桃红四物汤干预外周血内皮祖细胞数量与功能增加的时间剂量效应
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摘要
背景:近期研究表明,桃红四物汤能减轻血管内皮细胞的损伤和维持其正常的分泌功能,而内皮祖细胞对内皮损伤后的修复有促进作用,因而设想桃红四物汤可能通过改善内皮祖细胞的功能活性及增加其数量来保护内皮功能。目的:观察桃红四物汤对人外周血内皮祖细胞功能活性和数量的影响。方法:从健康人外周血中分离培养内皮祖细胞,将内皮祖细胞分为对照组和桃红四物汤低、中、高质量浓度组,在体外培养24 h以观察量效关系,同时桃红四物汤高质量浓度组培养6,12,24,48 h以观察时效关系。倒置相差显微镜下计数内皮祖细胞数量,通过MTT比色法、黏附能力和Boyden小室观察内皮祖细胞的增殖、黏附和迁移能力。结果与结论:(1)桃红四物汤干预组内皮祖细胞增殖、黏附和迁移能力明显高于对照组,并与质量浓度呈一定的量效关系,高质量浓度组最为显著(P <0.01);(2)桃红四物汤高质量浓度组内皮祖细胞增殖、黏附和迁移能力的增加呈时间依赖关系,干预24 h时最为显著(P <0.01);(3)结果表明,桃红四物汤可增加内皮祖细胞数量并改善其功能,高质量浓度干预24 h时效果最佳。
BACKGROUND: Recent studies have shown that Taohong Siwu Decoction can alleviate the damage of vascular endothelial cells and maintain their normal secretory function, and endothelial progenitor cells can promote the repair of endothelial injury. Therefore, it is assumed that Taohong Siwu Decoction may protect endothelial function by improving the functional activity and increasing the number of endothelial progenitor cells. OBJECTIVE: To investigate whether Taohong Siwu Decoction can augment the number and functional activity of peripheral blood endothelial progenitor cells. METHODS: Endothelial progenitor cells were isolated from the peripheral blood of healthy subjects, and divided into control, low-, moderateand high-concentration Taohong Siwu Decoction groups. Cells were then cultured to observe the dose-effect relationship within 24 hours. Meanwhile, the high-concentration Taohong Siwu Decoction group was cultured for respective time points(6, 12, 24 and 48 hours) for observing the time-effect relationship. The number of endothelial progenitor cells was counted under inverted phase contrast microscope. Proliferation, adhesion and migration of endothelial progenitor cells were detected by MTT chromatometry, adhesion activity assay and modified Boyden chamber assay, respectively. RESULTS AND CONCLUSION:(1) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were significantly higher than those in the control group and showed a certain dose-effect relationship.(2) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were enhanced in a time-dependent manner, especially at 24 hours after intervention(P < 0.01). To conclude, the Taohong Siwu Decoction can increase the number of endothelial progenitor cells and promote cell functions. High-concentration Taohong Siwu Decoction exhibits the best interventional effect at 24 hours after intervention.
BACKGROUND: Recent studies have shown that Taohong Siwu Decoction can alleviate the damage of vascular endothelial cells and maintain their normal secretory function, and endothelial progenitor cells can promote the repair of endothelial injury. Therefore, it is assumed that Taohong Siwu Decoction may protect endothelial function by improving the functional activity and increasing the number of endothelial progenitor cells. OBJECTIVE: To investigate whether Taohong Siwu Decoction can augment the number and functional activity of peripheral blood endothelial progenitor cells. METHODS: Endothelial progenitor cells were isolated from the peripheral blood of healthy subjects, and divided into control, low-, moderateand high-concentration Taohong Siwu Decoction groups. Cells were then cultured to observe the dose-effect relationship within 24 hours. Meanwhile, the high-concentration Taohong Siwu Decoction group was cultured for respective time points(6, 12, 24 and 48 hours) for observing the time-effect relationship. The number of endothelial progenitor cells was counted under inverted phase contrast microscope. Proliferation, adhesion and migration of endothelial progenitor cells were detected by MTT chromatometry, adhesion activity assay and modified Boyden chamber assay, respectively. RESULTS AND CONCLUSION:(1) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were significantly higher than those in the control group and showed a certain dose-effect relationship.(2) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were enhanced in a time-dependent manner, especially at 24 hours after intervention(P < 0.01). To conclude, the Taohong Siwu Decoction can increase the number of endothelial progenitor cells and promote cell functions. High-concentration Taohong Siwu Decoction exhibits the best interventional effect at 24 hours after intervention.
引文
[1]邓中甲.方剂学[M].北京:中国中医药出版社,2008:240.
[2]韩岚,许钒,章小兵,等.桃红四物汤活血化瘀作用的实验研究[J].安徽中医学院学报,2007,26(1):36-37.
[3]孔增科,周海平,付正良.常用中药药理与临床应用[M].赤峰:内蒙古科学技术出版社,2005:257-286.
[4]胡德胜,郭伟星,于杰.心血管疾病活血化瘀论治[J].中西医结合心脑血管病杂志,2006,4(1):66-67.
[5]李鑫,李大勇,陈文娜,等.ASO血瘀证血清对血管内皮细胞损伤的影响及桃红四物汤的调节作用[J].中国中西医结合杂志,2015,35(11):1373-1377.
[6]张健,张宇,马贤德,等.光老化人真皮微血管内皮细胞模型的建立及桃红四物汤的保护作用[J].辽宁中药杂志,2018,45(6):1296-1299.
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[9]Yamahara K,Itoh H.Potential use of endothelial progenitor cells for regeneration of the vasculature.Ther Adv Cardiovasc Dis.2009;3(1):17-27.
[10]Kawamoto A,Asahara T.Role of progenitor endothelial cells in cardiovascular disease and upcoming therapies.Catheter Cardiovasc Interv.2007;70(4):477-484.
[11]Kalka C,Masuda H,Takahashi T,et al.Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization.Proc Natl Acad Sci U S A.2000;97(7):3422-3427.
[12]Tepper OM,Galiano RD,Capla JM,et al.Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation,adhesion,and incorporation into vascular structures.Circulation.2002;106(22):2781-2786.
[13]刘艳玲,李大勇,李雪晶.桃红四物汤调节动脉硬化闭塞症模型大鼠血液流动性的研究[J].中华中医药学刊,2014,32(4):761-763.
[14]张海峰,戴华卫,周瑞耀,等.酸性成纤维细胞生长因子对人脐带血内皮祖细胞凋亡的影响[J].中国动脉硬化杂志,2013,21(2):154-158.
[15]Peichev M,Naiyer AJ,Pereira D,et al.Expression of VEGFR-2 and AC133 by circulating human CD34(+)cells identifies a population of functional endothelial precursors.Blood.2000;95(3):952-958.
[16]王小斌,黄峻,朱莉,等.白藜三醇对人外周血内皮祖细胞功能的影响[J].江苏医学,2009,35(8):901-904.
[17]齐振熙,康靖东.桃红四物汤干预激素性股骨头缺血坏死组织局部微血管密度的变化[J].中国组织工程研究与临床康复,2009,13(11):2040-2043.
[18]龙翔,李娟,孙绍裘.桃红四物汤对兔皮片移植模型血管新生及MMP-2的影响[J].中医药导报,2016,22(22):14-17.
[19]池晓霞.桃红四物汤促进内皮细胞增殖及其机制初探[J].海峡药学,2010,22(1):39-41.
[20]刘竹青,尹登科,韩岚,等.桃红四物汤含药血清对过氧化氢损伤的人脐静脉内皮细胞的保护作用[J].中国中药杂,2013,38(3):402-406.
[21]Asahara T,Murohara T,Sullivan A,et al.Isolation of putative progenitor endothelial cells for angiogenesis.Science.1997;275(5302):964-967.
[22]Balbarini A,Barsotti MC,Di Stefano R,et al.Circulating endothelial progenitor cells characterization,function and relationship with cardiovascular risk factors.Curr Pharm Des.2007;13(16):1699-1713.
[23]Reyes M,Dudek A,Jahagirdar B,et al.Origin of endothelial progenitors in human postnatal bone marrow.J Clin Invest.2002;109(3):337-346.
[24]Werner N,Nickenig G.Influence of cardiovascular risk factors on endothelial progenitor cells:limitations for therapy.Arterioscler Thromb Vasc Biol.2006;26(2):257-266.
[25]Murayama T,Tepper OM,Silver M,et al.Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo.Exp Hematol.2002;30(8):967-972.
[26]Suzuki T,Nishida M,Futami S,et al.Neoendothelialization after peripheral blood stem cell transplantation in humans:a case report of a Tokaimura nuclear accident victim.Cardiovasc Res.2003;58(2):487-492.
[27]Vasa M,Fichtlscherer S,Aicher A,et al.Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease.Circ Res.2001;89(1):E1-7.
[28]Eizawa T,Ikeda U,Murakami Y,et al.Decrease in circulating endothelial progenitor cells in patients with stable coronary artery disease.Heart.2004;90(6):685-686.
[29]Loomans CJ,de Koning EJ,Staal FJ,et al.Endothelial progenitor cell dysfunction:a novel concept in the pathogenesis of vascular complications of type 1 diabetes.Diabetes.2004;53(1):195-199.
[30]Quyyumi AA.Circulating endothelial progenitor cells as novel biological determinants of vascular function and risk.Can JCardiol.2004;20 Suppl B:44B-48B.
[2]韩岚,许钒,章小兵,等.桃红四物汤活血化瘀作用的实验研究[J].安徽中医学院学报,2007,26(1):36-37.
[3]孔增科,周海平,付正良.常用中药药理与临床应用[M].赤峰:内蒙古科学技术出版社,2005:257-286.
[4]胡德胜,郭伟星,于杰.心血管疾病活血化瘀论治[J].中西医结合心脑血管病杂志,2006,4(1):66-67.
[5]李鑫,李大勇,陈文娜,等.ASO血瘀证血清对血管内皮细胞损伤的影响及桃红四物汤的调节作用[J].中国中西医结合杂志,2015,35(11):1373-1377.
[6]张健,张宇,马贤德,等.光老化人真皮微血管内皮细胞模型的建立及桃红四物汤的保护作用[J].辽宁中药杂志,2018,45(6):1296-1299.
[7]Hill JM,Zalos G,Halcox JP,et al.Circulating endothelial progenitor cells,vascular function,and cardiovascular risk.NEngl J Med.2003;348(7):593-600.
[8]Walter DH,Rittig K,Bahlmann FH,et al.Statin therapy accelerates reendothelialization:a novel effect involving mobilization and incorporation of bone marrow-derived endothelial progenitor cells.Circulation.2002;105(25):3017-3024.
[9]Yamahara K,Itoh H.Potential use of endothelial progenitor cells for regeneration of the vasculature.Ther Adv Cardiovasc Dis.2009;3(1):17-27.
[10]Kawamoto A,Asahara T.Role of progenitor endothelial cells in cardiovascular disease and upcoming therapies.Catheter Cardiovasc Interv.2007;70(4):477-484.
[11]Kalka C,Masuda H,Takahashi T,et al.Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization.Proc Natl Acad Sci U S A.2000;97(7):3422-3427.
[12]Tepper OM,Galiano RD,Capla JM,et al.Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation,adhesion,and incorporation into vascular structures.Circulation.2002;106(22):2781-2786.
[13]刘艳玲,李大勇,李雪晶.桃红四物汤调节动脉硬化闭塞症模型大鼠血液流动性的研究[J].中华中医药学刊,2014,32(4):761-763.
[14]张海峰,戴华卫,周瑞耀,等.酸性成纤维细胞生长因子对人脐带血内皮祖细胞凋亡的影响[J].中国动脉硬化杂志,2013,21(2):154-158.
[15]Peichev M,Naiyer AJ,Pereira D,et al.Expression of VEGFR-2 and AC133 by circulating human CD34(+)cells identifies a population of functional endothelial precursors.Blood.2000;95(3):952-958.
[16]王小斌,黄峻,朱莉,等.白藜三醇对人外周血内皮祖细胞功能的影响[J].江苏医学,2009,35(8):901-904.
[17]齐振熙,康靖东.桃红四物汤干预激素性股骨头缺血坏死组织局部微血管密度的变化[J].中国组织工程研究与临床康复,2009,13(11):2040-2043.
[18]龙翔,李娟,孙绍裘.桃红四物汤对兔皮片移植模型血管新生及MMP-2的影响[J].中医药导报,2016,22(22):14-17.
[19]池晓霞.桃红四物汤促进内皮细胞增殖及其机制初探[J].海峡药学,2010,22(1):39-41.
[20]刘竹青,尹登科,韩岚,等.桃红四物汤含药血清对过氧化氢损伤的人脐静脉内皮细胞的保护作用[J].中国中药杂,2013,38(3):402-406.
[21]Asahara T,Murohara T,Sullivan A,et al.Isolation of putative progenitor endothelial cells for angiogenesis.Science.1997;275(5302):964-967.
[22]Balbarini A,Barsotti MC,Di Stefano R,et al.Circulating endothelial progenitor cells characterization,function and relationship with cardiovascular risk factors.Curr Pharm Des.2007;13(16):1699-1713.
[23]Reyes M,Dudek A,Jahagirdar B,et al.Origin of endothelial progenitors in human postnatal bone marrow.J Clin Invest.2002;109(3):337-346.
[24]Werner N,Nickenig G.Influence of cardiovascular risk factors on endothelial progenitor cells:limitations for therapy.Arterioscler Thromb Vasc Biol.2006;26(2):257-266.
[25]Murayama T,Tepper OM,Silver M,et al.Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo.Exp Hematol.2002;30(8):967-972.
[26]Suzuki T,Nishida M,Futami S,et al.Neoendothelialization after peripheral blood stem cell transplantation in humans:a case report of a Tokaimura nuclear accident victim.Cardiovasc Res.2003;58(2):487-492.
[27]Vasa M,Fichtlscherer S,Aicher A,et al.Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease.Circ Res.2001;89(1):E1-7.
[28]Eizawa T,Ikeda U,Murakami Y,et al.Decrease in circulating endothelial progenitor cells in patients with stable coronary artery disease.Heart.2004;90(6):685-686.
[29]Loomans CJ,de Koning EJ,Staal FJ,et al.Endothelial progenitor cell dysfunction:a novel concept in the pathogenesis of vascular complications of type 1 diabetes.Diabetes.2004;53(1):195-199.
[30]Quyyumi AA.Circulating endothelial progenitor cells as novel biological determinants of vascular function and risk.Can JCardiol.2004;20 Suppl B:44B-48B.