新活络效灵丹对ApoE基因敲除早期动脉粥样硬化小鼠Cav-1、SR-BI的影响
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摘要
目的:观察新活络效灵丹对ApoE基因敲除小鼠早期动脉粥样硬化(atherosclerosis,AS)斑块面积及对Cav-1、SR-BI表达的影响,探讨新活络效灵丹抗早期AS斑块的可能机制。方法:采用高脂饲料喂养ApoE基因敲除小鼠建立AS模型,将45只小鼠随机分为西药组、模型组、新活络效灵丹组,另将10只C57BL/6J小鼠设为空白组,连续给药13周后测量计算各组斑块面积及斑块/管腔面积比值(PA/ALA),检测各组主动脉Cav-1 mRNA、SR-BI mRNA表达水平。结果:与模型组比较,西药组斑块面积显著缩小,PA/ALA显著降低;中药组与模型组比较,斑块面积、PA/ALA均有显著改变;模型组小鼠主动脉组织中Cav-1 mRNA、SR-BI mRNA表达水平低于空白组;西药组表达水平高于模型组,中药组表达水平明显高于模型组,中西药组表达水平比较差异无统计学意义。结论:新活络效灵丹具有预防AS斑块形成的作用,是提高AS小鼠主动脉组织Cav-1 mRNA、SR-BI mRNA表达机制之一。
To investigate the effect of New Huoluo Xiaoling Dan( NHLXLD) on the area of early-stage atherosclerotic lesions and expression of Cav-1 and SR-B in ApoE Knock-out mice. To explore the mechanism of NHLXLD's reduction of early atherosclerotic plaque formation. Methods: AS model was induced by high fat diet in ApoE knock-out mice. A total of forty five ApoE knock-out mice were equally randomized into 3 groups,including control group( CG),western medicine group( WMG) and traditional Chinese medicine group( TCMG). Ten C57 BL/6 J mice were also treated as blank group( BG). During modeling,CG took normal saline 0. 3 ml daily; TCMG was treated with NHLXLD9. 67 g/kg. d,and WMG was administered avarstatin 3. 33 mg/kg. d. The treatment lasted for 13 weeks with gastric lavage once everyday. Then the aortic roots plaque area and its percentage of total surface area of the aortic arch was measured by software IPP5. 0 after HE staining. The real-time reverse transcription PCR( RT-PCR) was employed to determine the mRNA levels of SR-BI and Cav-1 in each group. Results: No changes were observed in blank group;whereas obvious atheroslerotic plaques were developed in control group. The plaque area and its ratio with aortic cavity area were decreased significantly in both WMG and TCMG,but no significant difference was observed between WMG and TCMG. The expressions level of SR-BⅠ mRNA and Cav-1 mRNA in control group were significantly lower than those in blank group. Elevated express of SR-BⅠmRNA and Cav-1 mRNA were observed in WMG,compared with control group;and the same results were obtained in TCMG with,TCMG demonstrated higher SR-B Ⅰ mRNA level,and lower Cav-1 mRNA level than WMG,which is not statistically significant. Conclusions: NHLXLD increased the inpression of SR-BⅠmRNA and Cav-1 mRNA in mice,thus enhance cholesterol efflux,which was proposed to be one of the mechanism of early-stage atherosclerotic plaque development.
To investigate the effect of New Huoluo Xiaoling Dan( NHLXLD) on the area of early-stage atherosclerotic lesions and expression of Cav-1 and SR-B in ApoE Knock-out mice. To explore the mechanism of NHLXLD's reduction of early atherosclerotic plaque formation. Methods: AS model was induced by high fat diet in ApoE knock-out mice. A total of forty five ApoE knock-out mice were equally randomized into 3 groups,including control group( CG),western medicine group( WMG) and traditional Chinese medicine group( TCMG). Ten C57 BL/6 J mice were also treated as blank group( BG). During modeling,CG took normal saline 0. 3 ml daily; TCMG was treated with NHLXLD9. 67 g/kg. d,and WMG was administered avarstatin 3. 33 mg/kg. d. The treatment lasted for 13 weeks with gastric lavage once everyday. Then the aortic roots plaque area and its percentage of total surface area of the aortic arch was measured by software IPP5. 0 after HE staining. The real-time reverse transcription PCR( RT-PCR) was employed to determine the mRNA levels of SR-BI and Cav-1 in each group. Results: No changes were observed in blank group;whereas obvious atheroslerotic plaques were developed in control group. The plaque area and its ratio with aortic cavity area were decreased significantly in both WMG and TCMG,but no significant difference was observed between WMG and TCMG. The expressions level of SR-BⅠ mRNA and Cav-1 mRNA in control group were significantly lower than those in blank group. Elevated express of SR-BⅠmRNA and Cav-1 mRNA were observed in WMG,compared with control group;and the same results were obtained in TCMG with,TCMG demonstrated higher SR-B Ⅰ mRNA level,and lower Cav-1 mRNA level than WMG,which is not statistically significant. Conclusions: NHLXLD increased the inpression of SR-BⅠmRNA and Cav-1 mRNA in mice,thus enhance cholesterol efflux,which was proposed to be one of the mechanism of early-stage atherosclerotic plaque development.
引文
[1]LIAO DF,TANG CK.Cholesterol reverse transport from basic to clinical[J].Beijing:Science and Technology Press(Chinese),2009,67.
[2]OHASHI R,MU H,WANG X,et al.Reverse cholesterol transport and efflux in atherosclerosis[J].Q J Med,2005,98(12):845-856.
[3]房玉涛,刘桂芳,张云.新活络效灵丹对Apo E基因敲除早期动脉粥样硬化小鼠TGF-β的影响[J].世界中医药,2016,11(10):2099.
[4]邓茹月.加减活络效灵丹对Apo E小鼠动脉粥样硬化的早期干预作用[J].北京中医药大学,2014:48-50.
[5]SUZUKI H,KURIHARA Y,TAKEYA M,et al.A role for macrophage scavenger receptors in atherosclerosis and susceptibility to infection[J].Nature,1997,386:292-296.
[6]JOHNSON J L,FRITSCHE-DANIELSON R,BEHRENDT M,et al.Effect of broad-spectrum matrix metalloproteinase inhibition on atherosclerotic plaque stability[J].Cardiovascular research,2006,71(3):586-595.
[7]张运.关于动脉粥样硬化的新治疗靶点[J].医学研究杂志,2011,40(6):1-3.
[8]ARGO CK,LORIA P,CALDWELL SH,et al.Statins in liver disease:amolehill,an iceberg,or neither[J].Hepatology,2008,48(2):662-669.
[9]USSO MW,SCOBEY M,BONKOVSKY HL.Drug-induced liver injury associated with statins[J].Semin Liver Dis,2009,29(4):412-422.
[10]ACEDO AF,TAYLOR FC,CASAS JP,et al.Unintended effects of statins from observational studies in the general population:Systematic review and meta-analysis[J].BMC medicine,2014,12(1):51.
[11]ANGPAISAN W,BRAYNE C.Alzheimers Society Vascular Dementia Systematic Review Group.Systematic review of statins for the prevention of vascular dementia or dementia[J].Geriatr Gerontol Int,2010,10(2):199-208.
[12]陈清杰.他汀类药物不良反应的研究进展[J].心血管病学进展,2013,34(2):279-283.
[13]MARINA C,RADER DJ.Macrophage reverse cholesterol transport-key to the regression of atherosclerosis[J].Circulation,2006,113:2548-2555.
[14]LI XA,EVERSON WV,SMART EJ.Caveolae,lipid rafts,and vascular disease[J].Trends Cardiovasc Med,2005,15(3):92-96.
[15]何清,王长谦.小凹蛋白1在动脉粥样硬化中的作用[J].中国动脉硬化杂志,2006,14(6):547-549.
[16]KIM S,LEE Y,SEO JE,et al.Caveolin-1 increases basal and TGF-beta1-induced expression of typeⅠprocollagen through PI-3 kinase/Akt/m TOR pathway in human dermal fibroblasts[J].Cell Signal,2008,20(7):1313-1319.
[17]DISTLER JH,HIRTH A,KUROWSKA-STOLARSK M,et al.Angiogenie and angiostatic faetors in the molecular control of angiogenesis[J].Q J Nucl Med,2003,47(3):149-161.
[18]FEI HAN,HONG-GUANG ZHU.Cav-1 Regulating the Invasion and Expression of Matrix Met alloproteinase(MMPs)in Pancreatic Carcinoma Cells[J].Journal of Surgical Research,2010,159(1):443–450.
[19]THMOS S,OVERDEVEST JB,NITZ MD,et al.Src and caveolin-1 reciprocally regulate metastasis via a common downstream signaling pathway in bladder cancer[J].Cancer Research,2011,71(3):101-109.
[20]WOOD WG,IGBAVBOA U,MULLER WE,et al.Cholesterol asymmetry in synaptic plasma membranes[J].J Neurochemistry,2011,116(5):684-689.
[21]廖端芳,覃丽.小凹及小凹蛋白-1:胆固醇逆向转运与炎症应答的共同分子平台[J].中国动脉硬化杂志,2012,20(5):385-392.
[22]LUO DX,CAO DL,XIONG Y,et al.A novel model of cholesterol efflux from lipid-loaded cells[J].Circulation,2009,119(8):135-145.
[23]ACTON S,RIGOTTI A,LANDSCHULZ KT,et al.Identification of scavenger receptor SR-Blas a high density Lipoprotein receptor[J].Scince,1996,271(5248):518-520.
[24]WEHINGER A,TANCEVSKI I,SEILER R,et al.Influence of aspirin on SR-BI expression in human carotid plaques[J].Atherosclerosis,2009,206:234-238.
[25]HONG SC,ZHAO SP,WU ZH.Effect of probucol on HDL metabolism and class B type I scavenger receptor(SR-BI)expression in the liver of hypercholesterolemic rabbits[J].Int J Cardiol,2007,115:29-35.
[26]HOEKSTRA M,VAN BERKEL TJ,VAN ECK M.Scavenger receptor BI:a multi-purpose player in cholesterol and steroidmetabolism[J].World J Gastroenterol,2010,16(47):5916-5924.
[27]陈可冀.活血化瘀方药降低心血管风险的可能性探索[J].中国中西医结合杂志,2008,28(5):389.
[28]邓茹月,房玉涛.活络效灵丹治疗心血管疾病的研究进展[J].贵阳中医学院学报,2014,36(3)3:142-144.
[29]徐英姿,韩向东.关于活络效灵丹的浅析[J].四川中医,2014,32(9):28.
[30]常允平,韩英梅,张俊艳.乳香的化学成分和药理活性研究进展[J].现代药物与临床,2012,27(1):52-59.
[31]HANUS L,REZANKA T,DEMBITSKV,et al.Myrrh-Commiphora[J].Chemistry,2005,149(1):23.
[32]朴春花.三七的药理作用研究进展概述[J].中国医药指南,2011,13(9):209-210.
[33]杨志霞,林谦,马利.丹参对心血管疾病药理作用的文献研究[J].世界中西医结合杂志,2012,7(2):93-97.
[2]OHASHI R,MU H,WANG X,et al.Reverse cholesterol transport and efflux in atherosclerosis[J].Q J Med,2005,98(12):845-856.
[3]房玉涛,刘桂芳,张云.新活络效灵丹对Apo E基因敲除早期动脉粥样硬化小鼠TGF-β的影响[J].世界中医药,2016,11(10):2099.
[4]邓茹月.加减活络效灵丹对Apo E小鼠动脉粥样硬化的早期干预作用[J].北京中医药大学,2014:48-50.
[5]SUZUKI H,KURIHARA Y,TAKEYA M,et al.A role for macrophage scavenger receptors in atherosclerosis and susceptibility to infection[J].Nature,1997,386:292-296.
[6]JOHNSON J L,FRITSCHE-DANIELSON R,BEHRENDT M,et al.Effect of broad-spectrum matrix metalloproteinase inhibition on atherosclerotic plaque stability[J].Cardiovascular research,2006,71(3):586-595.
[7]张运.关于动脉粥样硬化的新治疗靶点[J].医学研究杂志,2011,40(6):1-3.
[8]ARGO CK,LORIA P,CALDWELL SH,et al.Statins in liver disease:amolehill,an iceberg,or neither[J].Hepatology,2008,48(2):662-669.
[9]USSO MW,SCOBEY M,BONKOVSKY HL.Drug-induced liver injury associated with statins[J].Semin Liver Dis,2009,29(4):412-422.
[10]ACEDO AF,TAYLOR FC,CASAS JP,et al.Unintended effects of statins from observational studies in the general population:Systematic review and meta-analysis[J].BMC medicine,2014,12(1):51.
[11]ANGPAISAN W,BRAYNE C.Alzheimers Society Vascular Dementia Systematic Review Group.Systematic review of statins for the prevention of vascular dementia or dementia[J].Geriatr Gerontol Int,2010,10(2):199-208.
[12]陈清杰.他汀类药物不良反应的研究进展[J].心血管病学进展,2013,34(2):279-283.
[13]MARINA C,RADER DJ.Macrophage reverse cholesterol transport-key to the regression of atherosclerosis[J].Circulation,2006,113:2548-2555.
[14]LI XA,EVERSON WV,SMART EJ.Caveolae,lipid rafts,and vascular disease[J].Trends Cardiovasc Med,2005,15(3):92-96.
[15]何清,王长谦.小凹蛋白1在动脉粥样硬化中的作用[J].中国动脉硬化杂志,2006,14(6):547-549.
[16]KIM S,LEE Y,SEO JE,et al.Caveolin-1 increases basal and TGF-beta1-induced expression of typeⅠprocollagen through PI-3 kinase/Akt/m TOR pathway in human dermal fibroblasts[J].Cell Signal,2008,20(7):1313-1319.
[17]DISTLER JH,HIRTH A,KUROWSKA-STOLARSK M,et al.Angiogenie and angiostatic faetors in the molecular control of angiogenesis[J].Q J Nucl Med,2003,47(3):149-161.
[18]FEI HAN,HONG-GUANG ZHU.Cav-1 Regulating the Invasion and Expression of Matrix Met alloproteinase(MMPs)in Pancreatic Carcinoma Cells[J].Journal of Surgical Research,2010,159(1):443–450.
[19]THMOS S,OVERDEVEST JB,NITZ MD,et al.Src and caveolin-1 reciprocally regulate metastasis via a common downstream signaling pathway in bladder cancer[J].Cancer Research,2011,71(3):101-109.
[20]WOOD WG,IGBAVBOA U,MULLER WE,et al.Cholesterol asymmetry in synaptic plasma membranes[J].J Neurochemistry,2011,116(5):684-689.
[21]廖端芳,覃丽.小凹及小凹蛋白-1:胆固醇逆向转运与炎症应答的共同分子平台[J].中国动脉硬化杂志,2012,20(5):385-392.
[22]LUO DX,CAO DL,XIONG Y,et al.A novel model of cholesterol efflux from lipid-loaded cells[J].Circulation,2009,119(8):135-145.
[23]ACTON S,RIGOTTI A,LANDSCHULZ KT,et al.Identification of scavenger receptor SR-Blas a high density Lipoprotein receptor[J].Scince,1996,271(5248):518-520.
[24]WEHINGER A,TANCEVSKI I,SEILER R,et al.Influence of aspirin on SR-BI expression in human carotid plaques[J].Atherosclerosis,2009,206:234-238.
[25]HONG SC,ZHAO SP,WU ZH.Effect of probucol on HDL metabolism and class B type I scavenger receptor(SR-BI)expression in the liver of hypercholesterolemic rabbits[J].Int J Cardiol,2007,115:29-35.
[26]HOEKSTRA M,VAN BERKEL TJ,VAN ECK M.Scavenger receptor BI:a multi-purpose player in cholesterol and steroidmetabolism[J].World J Gastroenterol,2010,16(47):5916-5924.
[27]陈可冀.活血化瘀方药降低心血管风险的可能性探索[J].中国中西医结合杂志,2008,28(5):389.
[28]邓茹月,房玉涛.活络效灵丹治疗心血管疾病的研究进展[J].贵阳中医学院学报,2014,36(3)3:142-144.
[29]徐英姿,韩向东.关于活络效灵丹的浅析[J].四川中医,2014,32(9):28.
[30]常允平,韩英梅,张俊艳.乳香的化学成分和药理活性研究进展[J].现代药物与临床,2012,27(1):52-59.
[31]HANUS L,REZANKA T,DEMBITSKV,et al.Myrrh-Commiphora[J].Chemistry,2005,149(1):23.
[32]朴春花.三七的药理作用研究进展概述[J].中国医药指南,2011,13(9):209-210.
[33]杨志霞,林谦,马利.丹参对心血管疾病药理作用的文献研究[J].世界中西医结合杂志,2012,7(2):93-97.