[Pretreatment with cyclosporin A nanoparticles emulsion protects apoptosis of swine adipose tissue-derived stem cells].

Objective: 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.

Methods: 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.

Results: 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.

Conclusions: 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.