丹皮酚减缓动脉粥样硬化病变的形成及发展涉及脂质调节,抗炎和抗氧化活性（英文）
|
摘要
丹皮酚是从中国传统中药材牡丹皮根部所提取的活性成分,具有抗炎和保护心血管等多种生物活性。在本次实验中,丹皮酚抗动脉粥样硬化的效应贯穿于Apo E基因缺陷小鼠动脉粥样硬化疾病发展的各个时期。将Apo E基因缺陷小鼠随机分成五组,并分别给予高胆固醇(HCD)喂养5、15、25周,然后分别灌胃给丹皮酚或阿托伐他汀6周时间。采用ELISA法检测血清中脂质和细胞因子水平,蛋白质印迹法检测蛋白表达,采用H&E和油红染色法对胸主动脉病变区域进行评估。结果显示,在动脉粥样硬化疾病过程的各个阶段,丹皮酚均能显着降低ApoE~(–/–)小鼠的体重、血脂、总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白(LDL-C)水平。丹皮酚还可降低血清中的炎症因子、如肿瘤坏死因子-α(TNF-α)、白介素(IL)-6、氧化型低密度脂蛋白胆固醇(ox-LDL-C)水平。在丹皮酚组中超氧化物歧化酶(SOD)与HCD组相比显著增加而MDA(丙二醛)水平显著降低(P<0.01)丹皮酚显著减弱富含脂质斑块的厚度,下调小鼠主动脉中血管细胞粘附因子(VCAM-1)和金属基质蛋白酶(MMP-9)的表达,这表明丹皮酚可抑制斑块形成并稳定斑块。丹皮酚在Apo E基因缺陷小鼠动脉粥样硬化疾病发展的各个时期发挥了调节脂质、抗炎和抗氧化的效应,显示出其具有抗血脂异常和抗动脉粥样硬化的作用。
As an active component extracted from Chinese herb moutan cortex and roots of paeonia lactiflora pallas, paeonol possesses many biological activities, including anti-inflammatory and vascular protection activities. In this study, athero-protective effects of paeonol were investigated in apoliprotein E deficient(Apo E~(–/–)) mice through the entire course of atherosclerotic development. Apo E~(–/–) mice were divided into five groups and fed a high-cholesterol diet(HCD) for 5, 15 and 25 weeks. Then they were fed either paeonol or atorvastatin for 6 weeks. The methods, such as ELISA for serum lipid and cytokine analyses, Western blotting for protein expressions, and H&E and oil-red o-staining method, were used for evaluation of thoracic aorta lesion area. The results showed that paeonol could significantly reduce body weight, blood lipid, total cholesterol(TC), triglyceride(TG) and low density lipoprotein(LDL-C) in Apo E~(–/–) mice at all stages of the atherosclerosis process. Paeonol also reduced the levels of anti-inflammation factors, such as tumor necrosis factor-α(TNF-α), interleukin(IL)-6, oxidized LDL cholesterol(ox-LDL), in serum. In paeonol groups, SOD was significantly increased, whereas MDA was decreased compared with the HCD group(P<0.01). Paeonol markedly attenuated the thickness of the lipid-rich plaque and down-regulated the expressions of VCAM-1 and MMP-9 in aorta of mice, suggesting that paeonol could inhibit formation of plaque and stabilize plaques. Taken together, paeonol appeared to have anti-dyslipidemia and anti-atherosclerotic effects by lipid regulation, and it could inhibit the effects of inflammation and oxidative stress on HCD-fed Apo E~(–/–) mice through the entire course of atherosclerotic development.
As an active component extracted from Chinese herb moutan cortex and roots of paeonia lactiflora pallas, paeonol possesses many biological activities, including anti-inflammatory and vascular protection activities. In this study, athero-protective effects of paeonol were investigated in apoliprotein E deficient(Apo E~(–/–)) mice through the entire course of atherosclerotic development. Apo E~(–/–) mice were divided into five groups and fed a high-cholesterol diet(HCD) for 5, 15 and 25 weeks. Then they were fed either paeonol or atorvastatin for 6 weeks. The methods, such as ELISA for serum lipid and cytokine analyses, Western blotting for protein expressions, and H&E and oil-red o-staining method, were used for evaluation of thoracic aorta lesion area. The results showed that paeonol could significantly reduce body weight, blood lipid, total cholesterol(TC), triglyceride(TG) and low density lipoprotein(LDL-C) in Apo E~(–/–) mice at all stages of the atherosclerosis process. Paeonol also reduced the levels of anti-inflammation factors, such as tumor necrosis factor-α(TNF-α), interleukin(IL)-6, oxidized LDL cholesterol(ox-LDL), in serum. In paeonol groups, SOD was significantly increased, whereas MDA was decreased compared with the HCD group(P<0.01). Paeonol markedly attenuated the thickness of the lipid-rich plaque and down-regulated the expressions of VCAM-1 and MMP-9 in aorta of mice, suggesting that paeonol could inhibit formation of plaque and stabilize plaques. Taken together, paeonol appeared to have anti-dyslipidemia and anti-atherosclerotic effects by lipid regulation, and it could inhibit the effects of inflammation and oxidative stress on HCD-fed Apo E~(–/–) mice through the entire course of atherosclerotic development.
引文
[1]Libby,P.;Aikawa,M.Stabilization of atherosclerotic plaque:new mechanisms and clinical targets.Nat.Med.2002,8,1257-1262.
[2]Lusis,A.J.Atherosclerosis.Nature.2000,407,233-241.
[3]Kinkade,K.;Streeter,J.;Miller,F.J.Inhibition of NADPH Oxidase by Apocynin Attenuates Progression of Atherosclerosis.Int.J.Mol.Sci.2013,14,17017-17028.
[4]Chobanian,A.V.Single risk factor intervention may be inadequate to inhibit atherosclerosis progression when hypertension and hypercholesterolemia coexist.Hypertension.1991,18,130-131.
[5]Stoll,G.;Bendszus,M.Inflammation and atherosclerosis:novel insights into plaque formation and destabilization.Stroke.2006,37,1923-1932.
[6]Yung,L.M.;Leung,F.P.;Yao,X.;Chen,Z.Y.;Huang,Y.Reactive oxygen species in vascular wall.Cardiovasc.Hematol.Disord.Drug Targets.2006,6,1-19.
[7]Bao,M.H;Zhang,Y.W.;Zhou,H.H.Paeonol suppresses oxidized low-density lipoprotein induced endothelial cell apoptosis via activation of LOX-1/p38MAPK/NF-κBpathway.J.Ethnopharmacol.2013,146,543-551.
[8]Shi,L.;Fan,P.S.;Fang,J.X.;Han,Z.X.Inhibitory effects of paeonol on experimental atherosclerosis and platelet aggregation of rabbit.Acta Pharmacol.Sin.1998,9,555-558.
[9]Lau,C.H.;Chan,C.M.;Chan,Y.W.;Lau,K.M.;Lau,T.W.;Lam,F.C.;Law,W.T.;Che,C.T.;Leung,P.C.;Fung,K.P.;Ho,Y.Y.;Lau,C.B.Pharmacological investigations of the anti-diabetic effect of Cortex Moutan and its active component paeonol.Phytomedicine.2007,14,778-784.
[10]Sun,G.P.;Wang,H.;Xu,S.P.;Wu,Q.;Chen,Z.D.;Wei,W.Anti-tumor effects of paeonol in a Hep A-hepatoma bearing mouse model via induction of tumor cell apoptosis and stimulation of IL-2 and TNF-αproduction.Eur.J.Pharmacol.2008,584,246-252.
[11]Pan,L.L.;Dai,M.Paeonol from Paeonia suffruticosa prevents TNF-alpha-induced monocytic cell adhesion to rat aortic endothelial cells by suppression of VCAM-1expression.Phytomedicine Inter.J.Phytotherapy Phytopharmacology.2009,16,1027-1032.
[12]Liu,Y.R.;Chen,J.J.;Dai,M.Paeonol protects rat vascular endothelial cells from ox-LDL-induced injury in vitro via downregulating micro RNA-21 expression and TNF-αrelease.Acta Pharmacol.Sin.2014,35,483-488.
[13]Wu,H.F.;Su,X.F.;Dai,M.Paeonol Inhibits Overproliferation of VSMCs in ox-LDL-Injured co-cultured System by Suppression of ET-1 Expression.Lat.Am.J.Pharm.2015,34,1858-1865.
[14]Dai,M.;Zhi,X.M.;Peng,D.Y.;Liu,Q.Y.Inhibitory Effect of Paeonol on Experimental Atherosclerosis in Quails.China J.Chin.Mater.Med.1999,24,488-490.
[15]Li,H.K.;Dai,M.;Jia,W.Paeonol attenuates high-fatdiet-induced atherosclerosis in rabbits by anti-inflammatory activity.Planta Med.2009,75,7-11.
[16]Shan,L.M.;Zhang,J.C.;Zhao,Y.L.;Wang,H.Primary Mechanisms for Novel Compound Pivanampeta Against Atherosclerosis in Rat and Rabbit Model of Atherosclerosis.J.Chin.Pharm.Sci.2004,13,1.
[17]Getz,G.S.;Reardon,C.A.Animal models of atherosclerosis.Arterioscler.Thromb.Vasc.Biol.2012,32,1104-1115.
[18]Kuehn,C.;Tauchi,M.;Stumpf,C.;Daniel,C.;B?uerle,T.;Schwarz,M.;Kerek,F.;Steinkasserer,A.;Zinser,E.;Achenbach,S.;Dietel,B.Suppression of proatherogenic leukocyte interactions by MCS-18-Impact on advanced atherosclerosis in Apo E-deficient mice.Atherosclerosis.2016,245,101-110.
[19]Zhang,S.H.;Reddick,R.L.;Piedrahita,J.A.;Maeda,N.Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E.Science.1992,258,468-471.
[20]Nakashima,Y.;Plump,A.S.;Raines,E.W.;Breslow,J.L.;Ross,R.Apo E-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree.Arterioscler.Thromb.Vasc.Biol.1994,14,133-140.
[21]Nagarajan,S.;Burris,R.L.;Stewart,B.W.;Wilkerson,J.E.;Badger,T.M.Dietary soy protein isolate ameliorates atherosclerotic lesions in apolipoprotein E-deficient mice potentially by inhibiting monocyte chemoattractant protein-1 expression.J.Nutr.2008,138,332-337.
[22]Choi,S.H.;Chae,A.;Miller,E.;Ntanios,F.;De Maria,A.N.;Nissen,S.E.;Witztum,J.L.;Tsimikas,S.Relationship Between Biomarkers of Oxidized Low-Density Lipoprotein,Statin Therapy,Quantitative Coronary Angiography,and Atheroma Volume:Observations From the REVERSAL(Reversal of Atherosclerosis with Aggressive Lipid Lowering)Study.J.Am.Coll.Cardiol.2008,52,24-32.
[23]Meir,E.Leitersdorf,Atherosclerosis in the apolipoproteinE-deficient mouse:a decade of progress.Arterioscler.Thromb.Vasc.Biol.2004,24,1006-1014.
[24]Wang,Z.H.;Li,Y.F.;Guo,Y.Q.β3-Adrenoceptor activation attenuates atherosclerotic plaque formation in Apo E-/-mice through lowering blood lipids and glucose.Acta Pharmacol.Sin.2013,34,1156-1163.
[25]Tabet,F.;Rye,K.A.High-density lipoproteins,inflammation and oxidative stress.Clin.Sci.(Lond).2009,116,87-98.
[26]Mcneil,C.J.;Beattie,J.H.;Gordon,M.J.;Pirie,L.;Duthie,S.J.Nutritional B vitamin deficiency disrupts lipid metabolism causing accumulation of proatherogenic lipoproteins in the aorta adventitia of Apo E null mice.Mol.Nutr.Food Res.2012,56,1122-1130.
[27]Ross,R.Atherosclerosis-An Inflammatory Disease.N.Engl.J.Med.1999,340,115-126.
[28]Weber,C.;Noels,H.Atherosclerosis:current pathogenesis and therapeutic options.Nat.Med.2011,17,1410-1422.
[29]Libby,P.Inflammation in atherosclerosis.Nature.2002,420,868-874.
[30]Pirillo,A.;Norata,G.D.;Catapano,A.L.LOX-1,ox-LDL,and atherosclerosis.Med.Inf.2013,5,519-530.
[31]Gotsman,I.;Stabholz,A.;Planer,D.;Pugatsch,T.;Lapidus,L.;Novikov,Y.;Masrawa,S.;Soskolne,A.;Lotan,C.Serum cytokine tumor necrosis factoralpha and interleukin-6 associated with the severity of coronary artery disease:indicators of an active inflammatory burden?Isr.Med.Assoc.J.2008,10,494-498.
[32]Victor,V.M.;Apostolova,N.;Herance,R.;HernandezMijares,A.;Rocha,M.Oxidative stress and mitochondrial dysfunction in atherosclerosis:mitochondria-targeted antioxidants as potential therapy.Curr Med.Chem.2009,16,4654-667.
[33]Verreth,W.;De-Keyzer,D.P.;Geeraert,B.;Mertens,A.;Herregods,M.C.;Smith,G.;Desjardins,F.;Balligand,J.L.;Holvoet,P.Rosuvastatin restores superoxide dismutase expression and inhibits accumulation of oxidized LDLin the aortic arch of obese dyslipidemic mice.Br.J.Pharmacol.2007,151,347-355.
[34]Cybulsky,M.I.;Iiyama,K.;Li,H.;Zhu,S.;Chen,M.;Iiyama,M.;Davis,V.;Gutierrez-Ramos,J.C.;Connelly,P.W.;Milstone,D.S.A major role for VCAM-1,but not ICAM-1,in early atherosclerosis.J.Clin.Invest.2001,107,1255-1262.
[35]Johnson,J.L.;Devel,L.;Czarny,B.;George,S.J.;Jackson,C.L.;Rogakos,V.;Beau,F.;Yiotakis,A.;Newby,A.C.;Dive,V.A selective matrix metalloproteinase-12inhibitor retards atherosclerotic plaque development in apolipoprotein E-knockout mice.Arterioscler.Thromb.Vasc.Biol.2011,31,528-535.
[36]Halade,G.V.;Jin,Y.F.;Lindsey,M.L.Matrix metalloproteinase(MMP)-9:a proximal biomarker for cardiac remodeling and a distal biomarker for inflammation.Pharmacol.Ther.2013,139,32-40.
[2]Lusis,A.J.Atherosclerosis.Nature.2000,407,233-241.
[3]Kinkade,K.;Streeter,J.;Miller,F.J.Inhibition of NADPH Oxidase by Apocynin Attenuates Progression of Atherosclerosis.Int.J.Mol.Sci.2013,14,17017-17028.
[4]Chobanian,A.V.Single risk factor intervention may be inadequate to inhibit atherosclerosis progression when hypertension and hypercholesterolemia coexist.Hypertension.1991,18,130-131.
[5]Stoll,G.;Bendszus,M.Inflammation and atherosclerosis:novel insights into plaque formation and destabilization.Stroke.2006,37,1923-1932.
[6]Yung,L.M.;Leung,F.P.;Yao,X.;Chen,Z.Y.;Huang,Y.Reactive oxygen species in vascular wall.Cardiovasc.Hematol.Disord.Drug Targets.2006,6,1-19.
[7]Bao,M.H;Zhang,Y.W.;Zhou,H.H.Paeonol suppresses oxidized low-density lipoprotein induced endothelial cell apoptosis via activation of LOX-1/p38MAPK/NF-κBpathway.J.Ethnopharmacol.2013,146,543-551.
[8]Shi,L.;Fan,P.S.;Fang,J.X.;Han,Z.X.Inhibitory effects of paeonol on experimental atherosclerosis and platelet aggregation of rabbit.Acta Pharmacol.Sin.1998,9,555-558.
[9]Lau,C.H.;Chan,C.M.;Chan,Y.W.;Lau,K.M.;Lau,T.W.;Lam,F.C.;Law,W.T.;Che,C.T.;Leung,P.C.;Fung,K.P.;Ho,Y.Y.;Lau,C.B.Pharmacological investigations of the anti-diabetic effect of Cortex Moutan and its active component paeonol.Phytomedicine.2007,14,778-784.
[10]Sun,G.P.;Wang,H.;Xu,S.P.;Wu,Q.;Chen,Z.D.;Wei,W.Anti-tumor effects of paeonol in a Hep A-hepatoma bearing mouse model via induction of tumor cell apoptosis and stimulation of IL-2 and TNF-αproduction.Eur.J.Pharmacol.2008,584,246-252.
[11]Pan,L.L.;Dai,M.Paeonol from Paeonia suffruticosa prevents TNF-alpha-induced monocytic cell adhesion to rat aortic endothelial cells by suppression of VCAM-1expression.Phytomedicine Inter.J.Phytotherapy Phytopharmacology.2009,16,1027-1032.
[12]Liu,Y.R.;Chen,J.J.;Dai,M.Paeonol protects rat vascular endothelial cells from ox-LDL-induced injury in vitro via downregulating micro RNA-21 expression and TNF-αrelease.Acta Pharmacol.Sin.2014,35,483-488.
[13]Wu,H.F.;Su,X.F.;Dai,M.Paeonol Inhibits Overproliferation of VSMCs in ox-LDL-Injured co-cultured System by Suppression of ET-1 Expression.Lat.Am.J.Pharm.2015,34,1858-1865.
[14]Dai,M.;Zhi,X.M.;Peng,D.Y.;Liu,Q.Y.Inhibitory Effect of Paeonol on Experimental Atherosclerosis in Quails.China J.Chin.Mater.Med.1999,24,488-490.
[15]Li,H.K.;Dai,M.;Jia,W.Paeonol attenuates high-fatdiet-induced atherosclerosis in rabbits by anti-inflammatory activity.Planta Med.2009,75,7-11.
[16]Shan,L.M.;Zhang,J.C.;Zhao,Y.L.;Wang,H.Primary Mechanisms for Novel Compound Pivanampeta Against Atherosclerosis in Rat and Rabbit Model of Atherosclerosis.J.Chin.Pharm.Sci.2004,13,1.
[17]Getz,G.S.;Reardon,C.A.Animal models of atherosclerosis.Arterioscler.Thromb.Vasc.Biol.2012,32,1104-1115.
[18]Kuehn,C.;Tauchi,M.;Stumpf,C.;Daniel,C.;B?uerle,T.;Schwarz,M.;Kerek,F.;Steinkasserer,A.;Zinser,E.;Achenbach,S.;Dietel,B.Suppression of proatherogenic leukocyte interactions by MCS-18-Impact on advanced atherosclerosis in Apo E-deficient mice.Atherosclerosis.2016,245,101-110.
[19]Zhang,S.H.;Reddick,R.L.;Piedrahita,J.A.;Maeda,N.Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E.Science.1992,258,468-471.
[20]Nakashima,Y.;Plump,A.S.;Raines,E.W.;Breslow,J.L.;Ross,R.Apo E-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree.Arterioscler.Thromb.Vasc.Biol.1994,14,133-140.
[21]Nagarajan,S.;Burris,R.L.;Stewart,B.W.;Wilkerson,J.E.;Badger,T.M.Dietary soy protein isolate ameliorates atherosclerotic lesions in apolipoprotein E-deficient mice potentially by inhibiting monocyte chemoattractant protein-1 expression.J.Nutr.2008,138,332-337.
[22]Choi,S.H.;Chae,A.;Miller,E.;Ntanios,F.;De Maria,A.N.;Nissen,S.E.;Witztum,J.L.;Tsimikas,S.Relationship Between Biomarkers of Oxidized Low-Density Lipoprotein,Statin Therapy,Quantitative Coronary Angiography,and Atheroma Volume:Observations From the REVERSAL(Reversal of Atherosclerosis with Aggressive Lipid Lowering)Study.J.Am.Coll.Cardiol.2008,52,24-32.
[23]Meir,E.Leitersdorf,Atherosclerosis in the apolipoproteinE-deficient mouse:a decade of progress.Arterioscler.Thromb.Vasc.Biol.2004,24,1006-1014.
[24]Wang,Z.H.;Li,Y.F.;Guo,Y.Q.β3-Adrenoceptor activation attenuates atherosclerotic plaque formation in Apo E-/-mice through lowering blood lipids and glucose.Acta Pharmacol.Sin.2013,34,1156-1163.
[25]Tabet,F.;Rye,K.A.High-density lipoproteins,inflammation and oxidative stress.Clin.Sci.(Lond).2009,116,87-98.
[26]Mcneil,C.J.;Beattie,J.H.;Gordon,M.J.;Pirie,L.;Duthie,S.J.Nutritional B vitamin deficiency disrupts lipid metabolism causing accumulation of proatherogenic lipoproteins in the aorta adventitia of Apo E null mice.Mol.Nutr.Food Res.2012,56,1122-1130.
[27]Ross,R.Atherosclerosis-An Inflammatory Disease.N.Engl.J.Med.1999,340,115-126.
[28]Weber,C.;Noels,H.Atherosclerosis:current pathogenesis and therapeutic options.Nat.Med.2011,17,1410-1422.
[29]Libby,P.Inflammation in atherosclerosis.Nature.2002,420,868-874.
[30]Pirillo,A.;Norata,G.D.;Catapano,A.L.LOX-1,ox-LDL,and atherosclerosis.Med.Inf.2013,5,519-530.
[31]Gotsman,I.;Stabholz,A.;Planer,D.;Pugatsch,T.;Lapidus,L.;Novikov,Y.;Masrawa,S.;Soskolne,A.;Lotan,C.Serum cytokine tumor necrosis factoralpha and interleukin-6 associated with the severity of coronary artery disease:indicators of an active inflammatory burden?Isr.Med.Assoc.J.2008,10,494-498.
[32]Victor,V.M.;Apostolova,N.;Herance,R.;HernandezMijares,A.;Rocha,M.Oxidative stress and mitochondrial dysfunction in atherosclerosis:mitochondria-targeted antioxidants as potential therapy.Curr Med.Chem.2009,16,4654-667.
[33]Verreth,W.;De-Keyzer,D.P.;Geeraert,B.;Mertens,A.;Herregods,M.C.;Smith,G.;Desjardins,F.;Balligand,J.L.;Holvoet,P.Rosuvastatin restores superoxide dismutase expression and inhibits accumulation of oxidized LDLin the aortic arch of obese dyslipidemic mice.Br.J.Pharmacol.2007,151,347-355.
[34]Cybulsky,M.I.;Iiyama,K.;Li,H.;Zhu,S.;Chen,M.;Iiyama,M.;Davis,V.;Gutierrez-Ramos,J.C.;Connelly,P.W.;Milstone,D.S.A major role for VCAM-1,but not ICAM-1,in early atherosclerosis.J.Clin.Invest.2001,107,1255-1262.
[35]Johnson,J.L.;Devel,L.;Czarny,B.;George,S.J.;Jackson,C.L.;Rogakos,V.;Beau,F.;Yiotakis,A.;Newby,A.C.;Dive,V.A selective matrix metalloproteinase-12inhibitor retards atherosclerotic plaque development in apolipoprotein E-knockout mice.Arterioscler.Thromb.Vasc.Biol.2011,31,528-535.
[36]Halade,G.V.;Jin,Y.F.;Lindsey,M.L.Matrix metalloproteinase(MMP)-9:a proximal biomarker for cardiac remodeling and a distal biomarker for inflammation.Pharmacol.Ther.2013,139,32-40.