摘要
目的 观察失血性休克后不同时期大鼠肠系膜上动脉平滑肌FK506结合蛋白12.6（FKBP12.6）的表达及其分布,检测休克不同时相大鼠肠系膜上动脉血管反应性的变化,探讨FKBP12.6是否参与对失血性休克血管反应性的调节.方法 建立大鼠失血性休克[40 mm Hg（1 mm Hg =0.133 kPa）]早期（30 min）及晚期（2 h）模型；采用离体器官张力测定技术检测失血性休克后不同时相大鼠肠系膜上动脉主干对缩血管物质去甲肾上腺素（NE）的收缩反应性；采用Western blot法检测失血性休克不同时相大鼠肠系膜上动脉平滑肌FKBP12.6的表达及其分布；采用免疫共沉淀联合免疫杂交检测FKBP12.6-雷诺定敏感受体2（RyR2）复合子形成.结果 失血性休克早期（30 min）,大鼠肠系膜上动脉对NE的收缩反应性明显提高,最大收缩力（Emax）由（0.70 ±0.18）g升至（1.03±0.31）g（P ＜0.05）,而在休克晚期（2 h）大鼠肠系膜上动脉对NE的收缩反应性明显降低,Emax由（0.70 ±0.18）g降至（0.54±0.12） g（P＜0.05）,提示在失血性休克不同时相,大鼠肠系膜上动脉对NE具有“双相”反应性；失血性休克后30 min及2h,大鼠肠系膜上动脉血管平滑肌FKBP12.6总蛋白的表达无明显变化,但在失血性休克早期,FKBP12.6在胞质内的表达明显增强,在质膜上的表达明显降低；在休克晚期,FKBP12.6在胞质内的表达降低而在质膜的表达升高；免疫共沉淀结果可见,在失血性休克30 min,大鼠肠系膜上动脉平滑肌FKBP12.6-RyR2复合子的形成明显减少,而在休克晚期FKBP12.6-RyR2复合子的形成较正常对照比较无明显改变；相关分析显示,失血性休克早期FKBP12.6在胞质内的表达增强与休克早期血管高反应性呈显著正相关（r =0.993,P＜0.01）.结论 在失血性休克不同时相,大鼠肠系膜上动脉平滑肌FKBP12.6存在胞质/质膜的转位,在休克早期FKBP12.6从RyR2的解离,至少部分促成失血性休克早期大鼠肠系膜上动脉对NE高反应性的形成.
Objective To explore the expression and distribution of FKS06 binding protein 12. 6 （FKBP12. 6） in superior mesenteric artery （SMA） during different stages after hemorrhagic shock and its role in the regulation of vascular reactivity during different stages after hemorrhagic shock. Methods The early （30 rain） and late stage （2 h） after hemorrhagic shock [40 mm Hg（ 1 mm Hg =0. 133 kPa） ] mod els were established in rats. The vascular reactivity of SMA to norepinephrine （NE） was measured in organ perfusion chamber. The expression and distribution of FKBP12. 6 at different stages after hemorrhagic shock were detected by using Western blotting. The formation of FKBP12. 6-ryanodine sensitive receptor 2 （RyR2） complex was measured with co-immunoprecipitation （IP） and Western blotting. Results During early stage （30 min） after hemorrhagic shock, the vascular reactivity to NE was increased significantly, and Emx increased from （ 0. 70 ± 0. 18 ） g to （ 1.03 ± 0. 31 ） g （ P 〈 0. 05 ）. The vascular reactivity to NE during late stage （2 h） was decreased, and Emx decreased from （0.70 ±0. 18 ） g to （0. 54 ± 0. 12） g,which suggested that there was bi-phasic vascular reactivity to NE in SMA during different stages after hem orrhagic shock. In addition, the total expression of FKBP12. 6 had no significant changes but the distribu- tion of FKBP12. 6 in cytoplasm and plasmalemma changed significantly, characterized with the expression of FKBP12. 6 during early stage （30 min） after hemorrhagic shock in cytoplasm was upregulated while de creased in plasmalemma significantly. During late stage （2 h） after hemorrhagic shock, the expression of FKBP12. 6 in cytoplasm was downregulated but its expression in plasmalemma increased significantly no matter compared with control or shock 30 rain group. Co-IP analysis results showed that formation of FKBP12. 6-RyR2 complex was decreased during early stage after hemorrhagic shock, while there was no significant changes of the FKBP12. 6-RyR2 complex formation during late stage after hemorrhagic shock. The correlation analysis showed that the increased expression of FKBP12. 6 in cytoplasm during early stage after hemorrhagic shock was closely associated with the development of vascular hyperreactivity to NE after hemorrhagic shock （ r = 0. 993,P 〈 0. 01 ）. Conclusion At different stages after hemorrhagic shock, there were the translocations of FKBP12. 6 from cytoplasm to plasmalemma in SMA in rats. At early stage after hemorrhagic shock, the dissociation of FKBP12. 6 from RyR2 was at least partly associated with the development of vascular hvoerreaetivitv to NE.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第11期2284-2288,共5页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81100227、81370427）
