ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of cyclosporine A-nanoparticles emulsion (CsA-NP) on protecting apoptosis of swine adipose tissue-derived stem cells (ASC ) and related mechanisms.</p><p><b>METHODS</b>ASC were randomized to six groups: control group,single H2O2 group,CsA or CsA-NP 0.1 mg/ml+H2O2 group,CsA or CsA-NP 1.0 mg/ml+H2O2 group, CsA or CsA-NP 5.0 mg/ml+H2O2 group,CsA or CsA-NP 10.0 mg/ml+H2O2 group. ASC apoptosis was induced by hydrogen peroxide (H2O2100 µmol/L) in vitro. The morphology of apoptotic cells was observed and the number of apoptotic cells was measured. Apoptosis of ASC was detected by flow cytometry using an apoptosis kit. Cell activity was determined by CCK-8 assay. Caspase-3 activity was detected by applying a caspase-3 assay kit. Expression of cytochrome C was investigated by Western blot.</p><p><b>RESULTS</b>H2O2 induced ASC apoptosis was evidenced by morphological and biochemical changes,which could be significantly reduced by pre-treatment with CsA or CsA-NP at concentration of 0.1-10.0 mg/ml, and the best effect was observed at concentration of 5 mg/ml (apoptosis rate: CsA: 10.6% ± 2.8% vs. 25.2% ± 3.8%; CsA-NP: 6.2% ± 2.6% vs. 25.2% ± 3.6% in control group, all P < 0.01). The cell activity was significantly higher in CsA or CsA-NP pre-treated ASC at concentration of 0.1-10.0 mg/ml than in H2O2 group (P < 0.01). Pre-treatment with CsA or CsA-NP (0.1-10.0 mg/ml) significantly down -regulated caspase-3 activity. Furthermore, CsA or CsA-NP (5 mg/ml) completely inhibited the H2O2-induced release of cytochrome C.</p><p><b>CONCLUSIONS</b>These results suggest that CsA-NP and CsA could protect the oxidative stress-induced ASC apoptosis through decreasing the activation of caspase-3 and inhibiting the release of cytochrome C.</p>
Subject(s)
Animals , Adipose Tissue , Cell Biology , Apoptosis , Cells, Cultured , Cyclosporine , Pharmacology , Nanoparticles , Stem Cells , Pathology , SwineABSTRACT
<p><b>OBJECTIVE</b>To evaluate the efficacy of cyclosporine A-nanoparticles emulsion (CsA-NP) combined with adipose tissue-derived stem cells (ASCs)transplantation therapy for acute myocardial infarction (AMI) in a miniswine model.</p><p><b>METHODS</b>CsA-NP emulsion was prepared by the high-pressure homogenization method. Models were performed by coronary angioplasty for percutaneous balloon occlusion of left anterior descending artery (LAD). A total of 17 miniswines survived after AMI were divided into four groups: control group (n=5), CsA-NP group (n=4), ASCs group (n=4), and CsA-NP+ASCs group (n=4). ASCs or saline were delivered by intracoronary injection one week after AMI.Before cell transplantation and 8 weeks after cell transplantation, delayed-enhanced magnetic resonance imaging (DE-MRI) was performed to evaluate cardiac function and viability. The infarcted myocardium and implanted cells were histologically studied.</p><p><b>RESULTS</b>Eight weeks after treatment, the left ventricular ejection fraction (LVEF)significantly increased in the CsA-NP+ASCs group when compared with the ASCs group [(53.6 ± 2.4)% vs. (48.3 ± 1.8)%, P<0.05]; meanwhile, the infarct size significantly decreased [(6.2 ± 1.7)cm(3) vs.(7.5 ± 0.6) cm(3), P<0.05] and the thickness of the ventricular wall significantly increased (P<0.05). Histology showed that the number of surviving cells increased nearly by three times in the CsA-NP+ASCs group, and the expressions of the cardiomyocyte specific markers (cTnT and α-actin) were detected. Histological samples also showed that CsA-NP+ASCs group reduced fibrotic tissue, and down-regulated the activation of Caspase-3.</p><p><b>CONCLUSION</b>The CsA-NP+ASCs combination therapy can enhance the viability of ASCs by improving LVEF and preventing LV expansion, which may be explained that CsA-NP has the anti-apoptotic effect and can promote the survivals and proliferation of ASCs.</p>