Cotogni Paolo, Bini Roberto, Trombetta Antonella, Olivero Giorgio
Anestesiologia e Rianimazione, Dipartimento di Discipline Medico-Chirurgiche, Università di Torino, Torino, Italia.
J Invest Surg. 2010 Dec;23(6):295-302. doi: 10.3109/08941939.2010.497239.
The aim of this study is to evaluate whether hemorrhage and resuscitation affect liver, intestinal, and renal expressions of heat shock protein 70 (HSP70), inducible nitric oxide synthase (iNOS), and apoptosis indexes (TUNEL, caspase-3 activation) and whether the expression of these proteins can be modulated by pyrrolidine dithiocarbamate (PDTC) after a nonlethal hemorrhagic shock (HS) in rats.
Forty rats were randomized into four groups: sham-operated, only HS, HS/resuscitation with blood plus normal saline (NS), and HS/resuscitation with blood/NS plus PDTC, 15 mg/kg body weight, intravenously. Rats were subjected to HS by blood removal to a mean arterial pressure of 35-40 mmHg through the femoral artery. After 1 hr of shock, the animals were resuscitated according to the experimental protocol. HSP70, iNOS, cleaved caspase-3 expression, and TUNEL were analyzed in liver, small intestine, and kidney 3 hr after resuscitation.
HS upregulated HSP70, iNOS, cleaved caspase-3 expression, and induced apoptosis (TUNEL) (p < .05 to < .001). Resuscitation was not associated with further increase of their expressions. The administration of PDTC during resuscitation decreased liver, intestinal, and renal activation of iNOS and apoptosis indexes (p < .05 to < .001), and was associated with further increase in HSP70 expression (p < .05).
Our results show that HS resuscitation with PDTC modulates several signaling pathways (HSP70, iNOS, TUNEL, and caspase-3) in a rat model. The results suggest that PDTC administration--by reducing apoptosis and iNOS expression--may have a potential role in minimizing organ damage after severe hemorrhage.
本研究旨在评估出血和复苏是否会影响热休克蛋白70（HSP70）、诱导型一氧化氮合酶（iNOS）在肝脏、肠道和肾脏中的表达以及凋亡指标（TUNEL、半胱天冬酶-3激活），并评估在大鼠非致死性失血性休克（HS）后，这些蛋白的表达是否能被吡咯烷二硫代氨基甲酸盐（PDTC）调节。
40只大鼠随机分为四组：假手术组、单纯HS组、用血液加生理盐水（NS）进行HS/复苏组、用血液/NS加15mg/kg体重的PDTC静脉注射进行HS/复苏组。通过股动脉放血使大鼠平均动脉压降至35 - 40mmHg来诱导HS。休克1小时后，根据实验方案对动物进行复苏。复苏3小时后，分析肝脏、小肠和肾脏中HSP70、iNOS、裂解的半胱天冬酶-3表达以及TUNEL情况。
HS上调了HSP70、iNOS、裂解的半胱天冬酶-3表达并诱导了凋亡（TUNEL）（p <.05至<.001）。复苏并未使其表达进一步增加。复苏期间给予PDTC可降低肝脏、肠道和肾脏中iNOS的激活及凋亡指标（p <.05至<.001），并与HSP70表达的进一步增加相关（p <.05）。
我们的结果表明，在大鼠模型中，用PDTC进行HS复苏可调节多种信号通路（HSP70、iNOS、TUNEL和半胱天冬酶-3）。结果表明，给予PDTC通过减少凋亡和iNOS表达，可能在减轻严重出血后的器官损伤方面具有潜在作用。
