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

失血性休克复苏后细胞凋亡及器官损伤机制研究

• 批准号: 14571461
• 负责人: MURAO Yoshinori
• 金额: $ 2.24万
• 依托单位: Nara Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571461/
• 关键词: Hemorrhagic Shock; Hypertonic Saline; Apoptosis; TUNEL; DNA ladder; Caspase-3; HSP 70; HSP 40; JNK; FAS; Caspase-3; HSP 40; HSP 70

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

雄性 BALB/c 小鼠失血至平均动脉压 40+/-5 mmHg，持续 90 分钟。用4ml/Kg 7.5% NaCl（高渗盐水；HS）和流血（SB）或两倍流血量（2LR+SB）+SB或假手术（仅导管）或对照（无处理）进行复苏。（小肠）根据肠绒毛基底区域空泡化的程度设计肠损伤。通过末端脱氧核苷酸转移介导的脱氧尿苷5-三磷酸缺口末端标记(TUNEL)方法和DNA梯测定法检查小肠细胞凋亡评分。通过蛋白质印迹评估 caspase-3、HSP 70、HSP 40 表达的激活。失血性休克和复苏诱导 2LR+SB 2h 细胞凋亡和 2LR+SB 6h 肠道损伤与其他组相比显着 (p<0.01) HS+SB 2h 组细胞凋亡指数较 2LR+SB 2h 显着降低 (p<0.01) 并且 HS+SB 6h 组肠道损伤显着 ... 更多 6小时时与2LR+SB相比显着降低(p<0.01)。与2LR+SB 2h组相比，2h组HS+SB的caspase-3活化降低。相反，HS+SB 2h 中的 HSP 70、HSP 40 表达高于 2LR+SB 2h 中的表达。(肝脏、胸腺、脾脏) 与 2LR+SB 2h 相比，HS+SB 2h 中肝脏、胸腺和脾脏的凋亡也减少(p<0.05)。 HS+SB 2h 时的FAS 和INK 较2LR+SB 2h 时略有减少。结论：高渗盐水复苏可减轻失血性休克复苏后的细胞凋亡和肠道损伤。因此，高渗盐水复苏对细胞凋亡的影响可能在失血性休克和复苏中具有重要作用。 HS 复苏可能会干扰小肠中 caspase-3 的激活。 HS 复苏可以防止 HSP 70 和 HSP 40 的损失，这可能与小肠中 HS 的有益作用有关。 FAS和INK可能参与失血性休克和复苏后肝脏的凋亡。较少的

项目成果

Murao Y, Loomis W, Wolf P, Hoyt DB, Junger WG: "Effect of dose of hypertonic saline on its potential to prevent lung tissue damage in a mouse model of hemorrhagic shock"Shock. 20. 29-34 (2003)

Murao Y、Loomis W、Wolf P、Hoyt DB、Junger WG：“高渗盐水剂量对其预防失血性休克小鼠模型肺组织损伤的潜力的影响”休克。

Mura Y, Hata M, Ohnishi K, Okuchi K, Nakajima Y, hiasa Y, Junger WG, Hoyt DB, Ohnishi T: "Hypertonic Saline Resuscitation Reduces Apoptosis and Tissue Damage of the Small Intestine in a Mouse Model of Hemorrhagic Shock"Shock. 20. 23-28 (2003)

Mura Y、Hata M、Ohnishi K、Okuchi K、Nakajima Y、hiasa Y、Junger WG、Hoyt DB、Ohnishi T：“高渗盐水复苏可减少失血性休克小鼠模型中小肠的细胞凋亡和组织损伤”休克。

Murao Y, Hata M, Ohnish K, Okuchi K, Nakajima Y et al.: "Hypertonic saline resuscitation reduces apoptosis and tissue damage of the small intestine in a mouse model of hemorrhagic shock"Shock. 20. 23-28 (2003)

Murao Y、Hata M、Ohnish K、Okuchi K、Nakajima Y 等人：“高渗盐水复苏可减少失血性休克小鼠模型中小肠的细胞凋亡和组织损伤”。

Murao Y, Hata M, Ohnishi K, Okuchi K, Nakajima Y, Hiasa Y, Junger WG, Hoyt DB, Ohnishi T: "Hypertonic saline resusitaion reduces apoptosis and tissue damage of the small intestine in a mouse model of hemorrhagic shock"Shock. 20. 23-28 (2003)

Murao Y、Hata M、Ohnishi K、Okuchi K、Nakajima Y、Hiasa Y、Junger WG、Hoyt DB、Ohnishi T：“高渗盐水复苏可减少失血性休克小鼠模型中小肠的细胞凋亡和组织损伤”休克。

Murao Y, Loomis W, Wolf P, Hoyt DB, Junger WG: "Effect of Dose of Hypertonic Saline on its Potential to Prevent Lung Tissue Damage in a Mouse Model Hemorrhagic Shock"Shock. 20. 29-34 (2003)

Murao Y、Loomis W、Wolf P、Hoyt DB、Junger WG：“高渗盐水剂量对其预防失血性休克小鼠模型肺组织损伤的潜力的影响”休克。