How does autophagy activation affect the apoptosis, proliferation and cycle of endothelial progenitor cells in rats? / 中国组织工程研究

BACKGROUND:Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cels, but there is no report on the effect of autophagy, as wel as the interaction between autophagy and apoptosis. OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cels. METHODS:Density gradient centrifugation was used to obtain mononuclear cels from bone marrow, and the mononuclear cels were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cels colected were the endothelial progenitor cels. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cel cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cels in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-IIprotein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagywas particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cels was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cel apoptosis, change the cel cycle significantly, and can inhibit the proliferation of endothelial progenitor cels.

The effect of 3-MA on proliferation, apoptosis and cell cycle of rat endothelial progenitor cells / 中华普通外科杂志

Objective To investigate the effect of proliferation,apoptosis and cell cycle of 3-MA on rat endothelial progenitor cells. Methods Bone marrow-derived mononuclear cells were isolated from rat bone marrow by ficoll. There were five groups. The control group and four 3-MA concentration groups: 1. 25 mmol/L,2. 5 mmol/L,5 mmol/L, 10 mmol/L. MTT was used to measure the proliferation of endothelial progenitor cells. Flow cytometry ( FCM) was used to detect cell apoptosis and cell cycle. Results (1)5 mmol/L 3-MA promotes proliferation of endothelial progenitor cells, while 10 mmol/L 3-MA inhibits the proliferation of endothelial progenitor cells (P ＜ 0. 05). (2) 10 mmol/L 3-MA promotes apoptosis of endothelial progenitor cells, compared with the control, the difference was significant ( P ＜ 0. 05 ). (3) 3-MA at the concentration of 5 mmol/L reduces cells at G0/G1 phase and increases S and G2/M phase cells; 10 mmol/L 3-MA induces endothelial progenitor cells blockade at S phase, G2/M phase cells decreased, compared with the control, the difference was significant (P ＜ 0. 05). Conclusions 5 mmol/L 3-MA promotes the proliferation of endothelial progenitor cells. 10 mmol/L 3-MA inhibits the proliferation and promotes apoptosis of endothelial progenitor cells.

Interventional therapy and its efficacy for lower extremity arteriosclerosis occlusive disease of 36 cases / 中国综合临床

Objective To explore the clinical application of pereutaneous transluminal angioplasty(PTA) and endovascular bracket to treat lower extremity arteriosclorotic occlusion.Methods The clinical data of 36 patients(41 affected limbs)with lower extremity atherosclerotic occlusion who were treated with PTA and bracket implantation from Jan 2008 to Dee 2008 were summarized.Results The initial successful rate of PTA wag 95.1% (39/41).The clinical symptoms were considerably improved in 37 affected limbs,representing of pain disappearing,skin temperature increasing and the healing of refractory ulcer.The index of ankle to brachial significantly increased from 0.54±0.11 to 0.79±0.15(before v.s.after therapy).However,no improvement was observed in 3 affected limbs,and one affected limb Wag re-operated by the amputation.In the following 3 to 15 months.three superficial femorsI arteries were re-obstructed at the 5th,6th,12th month,respectively.One arteria tibialis pesterior was re-obstructed at the 8th month.The cumulative cure rate was 89.7%(35/39).Conclusions PTA is effective in treating atherosclerotic occlusive diseases.The endovascular bracket can increase the cumulative cure rate.PTA and endovascular bracket are safe and effective in treating lower extremity arteriosclerotie occlusion.