益气活血方对气虚血瘀证大鼠血管内皮功能的影响
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摘要
目的:探讨气虚血瘀证大鼠血管内皮细胞功能变化及益气活血方(YQHXF)对其的影响。方法:实验分为空白组,气虚血瘀组,YQHXF高、低剂量组(5. 54,2. 77 g·kg-1)。采用水环境小平台站立长期无规律、不完全的睡眠剥夺法,每天剥夺16 h,持续6周,制造大鼠神劳加体劳的双重消耗建立大鼠气虚血瘀证候模型,自第5周开始灌胃给药,连续给药2周至实验结束。酶联免疫吸附测定(ELISA)检测大鼠血浆中内皮素-1(ET-1),血管性血友病因子(v WF),血管细胞黏附分子-1(VCAM-1),细胞间黏附分子(ICAM-1),P选择素,白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α)含量,硝酸还原酶法检测一氧化氮(NO)水平。结果:与空白组比较,气虚血瘀组大鼠呈现毛发粗糙晦暗,体质量、脉搏幅度显著降低(P <0. 01),舌面R,G,B值显著降低(P <0. 01),全血黏度高、中切值,血浆黏度均显著增高(P <0. 01),ET-1,v WF,VCAM-1,ICAM-1,P选择素,IL-6,TNF-α等血管内皮功能相关指标明显升高(P <0. 05,P <0. 01);与气虚血瘀组比较,YQHXF高、低剂量可显著升高气虚血瘀证大鼠脉搏幅度及舌面R,G,B值(P <0. 01),明显降低全血黏度高、中、低切值及血浆黏度(P <0. 05,P <0. 01),同时明显降低大鼠ET-1,IL-6,VCAM-1,P选择素水平(P <0. 05,P <0. 01)。结论:睡眠剥夺法可诱导大鼠气虚血瘀证形成,并可导致血管内皮功能失调。YQHXF具有保护血管内皮的功能,通过调节血管内皮活性物质分泌、减少细胞黏附、抑制炎症等途径改善气虚血瘀证大鼠气虚及血瘀症状。
Objective: To explore the changes and benefits of vascular endothelial cell function in rats with qi deficiency and blood stasis syndrome,and the effect of Yiqi Huoxue recipe( YQHXF) of such changes.Method: Rats were randomly divided into blank control group,Qi deficiency and blood stasis group,and YQHXF high and low dose groups( 5. 54,2. 77 g·kg-1). A small platform of water environment was used to make the rats stand for a long-term with irregular and incomplete sleep deprivation,16 h per day for six weeks,so that both mentality and labor of rats were consumed to establish qi deficiency and blood stasis rat models. From the fifth week,intragastric administration was given for 2 weeks,until end of the experiment. Then levels of endothelin-1( ET-1),von willebrand factor( vWF),vascular cell adhesion molecule-1( VCAM-1),intercellular adhesion molecule( ICAM-1),P-selectin,interleukin-6( IL-6),and tumor necrosis factor-α( TNF-α) in rat plasma were detected by enzyme-linked immunosorbent assay( ELISA) and nitric oxide( NO) was detected by nitrate reductase assay. Result: Compared with blank control group,rats in Qi deficiency and blood stasis group showed rough and dark hair,with significantly decreased body weight and pulse amplitude( P < 0. 01); R,G,B values of the tongue surface were decreased significantly( P < 0. 01); high and moderate shear values of whole blood viscosity,as well as plasma viscosity were significantly increased( P < 0. 01),and the indexes of vascular endothelial function such as ET-1,vWF,VCAM-1,ICAM-1,P-selectin,IL-6 and TNF-α were abnormally increased after sleep deprivation( P < 0. 05,P < 0. 01). High and low dose YQHXF significantly increased the pulse amplitude and tongue R,G,B values( P < 0. 01),reduced the high,moderate and low shear values of whole blood viscosity,as well as plasma viscosity( P < 0. 05,P < 0. 01); at the same time,the levels of ET-1,IL-6,VCAM-1 and P-selectin in rats were decreased( P < 0. 05,P < 0. 01). Conclusion: Sleep deprivation can induce the formation of Qi deficiency and blood stasis syndrome in rats,and lead to vascular endothelial dysfunction. YQHXF has the function of protecting the vascular endothelium. It can improve the Qi deficiency and blood stasis symptoms in rats with Qi deficiency and blood stasis syndrome by regulating the secretion of vascular endothelial active substances,reducing cell adhesion and inhibiting inflammation.
Objective: To explore the changes and benefits of vascular endothelial cell function in rats with qi deficiency and blood stasis syndrome,and the effect of Yiqi Huoxue recipe( YQHXF) of such changes.Method: Rats were randomly divided into blank control group,Qi deficiency and blood stasis group,and YQHXF high and low dose groups( 5. 54,2. 77 g·kg-1). A small platform of water environment was used to make the rats stand for a long-term with irregular and incomplete sleep deprivation,16 h per day for six weeks,so that both mentality and labor of rats were consumed to establish qi deficiency and blood stasis rat models. From the fifth week,intragastric administration was given for 2 weeks,until end of the experiment. Then levels of endothelin-1( ET-1),von willebrand factor( vWF),vascular cell adhesion molecule-1( VCAM-1),intercellular adhesion molecule( ICAM-1),P-selectin,interleukin-6( IL-6),and tumor necrosis factor-α( TNF-α) in rat plasma were detected by enzyme-linked immunosorbent assay( ELISA) and nitric oxide( NO) was detected by nitrate reductase assay. Result: Compared with blank control group,rats in Qi deficiency and blood stasis group showed rough and dark hair,with significantly decreased body weight and pulse amplitude( P < 0. 01); R,G,B values of the tongue surface were decreased significantly( P < 0. 01); high and moderate shear values of whole blood viscosity,as well as plasma viscosity were significantly increased( P < 0. 01),and the indexes of vascular endothelial function such as ET-1,vWF,VCAM-1,ICAM-1,P-selectin,IL-6 and TNF-α were abnormally increased after sleep deprivation( P < 0. 05,P < 0. 01). High and low dose YQHXF significantly increased the pulse amplitude and tongue R,G,B values( P < 0. 01),reduced the high,moderate and low shear values of whole blood viscosity,as well as plasma viscosity( P < 0. 05,P < 0. 01); at the same time,the levels of ET-1,IL-6,VCAM-1 and P-selectin in rats were decreased( P < 0. 05,P < 0. 01). Conclusion: Sleep deprivation can induce the formation of Qi deficiency and blood stasis syndrome in rats,and lead to vascular endothelial dysfunction. YQHXF has the function of protecting the vascular endothelium. It can improve the Qi deficiency and blood stasis symptoms in rats with Qi deficiency and blood stasis syndrome by regulating the secretion of vascular endothelial active substances,reducing cell adhesion and inhibiting inflammation.
引文
[1]王嘉麟,郭蓉娟,张允岭,等.脑梗死恢复期气虚血瘀证患者中医临床路径评价与思考[J].环球中医药,2012,5(12):908-910.
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[10]李迎春.中药补阳还五汤治疗脑梗塞气虚血瘀证的效果研究[J].智慧健康,2018,4(12):76-77.
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[12]史雅红,任建勋,刘建勋,等.睡眠剥夺诱导的大鼠血瘀证对血小板活化影响的研究[J].中国中药杂志,2018,43(16):3341-3345.
[13]郝婷婷,郭浩,刘建勋,等. 2种分析方法对睡眠剥夺大鼠血瘀证评价的初步研究[J].中国中药杂志,2018,43(9):1880-1885.
[14]刘建勋.病症结合动物模型拟临床研究思路与方法[M].北京:人民卫生出版社,2014:70-76.
[15]陈利国,胡小勤.论病证结合血瘀证血管内皮细胞损伤模型的建立[J].中国中西医结合杂志,2007,27(3):267-269.
[16] Maier J A. Endothelial cells and magnesium:implications in atherosclerosis[J]. Clinl Sci,2012,122(9):397-407.
[17] Akiko H,Takahisa T,Hiroko M,et al. Advanced glycation end products increase endothelial permeability through the RAGE/Rho signaling pathway[J]. Febs Lett,2010,584(1):61-66.
[18]莫莉.冠心病气虚血瘀证的临床特征及益气活血法干预的实验研究[D].长沙:湖南中医药大学,2008.
[19]杨宝元,李继安,董印宏,等.冠心病气虚血瘀证血管内皮细胞功能失调相关性研究[J].江苏中医药,2011,43(8):28-30.
[20]荣晓凤,李荣亨,吕霞.气虚血瘀证大鼠NO、ET、TXB2、6-keto-PGF1α的变化及意义[J].中国中医基础医学杂志,2009,15(12):914-916.
[21]李艳,郑桂玲.气虚血瘀证与冠心病血浆一氧化氮、内皮素和血管紧张素Ⅱ含量的相关性研究[J].成都中医药大学学报,2012,35(4):11-12.
[22] Kanaji N,Sato T, Nelson A, et al. Inflammatory cytokines regulate endothelial cell survival and tissue repair functions via NF-κB signaling[J]. J Inflamm Res,2011,4(1):127-138.
[23]林桂永,王子健.冠心病血瘀证气血辨证与内皮细胞损伤的关系[J].中西医结合心脑血管病杂志,2005,3(10):856-858.
[24]陈海燕.麝香保心丸对气虚血瘀型冠心病患者PCI术后心肌损伤保护作用的临床研究[D].福州:福建中医药大学,2012.
[25] Jansen M P B,Florquin S,Roelofs J J T H. The role of platelets in acute kidney injury[J]. Nat Rev Nephrol,2018,14(7):457-471.
[26]李艳,刘卫星.血浆s ICAM-1、s VCAM-1含量与冠心病气虚血瘀证的相关性研究[J].山东中医药大学学报,2012,36(6):487-488.
[2]朱明军,李彬,王永霞.充血性心力衰竭中医病因病机分析[J].世界中西医结合杂志,2009,4(1):1-2.
[3]赵爱梅,任钧国,刘建勋.益气活血方治疗冠心病气虚血瘀证作用机制研究进展[J].中国实验方剂学杂志,2017,23(7):215-220.
[4]于赫,孙力.气虚证的生物化学与分子生物学研究进展[J].中医药信息,2005,22(5):48-51.
[5] Schiro A,Wilkinson F,Weston R,et al. Endothelial microparticles as conveyors of information in atherosclerotic disease[J]. Atherosclerosis,2014,234(2):295-302.
[6] Favero G,Paganelli C,Buffoli B,et al. Endothelium and its alterations in cardiovascular diseases:life style intervention[J]. Biomed Res Int,2014, 2014(2):801896.
[7] Tarone G,Balligand J L,Bauersachs J,et al. Targeting myocardial remodelling to develop novel therapies for heart failure:a position paper from the working group on myocardial function of the european society of cardiology[J]. Eur J Heart Fail,2014,16(5):494-508.
[8] ZHANG H,WANG W R, LIN R, et al. Buyang Huanwu decoction ameliorates coronary heart disease with Qi deficiency and blood stasis syndrome by reducing CRP and CD40 in rats[J]. J Ethnopharmacol,2010,130(1):98-102.
[9]石立鹏,杜旭勤,赵凤林,等.针刺联合补阳还五汤治疗气虚血瘀证冠心病稳定型心绞痛疗效及对血脂水平的影响[J].现代中西医结合杂志,2017,26(35):3880-3882,3966.
[10]李迎春.中药补阳还五汤治疗脑梗塞气虚血瘀证的效果研究[J].智慧健康,2018,4(12):76-77.
[11]陈进成,刘建勋,林成仁,等.基于“劳则气耗”理论研究气虚证动物模型的建立方法[J].中国中药杂志,2018,43(11):2177-2183.
[12]史雅红,任建勋,刘建勋,等.睡眠剥夺诱导的大鼠血瘀证对血小板活化影响的研究[J].中国中药杂志,2018,43(16):3341-3345.
[13]郝婷婷,郭浩,刘建勋,等. 2种分析方法对睡眠剥夺大鼠血瘀证评价的初步研究[J].中国中药杂志,2018,43(9):1880-1885.
[14]刘建勋.病症结合动物模型拟临床研究思路与方法[M].北京:人民卫生出版社,2014:70-76.
[15]陈利国,胡小勤.论病证结合血瘀证血管内皮细胞损伤模型的建立[J].中国中西医结合杂志,2007,27(3):267-269.
[16] Maier J A. Endothelial cells and magnesium:implications in atherosclerosis[J]. Clinl Sci,2012,122(9):397-407.
[17] Akiko H,Takahisa T,Hiroko M,et al. Advanced glycation end products increase endothelial permeability through the RAGE/Rho signaling pathway[J]. Febs Lett,2010,584(1):61-66.
[18]莫莉.冠心病气虚血瘀证的临床特征及益气活血法干预的实验研究[D].长沙:湖南中医药大学,2008.
[19]杨宝元,李继安,董印宏,等.冠心病气虚血瘀证血管内皮细胞功能失调相关性研究[J].江苏中医药,2011,43(8):28-30.
[20]荣晓凤,李荣亨,吕霞.气虚血瘀证大鼠NO、ET、TXB2、6-keto-PGF1α的变化及意义[J].中国中医基础医学杂志,2009,15(12):914-916.
[21]李艳,郑桂玲.气虚血瘀证与冠心病血浆一氧化氮、内皮素和血管紧张素Ⅱ含量的相关性研究[J].成都中医药大学学报,2012,35(4):11-12.
[22] Kanaji N,Sato T, Nelson A, et al. Inflammatory cytokines regulate endothelial cell survival and tissue repair functions via NF-κB signaling[J]. J Inflamm Res,2011,4(1):127-138.
[23]林桂永,王子健.冠心病血瘀证气血辨证与内皮细胞损伤的关系[J].中西医结合心脑血管病杂志,2005,3(10):856-858.
[24]陈海燕.麝香保心丸对气虚血瘀型冠心病患者PCI术后心肌损伤保护作用的临床研究[D].福州:福建中医药大学,2012.
[25] Jansen M P B,Florquin S,Roelofs J J T H. The role of platelets in acute kidney injury[J]. Nat Rev Nephrol,2018,14(7):457-471.
[26]李艳,刘卫星.血浆s ICAM-1、s VCAM-1含量与冠心病气虚血瘀证的相关性研究[J].山东中医药大学学报,2012,36(6):487-488.
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