重度烫伤小鼠血清及去补体后血清诱导巨噬细胞凋亡作用的体外实验研究

In vitro study of macrophages apoptosis induced by serum and decomplemented serum from severely scalded mice

目的 探讨重度烫伤小鼠血清及去补体血清在体外诱导腹腔巨噬细胞 (pMФs)凋亡及其机制。方法 采用小鼠 30 %TBSAⅢ度烫伤模型 ,分为正常对照组、烫伤组、去补体烫伤组 ,检测各组伤后 6h血清对体外培养的pMФs分泌超氧阴离子 (O2- )和一氧化氮 (NO)产量的影响 ,碘化丙锭 (PI)染色流式细胞术及凋亡电泳试验测定pMФs的凋亡情况。结果 与正常对照测值比较 ,烫伤血清诱导pMФs分泌较多的O2- 和NO ,去补体后烫伤血清诱导O2- 和NO的分泌量显著降低。烫伤血清诱导pMФs凋亡明显增加 ,去补体后烫伤血清和活性氧阻断剂PDTC及NO阻断剂AG能阻断绝大部分pMФs的凋亡。结论 重度烫伤小鼠血清补体能在体外诱导pMФs凋亡 ,O2-

Objective To study apoptotic changes and mechanism of cultured peritoneal macrophages (pMФs) in response to the serum and decomplemented serum from severely burned mice. Methods The mice were randomly divided into normal control, scald group, and decomplemented scald group. Mice in scald and decomlemented scald groups were scalded to 30% total body surface area (TBSA) Ⅲ degree. The production of superoxide anion (O 2 -) and nitric oxide (NO) by peritoneal macrophages from normal mice were incubated with serum from every group was examined. The apoptosis percentage of the pMФs was measured by flow cytometry and propidium iodide staining, and the apoptotic fragmentation at genome DNA of pMФs was detected by electrophoresis assay. Results Serum from injured mice could induce pMФs to produce more O 2 - and NO. The percentage of apoptotic macrophages induced by the serum from burned mice was significantly increased. The serum after decomplemented burned mice, PDTC (Inhibitor of reactive oxygen species), and AG (Inhibitor iNOS) could block the apoptosis of pMФs. Conclusin Macrophage apoptosis can be induced by serum complement from scalded mice. These evidences suggested that O 2 - and NO may be involved in the apoptosis of pMФs induced by the serum complement from scalded mice.