| Background:The loss of the myocardial cell loss and ventricular remodeling was the pathological basis caused by heart failure and clinical death after acute myocardial infarction (AMI). The previous amount of literatures and our research mainly focused attention on the role of residual myocardial cell hypertrophy in myocardial remodeling after myocardial infarction, while the recent studies have put more mind on the role of necrotic interstitial fibrosis in myocardial remodeling after myocardial infarction and the excessive proliferation of muscle fibroblast was the central link of interstitial fibrosis. In addition, in the process of kidney, liver as well as lung fibro found that the excessive proliferation of myofibroblast not only was caused by the abnormal activation of fibroblast cell in site, but also got through epithelial -mesenchymal transition(EMT),while the research of the development of embryo heart have showed that the alternative form of the EMT was endothelial-mesenchymal transition(EndMT) and the mechanisms of the two types of fibrosis were contributed to the interstitial fibrosis after myocardial infarction, of which EndMT was mediated by the TGF-beta 1-Smad, while HGF could inhibit the EndMT by blocking the TGF?1 and the transformation of fibroblast into myofibroblast. Based on the findings above, we speculated that EndMT might also play a significant role in myocardial interstitial fibrosis after myocardial infarction. Provided that, we hypothesize, the one could be found to substitute for necrosis myocardial cells after myocardial infarction and secrete or carry HGF to inhibit EndMT as well as the way that fibroblast could transform myofibroblast as to inhibit myocardial interstitial fibrosis after myocardial infarction, which could provide the new research and treatment the improvement of myocardial remodeling and the prevention of the development of heart failure after myocardial infarction.Objective:To research the effect of adipose derived stem cells ADSCs) and HGF in the anoxia and ischemic model on the cardiac microvascular endothelial cells EndMT in rats is to provide the basis for clinical application of ADSCs combined with HGF in the cardiovascular fieldMethods:?. The Isolation, Culture as well as Identification of ADSCs To isolate and culture mesenchymal stem cells from the rats'fat, after subculture and amplification, was to monitor the cell activity and the cell phenotype to the proliferation induce of osteogenic adipogenesis.?. The Culture and Identification of Microvascular Endothelial Cells To isolate and culture microvascular endothelial cells from cardiac tissues, after subculture and amplification, were to monitor the call activity and identify Immunofluorescence staining.?. Establishment of the anoxia and ischemic model of microvascular endothelial cells in rat, as well as the effect of ADSCs and HGF on microvascular endothelial cells EndMT in this modelSubculturing 2 generations of micro vascular endothelial cells were seeded in 35 mm culture dish, after integrated into the complete cell monolayer, to flush two times with sterile buffer Hank, then add buffer containing calcium as well as low sugar to gently drip the aseptic paraffin oil into the Eaele buffer, of which was covered completely in the Eaele liquid, place it under 37?, and culture the small culture dish placed in incubator. In addition, the experiment was randomized 2 groups:shame group and ADSCs-HGF group to gather the supernatant after interference 4 days as to determine respectively the expression of the supernatant a-SMA and viment applied ELISA and Western blot.?.To treat rat with acute myocardial infarction (AMI) by transplant in the model of ADSCs-HGF, the SD rat was randomized two groups:shame group that injected the DMEM on the edge of the infarct area of epicardial with sterile micro syringe, each 50 ml, after AMI, and DSCs-HGF group injected ADSCs as well as HGF with the same approach, each 50 ml (cell number 2.5 x 10 ml/50) and obtained the cardiac tissues to test double immunofluorescence staining in the postoperative period for 1 month..Result:The successful isolation and culture of ADSCS showed that CD29. CD90 CD31, CD34 as well as CD45 were positive by identification and successfully differentiated the bone, fat, myocardial cell by induction, of which its success in rat infants identified endothelial cells by PAP. In addition, the successful establishment of microvascular endothelial cells in ischemia hypoxia model observed the loss of a-SMA of adipose derived stem cells and HGF and the expression of vimentin (P< 0.05) in this model.Conclusion:1. this experiment successfully isolated adipose derived stem cells (ADSCs) and mice cardiac microvascular endothelial cells.2. ADSCs combined with HGF could inhibit the differentiation of endothelial cells into fibroblasts... |
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