The Research On Vascular Endothelial Cell (VEC) Injury Model Of Blood Stasis Syndrome (BSS) Associated With Diabetis

ObjectiveTo establish the vascular endothelial cell(VEC) injury model of syndrome(BSS) associated with diabetes(DB),to observe the model cells' change of cytoactive,shape,ultrastructure and function,and to provide reference in the methodology for the establishment of traditional Chinese medicine(TCM) cell model based on integration of disease and syndrome.Partâ… .Research on the injury model by the serum of DB patients with BSS1.Effects of ECV-304 incubated by the serum of patients with BSS associated with diabetes[Objective]To observe the effects of ECV-304 incubated by the serum of patients with BSS associated with diabetes.[Methods]ECV-304 were obtained by cultured conventionally.It was divided into 4 groups,ECV-304 cultured with DMEM(Group N),with the serum of healthy person(Group H),with the serum of patients with BSS associated with diabetes(Group MD) and the serum of diabetes patients with no-BSS(Group MND).The cytoactive of ECV-304 were assayed by MTT chromatometry,and morpHological changes were identified by light microscope and electron microscopy.All groups were measured.[Results]â‘ There was a statistically significant difference(P＜0.001)regarding the OD in the MD Group and H Group.â‘¡By the inverse phase contrast microscope,we discovered a significant difference between the Group MD and Group MND.[Conclusion]The serum of patients with BSS associated with diabetes could injury ECV-304 of the cytoactive and shape.2.Preliminary research on the mechanisms of injury ECV-304 by the serum of patients with BSS associated with diabetes on endothelial dysfunction[Objective]To explore the mechanisms of ECV-304 incubated by the serum of patients with BSS associated with diabetes.[Methods]ECV-304 was divided into 4 groups the same as the 1^st experiment.The nitric oxide(NO) excreted by model cells was assayed by nitric acid deoxidizing enzyme method,and the endothelin(ET) was assayed by non-balance method.The injury symbols of VEC:vWF,TM and EPCR excreted by model cells were assayed by enzyme-linked immunosorbent assay (ELISA).All groups were measured.[Results]â‘ There was a significant difference(P＜0.001) regarding the ET excreted by ECV-304 between the Group MD and Group H,Group N,Group MND.â‘¡There was a significant difference(P＜0.001) regarding the NO excreted by ECV-304 between the Group MD and Group N,Group H.â‘¢There was a significant difference(P＜0.001) regarding the EPCR excreted by ECV-304 between the Group MD and Group N,Group H,Group MND.â‘£There was a significant difference(P＜0.001) regarding the vWF excreted by ECV-304 between the Group MD and Group N.⑤And the same condition of TM happened between the Group MDand the Group N significantly(P＜0.001).[Conclusion]The serum of patients with BSS associated with diabetes could injury ECV-304 and caused endothelial dysfunction.It was possible that BSS influnenced the expression of molecules in vascular endothelial cells.3.Preliminary research on the mechanisms of injury ECV-304 by the serum of patients with BSS associated with diabetes on intracellular free calcium([Ca²⁺]i) and cytoskeleton[Objective]To explore the mechanisms of ECV-304 incubated by the serum of patients with BSS associated with diabetes.[Methods]ECV-304 was divided into 4 groups the same as the 1^st experiment.Model cells' intracellular free calcium([Ca²⁺]i) and cytoskeleton were assayed by laser scanning confocal microscope(LSCM).All groups were measured.[Results]â‘ There was a significant difference(P＜0.001) regarding the intracellular calcium concentration([Ca²⁺]i) of ECV-304 between the Group MD and Group H,Group N,Group MND.â‘¡by the cytoskeleton,the Group MD was distinctly different with the Group MND.[Conclusion]The serum of patients with BSS associated with diabetes could injury ECV-304 and caused Model cells' intracellular free calcium([Ca²⁺]i) and cytoskeleton change.4.Protective effects of Xuefn Zhuyu decoction and Tanâ…¡A on the injury model by the serum of patients with BSS associated with diabetes on endothelial dysfunction [Objective]To observe the protective effectes of Xuefu Zhuyu decoction and Tanâ…¡A on the cellar injury model by the serum of patients with BSS associated with diabetes on endothelia dysfunction.[Methods]ECV-304 were obtained by cultured conventionally.It was divided into 4 groups,Group N and Group MD is the same as 1^st experiment,the group incubated with the serum of patients with BSS associated with diabetes and the mice serum of Xuefu Zhuyu decoction(Group MDB),the group with the serum of diabetes patients with BSS and Tanâ…¡A(Group MDT).The activeness of ECV-304 were assayed by MTT chromatometry,and morpHological changes were identified by light microscope and electron microscopy.The nitric oxide(NO) excreted by model cells was assayed by nitric acid deoxidizing enzyme method,and the endothelin(ET) was assayed by non-balance method.The injury symbols of VEC:vWF,TM and EPCR excreted by model cells were assayed by enzyme-linked immunosorbent assay(ELISA).Model cells' intracellular free calcium ([Ca²⁺]i) and cytoskeleton were assayed by laser scanning confocal microscope(LSCM).All groups were measured.[Results]â‘ By the inverse pHase contrast microscope,we discovered that the Group MDB and Group MDT is the same shape as the Group N.â‘¡There was a significant difference(P＜0.001)of the secretion of OD,NO,ET,EPCR,vWF,TM and intracellular calcium concentration between the Group MDB and the Group MD.â‘¢There was a significant difference(P＜0.001) of the secretion of OD,NO and ET between the Group MDT and the Group MD,but no differences in the intracellular calcium concentration and the injury symbols.â‘£by the cytoskeleton,the Group MDB and Group MDT were distinctly different with the Group MD.[Conclusion]The Xuefu Zhuyu decoction and Tanâ…¡A had the protective effectes on the cellar injury model by the serum of patients with BSS associated with diabetes on endothelia dysfunction and from the point of medicine,they could recover endothedial dysfunction induced by the cellar injury model the medicine.Partâ…¡.Differences between injury models by the serum of EH patients with BSS and by the serum of DB patients with BSS [Objective]To investigate the differences between injury models by the serum of EH patients with BSS and by the serum of DB patients with BSS.[Methods] ECV-304 were obtained by cultured conventionally.It was divided into 4 groups. Group N,Group H and Group MD is the same as 1^st experiment,the group incubated with the serum of patients with BSS associated with EH(Group MH).The activeness of ECV-304 were assayed by MTT chromatometry,and morpHological changes were identified by light microscope and electron microscopy.The nitric oxide(NO) excreted by model cells was assayed by nitric acid deoxidizing enzyme method,and the endothelin(ET) was assayed by non-balance method.The injury symbols of VEC: vWF,TM and EPCR excreted by model cells were assayed by enzyme-linked immunosorbent assay(ELISA).Model cells' intracellular free calcium([Ca²⁺]i) and cytoskeleton were assayed by laser scanning confocal microscope(LSCM).All groups were measured.[Results]There was a significant difference of the secretion of OD, NO,ET,EPCR,vWF,intracellular calcium concentration and cytoskeleton between the Group MD and the Group MH.[Conclusion]The results clearly show that the changes between the Group MD and the Group MH is partly related to the basis of BSS.Furthermore,by the example of BSS,it can partially explain the mechanism of "different disease and same syndrome" in Traditional Chinese Medicine.