Effects And Mechanism Of Trichosanthes On The Regulation Of The Motivation Of Endothelial Progenitor Cells In Coronary Heart Disease

Objective: This project intends to study the pharmacodynamical effect ofTrichosanthes pericarpium in Coronary heart disease using animal model as well as murinebone marrow-derived EPCs in vitro. Study the role of Trichosanthes pericarpium onendothelial progenitor cell mobilization, and reveal the effect and mechanism ofTrichosanthes pericarpium on coronary heart disease.Methods:(1)Wistar male rats were randomly divided into the control group, modelgroup, the danshenpian group (positive control group), the trichosanthes group.To makeacute myocardial ischemia animal model，1hour after the last administration, the leftanterior descending coronary branch were ligated in male Wistar rats that have receivedintragastrical administration for4days.6h after of operation, myocardial tissue wasstained with triphenyltetrazolium chloride(TTC) to calculate the infarction rate. Toevaluate the myocardial injury, serum levels of creatine-kinase(CK), creatine-kinase-MB(CKMB), lactate dehydrogenase(LDH),α-Hydroxybutyrate Dehydrogenase(α-HBDH) andaspartate-amino transferase (AST) were detected in these different groups of rats.(2) Another batch of Wistar male rats were randomly divided into the control group,model group and the trichosanthes group, Flow cytometric analysis was performed toquantify the circulating EPCs.(3) Mononuclear cells (MNCs) in murine bone marrow were isolated, and then thecells were cultured in M199medium supplemented with VEGF and bFGF. Establishischemic and hypoxic model of cultured bone marrow-derived endothelial progenitor cellsof rats in vitro; In this model, cultured EPCs was subjected to insufficient blood and oxygen environment;then add Gualoupi injection in culture medium, MTT assay was usedto evaluate the effects of trichosanthes on the proliferative capacity of EPCs．Theconcentration of VEGF in culture medium was measured by enzyme-linkedimmunosorbent assay (ELISA) to investigate the effects of Trichosanthis on mobilizationof EPCs.Results:(1) Compared with that in the model group rats, the myocardial infarctionrate of rats in the Trichosanthes group decreased evidently (P<0.05), the serum activity ofCK, CK-MB, LDH, AST and α-HBDH were also obviously low (P<0.05or P<0.01).(2) The number of circulating endothelial progenitor cells (CEPC) of the model ratsincreased in the first3days, and was reduced slightly on the fourth day. Myocardialischemia is one important factor that stimulate the mobilization of EPCs; The number ofEPC in trichosanthes group upgraded from the first day and is higher than the model groupon the fourth day．The trichosanthes can promote the mobilization of EPCs.(3) Mononuelear cells(MNCs) isolated from the bone marrow turned tospindle-shaped,endothelial cell like cells after7days culture. EPCs were characterized asadherent cells with double positive for DiLDL-uptake and lectin binding(by directfluorescent staining)under a laser scanning confocal microscope(LSCM). According to theresults of MTT assay, Trichosanthes can enhance the proliferative capacity of EPCs, Theproliferative capacity of EPCs improved (P<0.01)，with a peak at concentration of20mg/ml(P<0.01).Conclusion：The protective effect of Trichosanthes pericarpium on the rats acutemyocardial ischemic is significant. The Trichosanthes pericarpium can promote themobilization of EPCs. Trichosanthes can enhance the proliferative capacity of EPCs, andcan promote the release of VEGF in cell supernatant, the protective effect may relate to thepromotion of the VEGF release.