Extracellular splitting pattern of mitochondria and the depressant effects of CsA on the process / 中国骨伤

ABSTRACT

<p><b>OBJECTIVE</b>To investigate extracellular splitting pattern of mitochondria and the depressant effects of CsA on the process and explore the mechanism of post-traumatic SIRS and its therapeutic strategy.</p><p><b>METHODS</b>Ten male SD rats with 60 to 70 days age and 240 to 280 g weight were used for mitochondrial isolation. Freshly isolated mitochondria were randomly divided into two groups, which were cultured in blood plasma with or without CsA respectively for 8 h. COX and MDH were assayed by ELISA every 30 min. Meanwhile, Rat macrophage cell line NR8383 were treated as follows, control (group A) cultivation with normal medium; NR8383+CsA co-culture group (group B) culture medium was supplemented with CsA of 10 mmol/L; NR8383+intact mitochondria co-culture group (group C) culture medium was supplemented with intact mitochondria (mtDNA=5 g/ml); NR8383+intact mitochondria+CsA co-culture group (group D) culture medium was supplemented with intact mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L; NR8383+disrupted mitochondria co-culture group (group E) culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml); NR8383+disrupted mitochondria+CsA co-culture group (group F) culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L. TNF-α and IL-6 concentrations in supernatant were assessed at 1, 3, 5 h after culture.</p><p><b>RESULTS</b>In the mitochondria plasma cultures, MDH and COX levels were increased with the time and peaked at about 3 h and 3.5 h; CsA can delay the appearance of peak to 4.5 h. Among different treated groups,there was no significant difference in TNF-α and IL-6 between group A and group B; there was significant difference in TNF-α and IL-6 other groups. After 1 h culture, compared with group C, no significant difference of TNF-α and IL-6 was observed in group D, while TNF-α and IL-6 were significant higher in group E; after 3 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while TNF-α and IL-6 were significantly higher in group E; after 5 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while no significant difference of TNF-α and IL-6 were observed in group E. At each time point, there was no significant difference in TNF-α and IL-6 between group F and group E.</p><p><b>CONCLUSION</b>Mitochondria can split in serum with time, which will further activate macrophages. CsA has depressant effect to mitochondrial splitting on the process and will therefore inhibit the activation of macrophages.</p>