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[ ABSTRACT ] AIM: To detect the autophagic changes of human pulmonary microvascular endothelial cells ( HPMVECs) under ischemia/reperfusion ( I/R) microenvironment, and to clarify the effects of autophagy on the HPM-VECs survival and endothelial barrier integrity under I/R condition.METHODS:Rapamycin ( RAP) was applied to pro-mote autophagy of HPMVECs.These cells were then incubated under the condition of oxygen-glucose deprivation/oxygen-glucose restoration ( OGD) .After exposure to OGD, the changes of autophagy, cellular death and permeability of the cells were determined by transmission electron microscopy, flow cytometry and transwell assay, respectively.RESULTS:Com-pared with the control cells, OGD-challenged cells had a much higher level of autophagy.The apoptotic rate was much higher and endothelial permeability was more serious in OGD group than those in control group.Preconditioning with RAP effectively improved OGD induced autophagy, it did not affect the cell survival and endothelial permeability under normal living condition, but obviously decreased the cells apoptotic rate, and remarkably lowered OGD-induced high permeability of the cells.CONCLUSION:Autophagy protects HPMVECs against I/R-induced injury.Promotion of autophagy is help-ful for attenuating I/R-induced cell death and sustaining the endothelial barrier integrity.
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AIM: To detect the changes of autophagy in adipose cells under starvation , and to clarify the effects of autophagy on the cell survival and apoptosis under starvation .METHODS: Rapamycin ( RAP) was applied to promote autophagy of adipose cells .These cells were then incubated under oxygen-glucose deprivation ( OGD) condition. After exposure of the cells to OGD , the changes of autophagy and apoptosis were determined by Western blotting , transmis-sion electron microscopy and TUNEL assay .RESULTS:Compared with the control cells , OGD-challenged cells had much higher level of autophagy .The apoptotic rate in OGD group was much higher than that in control group , which was reflected by increased protein level of activated caspase-3 and percentages of TUNEL positive cells .Preconditioning with RAP effec-tively improved OGD-induced autophagy , but did not affect the cell survival and apoptosis under normal condition , and ob-viously decreased the apoptotic rate of the cells under OGD condition .CONCLUSION:Autophagy protects adipose cells against starvation-induced apoptosis .Promotion of autophagy is helpful for attenuating starvation-induced apoptosis of the cells under OGD condition .
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Objective To investigate the feasibility of transplantation of mesenchymal stem cell (aisc) into expanion skin. Methods MSCs were isolated from porker's bone marrow and cultured in vitro. Pigs were randomly divided into four groups: Group A was injected with MSCs on the local expansion; Group B was injected with MSCs from ear vein; Group C was planted expander only; Group D was the contronl group. Each side of pig's spinal column was implanted with three expander in groups A, B and C. The same volume of NS was injected at the fixed time, the marked area measured after 7, 14, 28 days of expansion, the difference of those area was compared between groups. The differentiation of BM-MSC was detected by immunofluorescence. Results Flow-cytometric analysis showed that these BMSCs expressed CD90 and CD29 highly but did not express CD34 or CD45. The expansion area (cm2) of groups A, B, C and D was 34.05±0. 92, 31.83±l. 07,30. 10±0.79, and 18. 27±0.25, respectively (P＜0. 01). Immunofluorescence showed that the positive expression rate of CD31, PCNA in groups A and B was higher than that in groups C and D, in which the expression was the highest in group A. Conclusions Allogeneic transplantation of BM-MSC can promote skin expansion and the effect of local transplantation group is most significant.