| Microvascular injury is a common pathological change of acute myocardial infarction no-reflow,arrhythmia and chronic heart failure after myocardial infarction.The vessel-collateral theory proposes a concept:Ying and Wei rendezvous and transformation by collateral,it indicaes that its dysfunction lesion is the complex pathology of multidimensional spatiotemporal evolution of endothelial cells as the core and initiating factor,neurohumoral regulation and blood component participation,And the treatment strategy of "minute collateral-microvessel" protection was put forward,and the treatment strategy was adopted to the compound formation.For example,Tongxinluo capsule,Shensongyangxin capsule,Qiliqiangxin capsule,and their clinical effect is remarkable.The common protection to endothelial cell of the three compounds is the common mechanism in relevant diseasestreatment.Objective:The theoretical part of this study aims to explore the guidance of " Ying and Wei rendezvous and transformation by collateral" concept in relevant diseases treament,and put forward the "minute collateral-micro vessel" protection is the common intervention mechanism of the disease treament,focusing on the endothelium cell as the core of Tongluo-compounds research,To investigate whether Tongluo-compounds play a common role in endothelial cell protection and possible mechanism,and to further investigate if ginseng is the common material basis for Tongxinluo capsule,Shensongyangxin capsule or Qiliqiangxin capsule's protection effect to endothelial cell,and also the mechanism of barrier function protection in representative Tongluo-compounds to be further study.Through theoretical and experimental research to provide theoretical and experimental data support for the development of the vessel-collateral theory's guidance in relavent cardiovascular disease treatment.Methods:(1)Theoretical research:This research reviews the literature about Ying Wei and vessel-collateral,and puts forward "Ying and Wei rendezvous and transformation by collateral" concept,and which is used in the treatment of acute myocardial infarction no-reflow,arrhythmia and chronic Heart failure after myocardial infarction.Based on the concept "minute collateral-micro vessel" protection is proposed as a common intervention strategy of the above diseases.By analysing experimental research progress of Tongluo-compounds,we propose that the protection to endothelial cell is the core mechanism of Tongluo-compounds' common protection in "minute collateral-micro vessel".(2)Experimental research:Human umbilical vein endothelial cells(HUVEC)were cultured in vitro and the model was induced by Ox-LDL.?The survival status and survival activity were detected by MTS and live cell imaging.? PCR was used to detect the expression of eNOS mRNA and ET-1 mRNA.?The expression of total superoxide dismutase(SOD)in the cells was detected by WTS-8.? The expression of ROS was detected by flow cytometry.? The levels of IL-6 and sICAM were detected by enzyme-linked immunosorbent assay(ELISA).?The apoptosis rate was detected by flow cytometry.?The expression levels of Cyt C,Bax,Bcl-2,Fas,FasL,Caspase3 and Caspase8 were detected by Western blot.?Transwell chamber was used to detect the permeability of HUVEC cells monolayer.? The expression levels of ZO-1,Occludin,Claudinl,ERK1/2 and p-ERK1/2 were detected by Western blot.? The ?expression of tight junction protein was observed by laser confocal microscopy.Results:1.Theoretical research:? The research puts forward "Ying-Wei rendezvous and transformation by collateral" and provide a new strategy for the prevention and treatment of vascular disease.? Tongluo comounds have a common protection effect to "minute collateral-micro vessel",the protection to endothelial cells may be the common underlying mechanism for the effect.?The study of the endothelial cells within "Ying-Wei rendezvous and transformation by collateral" theory could be an important breakthrough point in the study the "minute collateral-micro vessel" protection of Tongluo-compounds.2.Experimental research:?Tongxinluo(TXL),Shensongyangxin(SS),Qiliqiangxin(QLQX),Ginsenosides(Zongzaogan,ZZ G),Ginsenoside Rbl and Leech(Shuizhi,SZ)have no drug toxicity within a certain range of drug concentration.TXL,SS can increase cell proliferation.? Tongluo compounds could significantly improve cell viability in Ox-LDL-induced HUVEC cells(P<0.05).?TXL,Rb1,SS and SZ could increase the expression of eNOS mRNA(P<0.05,P<0.01).QLQX and ZZG have no effect.TXL,SS,QLQX,ZZG,Rbl and SZ could decrease the expression of ET-1 mRNA(P<0.05,P<0.01).?TXL,QLQX,ZZG and Rb1 could decrease the level of IL-6(P<0.05,P<0.01),SS and SZ have no effect;TXL,QLQX,ZZG,Rbl,SZ could significantly reduce the level of sICAM(P<0.01),SS have no effect.?TXL,QLQX,SZ could increase the level of SOD(P<0.05),SS,ZZG,Rb1 have no effect;TXL,SS,QLQX and Rbl could decrease the level of ROS(P<0.05,P<0.01),and SZ have no effect.?TXL,SS,ZZG,Rbl and SZ could decrease the apoptosis of endothelial cells induced by Ox-LDL(P<0.05,P<0.01)and TXL effect is superior than SZ(P<0.05).?TXL,ZZG,Rbl could increase the ratio of Bcl-2/Bax(P<0.05,P<0.01),QLQX and SZ have no effect on Bcl-2/Bax ratio;TXL,ZZG and Rbl could decrease the expression of Cyt C(P<0.05).QLQX,SS and SZ have no effect.?TXL,SS,ZZG and Rbl could decrease the expression of Fas(P<0.01);TXL,SS,ZZG,Rbl and SZ could decrease the expression of FasL(P<0.05,P<0.01);TXL,ZZG and Rbl could decrease the expression of Caspase8(P<0.05,P<0.01);TXL,SS,ZZG,Rb1 and SZ could decrease the expression of Caspase3(P<0.05,P<0.01),and QLQX have no effect on any apoptosis-related proteins expression.?TXL,QLQX,SZ,ZZG could decrease the high permeability of Ox-LDL-induced monolayer endothelial cells(P<0.05,P<0.01).?TXL could increase the expression of tight junction protein ZO-1 and Occludin(P<0.05),but have no effect on C.laudinl expression;SS could decrease the high expression of Claudinl(P<0.01),but have no effect on the expression of ZO-1 and Occludin;QLQX could increase the expression of Occludin(P<0.05)and decrease the expression of Claudinl(P<0.01),but have no effect on the expression of ZO-1;ZZG could increase the expression of ZO-1 and Occludin(P<0.05,P<0.01),but no effect on Claudinl expression;Rbl could increase the expression of Occludin(P<0.01),but has no significant effect on the expression of ZO-1 and Claudinl;SZ could increase the expression of ZO-1(P<0.01),but no significant effect on Occludin and Claudinl expression.11TXL could decrease the high permeability of monolayer endothelial cells induced by Ox-LDL in a time-dependent manner(P<0.05,P<0.01).12TXL could increase the expression of ZO-1 and Occludin and improve the linear distribution of protein.13The phosphorylation of ERK1/2 was inhibited by PD98059 and TXL(P<0.01).14 PD98059 could significantly promote the expression of ZO-1(5?M),Occludin(5?M,10?M)protein at low dose(P<0.05),PD98059 could reduce the expression of Claudinl in a dose-dependent manner(P<0.05,P<0.01).15 TXL could significantly increase the expression of ZO-1 and Occludin in endothelial cells injured by Ox-LDL(P<0.01).PD98059 could partially inhibite the expression of ZO-1 in TXLpretreatment(P<0.05).The effect of TXL on Occludin was significantly inhibited by PD98059(P<0.01).16TXL could significantly reduce the high permeability of monolayer endothelial cells in Ox-LDL-induced cells,and PD98059 could reduce the effect of TXL on the permeability of monolayer endothelial cells(P<0.05,P<0.01).Conclusion:1.Based on the theory of "Ying-Wei rendezvous and transformation by collateral","Minute collateral-microvessel" protection is the common mechanism of Tongluo-compounds effect on acute myocardial infarction no-reflow,arrhythmia and chronic heart failure which appear after myocardial infarction.It enrichs the scientific connotation of vessel-collateral theory and makes a new guidance to cardiovascular disease prevention and treatment.2.Which probably an important reason for the protective effect of "minute collateral-micro vessel" is that Tongluo-compounds effect on endothelial cell protection in the prevention and treatment of cardiac "minute collateral-microvascular" disease.Which reflecting the different characteristics of endothelial protection due to different herb ingredients.Tongxinluo capsule has a wide range of endothelial cell protection,not only improve the endothelial cell inflammation oxidative damage,inhibition of endothelial cell apoptosis,and also can protect the endothelial cell structure,improve the damaged monolayer endothelial cell barrier fuction;Shensongyangxin capsule has a significant role in mproving endothelial cell oxidative damage and inhibition of apoptosis;Qiliqiangxin capsule can alleviate the oxidative damage of endothelial cells and protect the endothelial cell structure and improve the barrier function of monolayer endothelial cells.It can improve the endothelial cell damage and inhibit the apoptosis of endothelial cells.The protective effect of different Tongluo-compounds on endothelial cell provides data support to microvascular protection effect of Tongluo-compounds.3.Ginseng,which exist in Tongxinluo capsule,Shensongyangxin capsule and Qiliqiangxin capsul,has a protective effect on endothelial cell.Due to the different composition of the three compounds,the different effects of endothelial protective effect appeared,which suggesting that the composition of the compound may play.an importarnt role in the protection of endothelial cells,such as leeches also have a protectioin effect on endothelial cell,but its role in endothelial cell protection is not same to ginseng and Tongxinluo capsule.4.Tongxinluo capsule has a widely protective on endothelial cell,not only to improve the secretion of endothelial cells,inhibition of endothelial cell apoptosis,also can protect the endothelial cell structure,improve the endothelial cell barrier fimction.The study which could provide data support for the research on vascular protection effect of Tongxinluo capsule. |
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