A Novel Preservation Solution Prolonged Cold Storage in Mouse Cardiac Grafts.

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Background: We developed a novel non-potassium preservation solution contains L-cysteine, glycine, ascorbic acid/ascorbic acid-2-phosphate magnesium to prolonged mouse cardiac grafts cold storage. Methods: B6 mice underwent heterotopic heart transplantation and the animals were derived into 3 groups: recipients with non-preserved grafts (control group), recipients with grafts preserved in HTK solution for 24 and 48 hours (HTK-group), and recipients with grafts preserved in the new solution, called Solution II for 24 and 48 hours (S-II group). Recipient serum and grafts were harvest for assay. Results: In both 24hr- and 48hr-preservation series (24h- and 48h-series), time for restoration of heartbeat after reperfusion in S-II group was significantly shorter than in HTK group (HTK-24h: 306.2±39.6sec, S-II-24h: 110.5±20.3sec, p<0.01; HTK-48h: 1537.5±454.7sec, S-II-48h: 640±286sec, p<0.01). In 48h-series, Release of serum LDH, CK in S-II-group was significantly lower to compare with HTK group. The graft survival of HTK-48h group is 0 day (n=9), but S-II group is 9d, 17d, >100d×2. Graft ATP level of S-II-48h was significantly higher than HTK-48h. In both 24 and 48h-series, S-II significantly inhibited graft apoptosis. Moreover, immunohistochemistry staining revealed that the CD68-positive macrophage infiltration in S-II group was significantly lower compared with HTK group. In 24h-series, graft 8-OHdG positive cells (%) were significantly lower in S-II compared with HTK. Consistent with histologic findings, serum 8-OHdG levels were significantly inhibited in S-II group. Furthermore, the mRNA expression of the IL-6, HO-1, iNOS, CD11b in S-II-48h group, were significantly lower, wherever Nrf2, NF-κB, Arg-1, HIF-1α, TGF-β were significantly higher than HTK-48h group (p<0.01). Conclusion: Hence our data indicated that the novel preservation solution impairs graft macrophage infiltration, inhibits inflammatory cytokines and myocardial apoptosis which results in attenuating mouse cardiac grafts cold ischemia-reperfusion injury and prolonging cold storage.

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