Research on mechanism of apoptosis and organ damage after hemorrhagic shock and resuscitation

将雄性BALB/c小鼠置于平均动脉压40+/-5 mmHg下90 min。通过4 ml/Kg的7.5%NaCl（高渗盐水; HS）和流出血（SB）或两倍流出血体积（2LR+SB）+SB或假手术（仅导管）或对照（无处理）进行复苏。（小肠）根据肠绒毛基底区空泡化程度设计肠损伤。采用末端脱氧核苷酸转移介导的5-三磷酸脱氧尿苷缺口末端标记法（TUNEL）和DNA梯状条带法检测小肠细胞凋亡评分。Western blotting检测caspase-3、HSP 70、HSP 40的表达。失血性休克和复苏后2 h的2LR+SB组细胞凋亡和6 h的2LR+SB组肠组织损伤较其他组明显（p<0.01），HS+SB 2 h组细胞凋亡指数较2LR+SB 2 h组明显降低（p<0.01），HS+ SB 6 h组肠组织损伤较2LR+SB 2 h组明显。 关于我们 与2LR+SB组相比，6 h时细胞凋亡率显著降低（p<0.01）。与2LR+SB 2 h组相比，HS+SB 2 h组caspase-3活性降低。HS+SB组2 h HSP 70、HSP 40表达高于2LR+SB组2 h。（肝、胸腺、脾）与2LR+SB组相比，HS+SB组2 h时肝、胸腺和脾的凋亡也减少（p<0.05）。HS+SB组2 h FAS和INK较2LR+SB组2 h略有降低。结论：高渗盐水复苏可减轻失血性休克复苏后肠组织细胞凋亡，减轻肠组织损伤。因此，高渗盐水复苏对细胞凋亡的影响可能在失血性休克复苏中起重要作用。HS复苏可能干扰小肠中的caspase-3活化。HS复苏可防止HSP 70和HSP 40的丢失，这可能与HS在小肠中的有益作用有关。FAS和INK可能参与了失血性休克复苏后肝脏的细胞凋亡。少

Male BALB/c mice were hemorrhaged to the mean arterial pressure of 40+/-5 mmHg for 90 min. Resuscitation was made by 4ml/Kg of 7.5% NaCl (hypertonic saline; HS) and shed blood (SB) or two times the volume of shed blood (2LR+SB) +SB or sham (catheter only) or control (no treatment).(Small intestine) Intestinal injury was devised and based on the extent of the vacuolation at basal area of the intestinal villi. Apoptosis score of the small intestines were examined by terminal deoxynucleotidyl transfer-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL) methods and DNA ladder assay. Activation of caspase-3, HSP 70, HSP 40 expression were assessed by Western blotting. Hemorrhagic shock and resuscitation induced the apoptosis in 2LR+SB at 2h and intestinal damage in 2LR+SB at 6h compared to other groups significantly (p<0.01) Apoptosis index of HS+SB at 2h group was significantly reduced compared to 2LR+SB at 2h (p<0.01) and also intestinal damage of HS+SB at 6h group was signiflc … More antly reduced compared to 2LR+SB at 6h (p<0.01). Activation of caspase-3 of HS+SB at 2h group was reduced compared to 2LR+SB at 2h group. On the contrary HSP 70, HSP 40 expression in HS+SB at 2h were higher than 2LR+SB at 2h.(Liver, Thymus, Spleen) Apoptosis of liver, thymus and spleen were also reduced in HS+SB at 2h compared to 2LR+SB at 2h (p<0.05). FAS and INK in HS+SB at 2h were slightly reduced than 2LR+SB at 2h.Conclusions : Hypertonic saline resuscitation reduced the apoptosis and intestinal damage after hemorrhagic shock and resuscitation. Thus, the effect of hypertonic saline resuscitation toward apoptosis may have important role in hemorrhagic shock and resuscitation. HS resuscitation may interfere with caspase-3 activation in the small intestine. HS resuscitation may prevent the loss of HSP 70 and HSP 40, which might be involved in the beneficial effect for HS in the small intestine. FAS and INK may be involved in the apoptosis of the liver after hemorrhagic shock and resuscitation. Less