group were significantly increased,HDL-C was significantly decreased,and PA,IT and IT/MT were significantly increased(P<0.01).Compared with the model group,Simiao Yongan decoction significantly reduced PA,IT and IT/MT(P<0.01)in ApoE~(-/-)mice,and significantly decreased IL-6 and MCP-1 expression(P<0.01,P<0.05),but had no significant effect on blood lipid levels.Conclusion:Simiao Yongan decoction can prevent atherosclerosis by inhibiting inflammatory response,independent of changes in blood lipid levels.
引文
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[22] 刘征辉,徐石勇,赵琳琳,等.金银花中抗炎活性成分獐牙菜苷和獐牙菜苦苷定量方法的研究[J].中国现代中药,2016,18(3):321-325.
[23] SOTILLO D V R D,HADLEY M.Chlorogenic acid modifies plasma and liver concentrations of:cholesterol,triacylglycerol,and minerals in(fa/fa)Zucker rats[J].J Nutr Biochem,2002,13(12):717-726.
[2] LIBBY P,RIDKER P M,MASERI A.Inflammation and atherosclerosis[J].Circulation,2002,105(9):1135-1143.
[3] 王立茹.四妙勇安汤治疗冠状动脉粥样硬化性心脏病60例[J].陕西中医,2010,31(2):131-132.
[4] 吕贵德.马同长主任医师治疗血栓闭塞性脉管炎经验介绍[J].新中医,2005,37(9):10-11.
[5] 陈辉.中西医结合治疗下肢深静脉血栓形成30例[J].中国中医急症,2006,15(1):7.
[6] 吴圣贤,聂波,潘美香,等.解毒活血法对ApoE-/-小鼠动脉粥样硬化斑块PPAR-γ和炎性因子的影响[J].辽宁中医杂志,2014,41(5):1046-1048.
[7] 徐冰,聂波,徐颖,等.基于ApoE-/-小鼠动脉粥样硬化模型的四妙勇安汤活性部位配伍规律研究[J].辽宁中医杂志,2013,40(6):1250-1252.
[8] PLUMP A S,SMITH J D,HAYEK T,et al.Severe hypercholesterolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells[J].Cell,1992,71(2):343-353.
[9] SORRENTINO S,LANDEMESSER U.Nonlipid-lowering effects of statins[J].Curr Treat Options Cardiovasc Med,2005,7(6):459-466.
[10] ZHOU Q,LIAO J K.Pleiotropic Effects of Statins:Basic Research and Clinical Perspectives[J].Circ J,2010,74(5):818-826
[11] ZADELAAR S,KLEEM ANNl R,VERSCHUREN L,et al.Mouse models for atherosclerosis and pharmaceutical modifiers[J].Arterioscl Throm Vas,2007,27(8):1706-1721.
[12] SPARROW C P,BURTON C A,HEMANDEZ M,et al.Simvastatin has anti-inflammatory and anti-atherosclerotic activities independent of plasma cholesterol-lowering[J].Atherosclerrosis,2001,151(1):188.
[13] YANG L,HENG X,GUO R,et al.Atorvastatin Inhibits the 5-Lipoxygenase Pathway and Expression of CCL3 to Alleviate Atherosclerotic Lesions in Atherosclerotic ApoE Knockout Mice[J].J Cardiovasc Pharm,2013,62(2):205-211.
[14] TZOULAKI I,MURRAY M G D,LEE P A J,et al.C-reactive protein,interleukin-6,and soluble adhesion molecules as predictors of progressive peripheral atherosclerosis in the general population:Edinburgh Artery Study[J].Acc Curr J Rev,2005,14(12):16.
[15] 杨月仙,刘波,张茂林.单核细胞趋化蛋白-1与动脉粥样硬化及脑梗死的关系[J].中国煤炭工业医学杂志,2011,14(3):462-465.
[16] KIZILAY MANCINI ?,LORA M,SHUM-TIM D,et al.A Proinflammatory Secretome Mediates the Impaired Immunopotency of Human Mesenchymal Stromal Cells in Elderly Patients With Atherosclerosis[J].Stem Cell Transl Med,2017,6(4):1132-1140.
[17] HAMLATKHENNAF N,NEGGAZI S,AYARI H,et al.Inflammation in the perivascular adipose tissue and atherosclerosis.[J].C R Biol,2017,340(3):156-163.
[18] BIANCONI V,SAHEBKAR A,ATKIN S L,et al.The regulation and importance of monocyte chemoattractant protein-1[J].Cerr Opin Hematol,2018,14(4):44-51.
[19] CASTI?EIRAS LANDEIR MI,RODI?O JANEIRO BK,PARADELADOBARRO B,et al.Change of concept about the regulation of angiotensin II-induced monocyte chemoattractant protein-1 production in human endothelial cells[J].Vasc Pharmacol,2016,80:20-34.
[20] ADAR S D,D′SOUZA J,MENDELSOHNVICTOR K,et al.Markers of inflammation and coagulation after long-term exposure to coarse particulate matter:a cross-sectional analysis from the multi-ethnic study of atherosclerosis[J].Environ Health Persp,2015,123(6):541-548.
[21] CLAHSEN T,SCHAPER F.Interleukin-6 acts in the fashion of a classical chemokine on monocytic cells by inducing integrin activation,cell adhesion,actin polymerization,chemotaxis,and transmigration[J].J Leukocyte Biol,2008,84(6):1521-1529.
[22] 刘征辉,徐石勇,赵琳琳,等.金银花中抗炎活性成分獐牙菜苷和獐牙菜苦苷定量方法的研究[J].中国现代中药,2016,18(3):321-325.
[23] SOTILLO D V R D,HADLEY M.Chlorogenic acid modifies plasma and liver concentrations of:cholesterol,triacylglycerol,and minerals in(fa/fa)Zucker rats[J].J Nutr Biochem,2002,13(12):717-726.