Chen Huazhen, Inocencio Ryan, Alam Hasan B, Rhee Peter, Koustova Elena
Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.
J Surg Res. 2005 Feb;123(2):235-44. doi: 10.1016/j.jss.2004.08.025.
Matrix metalloproteinases (MMPs) and their specific physiological inhibitors, tissue inhibitors of metalloproteinases (TIMPs), are thought to play an essential role in tissue repair, cell death and morphogenesis. We have previously discovered unexpected up-regulation of genes coding for multiple MMP/TIMP family members in a rat model of hemorrhagic shock and resuscitation. However, the effect of different resuscitation protocols at the level of protein expression and function remains unknown.
Male Sprague-Dawley rats (n = 50; 10/group) were subjected to a three-stage volume controlled hemorrhage and resuscitated as follows: 1) lactated Ringer's solution (LR), 3:1 volume of lost blood; 2) 7.5% hypertonic saline (HTS), 9.7 ml/kg; 3) plasma, 1:1 volume. Sham hemorrhage and sham resuscitation groups were used as controls. Expression of lung and spleen MMPs (-2, -7, -9, -10, -14, and -16), and TIMPs (-1, -2, and -3) was analyzed at transcriptional, functional and protein expression level using RT-PCR, ELISA, Western blotting, and gelatin zymography techniques.
Spleen was affected more than lung by the resuscitation strategy and the largest number of changes was caused by HTS resuscitation. RT-PCR confirmed an increased levels of MMP-2, MMP-9, MMP-7, MMP-14, MMP-16, and TIMP-1, TIMP-2 in the spleen of HTS group compared to sham groups, whereas in lungs transcriptional levels of only TIMP-3 and TIMP-1 were significantly changed.
Expression of MMP and TIMP in lung and spleen following hemorrhage is modulated by the resuscitation strategy.
基质金属蛋白酶（MMPs）及其特异性生理抑制剂金属蛋白酶组织抑制剂（TIMPs）被认为在组织修复、细胞死亡和形态发生中起重要作用。我们之前在失血性休克和复苏的大鼠模型中发现了编码多个MMP/TIMP家族成员的基因意外上调。然而，不同复苏方案在蛋白质表达和功能水平上的影响仍然未知。
雄性Sprague-Dawley大鼠（n = 50；每组10只）经历三阶段容量控制性出血，并按以下方式进行复苏：1）乳酸林格氏液（LR），失血量的3倍体积；2）7.5%高渗盐水（HTS），9.7 ml/kg；3）血浆，1:1体积。假出血和假复苏组用作对照。使用逆转录聚合酶链反应（RT-PCR）、酶联免疫吸附测定（ELISA）、蛋白质印迹法和明胶酶谱技术，在转录、功能和蛋白质表达水平分析肺和脾中MMPs（-2、-7、-9、-10、-14和-16）以及TIMPs（-1、-2和-3）的表达。
复苏策略对脾的影响大于肺，且最大数量的变化是由HTS复苏引起的。RT-PCR证实，与假手术组相比，HTS组大鼠脾中MMP-2、MMP-9、MMP-7、MMP-14、MMP-16以及TIMP-1、TIMP-2的水平升高，而在肺中只有TIMP-3和TIMP-1的转录水平有显著变化。
出血后肺和脾中MMP和TIMP的表达受复苏策略调节。
